compound, pharmaceutical composition, and, method of making a compound

专利摘要:
COMPOUND, PHARMACEUTICAL COMPOSITION AND METHOD OF MAKING A COMPOUND Compounds of formula I, compositions containing them, and methods of use for the compounds and with positions in the treatment of conditions in which modulation of the JAK pathway or inhibition of JAK kinases are disclosed , particularly JAK 2 and JAK3, are therapeutically useful. Methods of making the compounds are also disclosed.
公开号:BR112013001632B1
申请号:R112013001632-9
申请日:2011-07-27
公开日:2021-05-25
发明作者:Hui Li；Thilo J. Heckrodt；Yan Chen；Darren John Mcmurtrie；Vanessa Taylor；Rajinder Singh；Pingyu Ding；Rose Yen
申请人:Rigel Pharmaceuticals；
IPC主号:

专利说明:

INTRODUCTION Field
[0001] The present disclosure relates to compounds and methods for their use in modulating the JAK pathway, inhibiting one or more JAK kinases and treating conditions in which modulating the JAK pathway or inhibiting JAK kinases, particularly JAK3 are therapeutically useful. Fundamentals
Janus Kinases (or JAK) are a family of cytoplasmic protein tyrosine kinases that includes JAK1, JAK2, JAK3 and TYK2. Each of the JAK kinases is selective for receptors for certain cytokines, although multiple JAK kinases can be affected by particular cytokine or signaling pathways. Studies suggest that JAK3 associates with the common gamma chain (70) of various cytokine receptors. In particular, JAK3 selectively binds to receptors and is part of the cytokine signaling pathway for IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21. The JAK1 kinase interacts, among others, with the receptors for the cytokines IL-2, IL-4, IL-7, IL-9 and IL-21, while the JAK2 interacts, among others, with the receptors for IL-9 and TNF-α. Upon binding of certain cytokines to their receptors (eg, IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21), receptor oligomerization occurs, resulting in the cytoplasmic tails of JAK kinases associated being brought in close proximity and facilitating the trans-phosphorylation of tyrosine residues in the JAK kinase. This trans-phosphorylation results in activation of the JAK kinase.
[0003] Phosphorylated JAK kinases bind to various Signal Transducer and Transcription Activator (STAT) proteins. These STAT proteins, which are DNA-binding proteins activated by phosphorylation of tyrosine residues, function as both signaling molecules and transcription factors and ultimately bind to specific DNA sequences present in cytokine-responsive gene promoters (Leonard et al., (2000), J. Allergy Clin. Immunol. 105: 877-888). JAK/STAT signaling has been implicated in mediating many abnormal immune responses such as allergies, asthma, autoimmune diseases such as transplant rejection (allograft), rheumatoid arthritis, amyotrophic lateral sclerosis and multiple sclerosis, as well as solid and hematological malignancies such as like leukemia and lymphomas. For a review of the pharmaceutical intervention of the JAK/STAT pathway see Frank, (1999), Mol. Med. 5: 432: 456 and Seidel et al., (2000), Oncogene 19: 2645-2656.
[0004] In particular, JAK3 has been implicated in a variety of biological processes. For example, IL-4 and IL-9 induced proliferation and survival of murine mastoids have been shown to be dependent on JAK3 and gamma chain signaling ( Suzuki et al., (2000), Blood 96: 2172-2180 ). Playing a crucial role in IgE receptor-mediated mastoid degranulation responses (Malaviya et al., (1999), Biochem. Biophys. Res. Commun. 257: 807-813), inhibition of JAK3 kinase has been shown to prevent JAK3 kinase reactions. type I hypersensitivity, including anaphylaxis ( Malaviya et al., (1999), J. Biol. Chem. 274:27028-27038 ). Inhibition of JAK3 has also been shown to result in immune suppression for allograft rejection (Kirken, (2001), Transpl. Proc. 33: 3268-3270). Kinases, particularly JAK3 kinases, have also been implicated in the mechanism involved in early and later stages of rheumatoid arthritis (Muller-Ladner et al., (2000), J. Immunol. 164: 3894-3901); familial amyotrophic lateral sclerosis ( Trieu et al., (2000), Biochem Biophys. Res. Commun. 267: 22-25 ); leukemia ( Sudbeck et al., (1999), Clin. Cancer Res. 5: 1569-1582 ); fungoid mycosis, a form of T-cell lymphoma (Nielsen et al., (1997), Prac. Natl. Acad. Sci. USA 94: 6764-6769); and abnormal cell growth ( Yu et al., (1997), J. Immunol. 159: 5206-5210; Catlett-Falcona et al., (1999), Immunity 10: 105-115).
[0005] JAK kinases, including JAK3, are abundantly expressed in primary leukemic cells of children with acute lymphoblastic leukemia, the most common form of childhood cancer, and studies have correlated STAT activation in certain cells with signals that regulate apoptosis ( Demoulin et al., (1996), Mol.Cell.Biol. 16:4710-6; Jurlander et al., (1997), Blood.89:4146-52; Kaneko et al., (1997), Clin.Exp Immun. 109: 185-193; and Nakamura et al., (1996), J. Biol. Chem. 271: 19483-8). They are also known to be important for lymphocyte differentiation, function and survival. In particular, JAK3 plays an essential role in the function of lymphocytes, macrophages, and mastoids. Given the importance of JAK kinases, particularly JAK3, compounds that modulate the JAK pathway, including those selective for JAK3, may be useful to treat diseases or conditions where lymphocyte, macrophage, or mastoid function is involved (Kudlacz et al. , (2004) Am. J. Transplant 4: 51-57; Changelian (2003) Science 302: 875-878). Conditions in which JAK pathway targeting or modulation of JAK kinases, particularly JAK3, are considered to be therapeutically useful include, leukemia, lymphoma, transplant rejection (eg pancreatic islet transplant rejection, bone marrow transplant applications (eg, graft-versus-host disease), autoimmune diseases (eg, diabetes), and inflammation (eg, asthma, allergic reactions) Conditions that may benefit from JAK3 inhibition are discussed in more detail below .
[0006] In view of the numerous conditions that are considered to benefit from treatment involving modulation of the JAK pathway it is immediately evident that new compounds that modulate JAK pathways and methods of using these compounds should provide substantial therapeutic benefits to a wide range of patients. Provided herein are novel 2,4-pyrimidinediamine compounds for use in treating conditions where targeting the JAK pathway or inhibiting JAK kinases, particularly JAK3, is therapeutically useful. SUMMARY
[0007] In one embodiment, the present disclosure is directed to compounds, prodrugs, and methods of using these compounds and their prodrugs in treating conditions in which modulation of the JAK pathway or inhibition of JAK kinases , particularly JAK2, JAK3, or both will be therapeutically useful.
[0008] One embodiment is a compound of formula I, a salt thereof, or a pharmaceutical composition which includes the compound:

[0009] X and Y are each independently O, S, S(O), SO2 or NR1;
[00010] each R1 is independently for each occurrence H, optionally substituted C1-6 alkyl, C(O)-C1-6 alkyl, CO2-C1-6 alkyl or R50;
[00011] each R50 is -C(R9)2-A-R10, where A is O or S; each R9 is independently for each occurrence H, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl, or optionally substituted C7-16 arylalkyl; or alternatively, two R 9 , together with the carbon to which they are attached, form an optionally substituted C 3-8 cycloalkyl group or an optionally substituted 3- to 8-membered heteroarylcyclyl group; R10 is Ra, -P(O)(OR11)2, -P(O)(OR11)N(R12)2 or -P(O)(N(R12)2)2; each R11 is independently for each occurrence Ra or a monovalent cationic group; either two R11 together with the atoms to which they are attached form a 4- to 8-membered cyclic phosphate group, or two R11 together represent a divalent cationic group; each R12 is independently for each occurrence Rc or -C1-3 alkyl -N(Rc)2; or two R12, each on nitrogens separate from -P(O)(N(R12)2)2, together with the atoms to which they are attached, form a 4- to 8-membered cyclic phosphonic acid bisamide group; or an R12 together with R11 of the group -P(O)(OR11)N(R12)2, together with the atoms to which they are attached, form a 4- to 8-membered cyclic phosphonamidate group;
[00012] ring A is a C6-10 aryl or a 5- to 10-membered heteroaryl; each R2 is independently for each occurrence H, Re, Rb, Re substituted with one or more of the same or different Ra, Rb or both, -ORe substituted with one or more of the same or different Ra and/or Rb, -SRe substituted with one or more of the same or different Ra and/or Rb, -C(O)Re substituted with one or more of the same or different Ra and/or Rb, -N(Ra)Re where Re is substituted with one or more of the same or different Ra and/or Rb, - S(O)2Re substituted with one or more of the same or different Ra and/or Rb, - N(Ra)-S(O)2Re where Re is substituted with one or more of the same or different Ra and/or Rb, -B(ORa)2, -B(N(Rc)2)2, -(C(Ra)2)m-Rb, -O-(C(Ra)2)m- Rb, -abba ab S-(C(R)2)mR, -O-(C(R)2)mR, -N(R)-(C(R)2)mR, -O-(CH2 )m- bb aabaa CH((CH2)mR )R , -C(O)N(R )-(C(R )2)mR , -O-(C(R )2)mC(O)N(R ) )- ab ab aaab (C(R )2)mR , -N((C(R )2)mR )2, -S-(C(R )2)mC(O)N(R )-(C( R )2)mR , - aaabaa ab N(R )-C(O)-N(R )-(C(R )2)mR , -N(R )-C(O)-(C(R )2 )mC(R )(R )2 or - N(Ra)-(C(Ra)2)mC(O)-N(Ra)- ( C(Ra)2)m-Rb;
[00013] each Ra is independently for each occurrence H, deuterium, C1-6 alkyl, C3-8 cycloalkyl, C4-11 cycloalkylalkyl, C6-10 aryl, C716 arylalkyl, 2 to 6 membered heteroalkyl, 3 to 10 membered heteroalicyclyl , 4- to 11-membered heteroalicyclylalkyl, 5- to 15-membered heteroaryl, or 6- to 16-membered heteroarylalkyl;
[00014] each Rb is independently for each occurrence =O, -ORa, -O-(C(Ra)2)m-ORa, halo-C1-3 alkyloxy, =S, -SRa, =NRa, =NORa, - N(Rc)2, halo, -CF3, -CN, -NC, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)Ra, -S(O)2Ra, - SO3Ra, -S(O)N(Rc)2, -S(O)2N(Rc)2, -OS(O)Ra, -OS(O)2Ra, -OSO3Ra, - OS(O)2N(Rc) 2, -C(O)Ra, -CO2Ra, -C(O)N(Rc)2, -C(NRa)-N(Rc)2, -C(NOH)-Ra, -C(NOH)-N (Rc)2, -OC(O)Ra, -OC(O)ORa, -OC(O)N(Rc)2, -OC(NH)-N(Rc)2, -ac aaaaa OC(NR )- N(R )2, -N(R )-S(O)2H, -[N(R )C(O)]nR , -[N(R )C(O)]nOR , - [N(Ra) C(O)]nN(Rc)2 or -[N(Ra)C(NRa)]nN(Rc)2;
[00015] each Rc is independently for each occurrence Ra, or, alternatively, two Rc are taken together with the nitrogen atom to which they are attached to form a 3- to 10-membered heteroallylcyclyl or a 5- to 10-membered heteroaryl that can optionally including one or more of the same or different additional heteroatoms and which are optionally substituted with one or more of the same or different Ra and/or Rd groups;
[00016] each Rd is =O, -ORa, halo-C1-3 alkyloxy, C1-6 alkyl, =S, -SRa, =NRa, =NORa, -N(Ra)2, halo, -CF3, -CN , -NC, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)Ra, -S(O2)Ra, -SO3Ra, -S(O)N(Ra)2, -S(O)2N(Ra)2, -OS(O)Ra, - OS(O)2Ra, -OSO3Ra, -OS(O)2N(Ra)2, -C(O)Ra, -CO2Ra, - C(O)N(Ra)2, -C(NRa)N(Ra)2, -C(NOH)Ra, -C(NOH)N(Ra)2, -OCO2Ra, -OC(O)N(Ra )2, - aa aaaaaa OC(NR )N(R )2, -[N(R )C(O)]nR , -(C(R )2)n-OR , -N(R )-S(O )2R , -C(O)-C1-6 haloalkyl, -S(O)2 C1-6 haloalkyl, -OC(O)Ra, -O(C(Ra)2)m -ORa, -a aa a aa a S(C(R )2)m-OR , -N(R ) C1-6 haloalkyl, -P(O)(OR )2, -N(R )-(C(R )2)m -OR , -[N(Ra)C(O)]nORa, -[N(Ra)C(O)]n N(Ra)2, -[N(Ra)C(NRa)]nN(Ra)2 or -N (Ra)C(O) haloC 1-6 alkyl; two Rd, taken together with the atom or atoms to which they are attached, combine to form a partially or fully saturated 3- to 10-membered mono or bicyclic ring, optionally containing one or more heteroatoms and optionally substituted with one or more Frog;
[00017] each Re is independently for each occurrence C1-6 alkyl, C3-8 cycloalkyl, C4-11 cycloalkylalkyl, C6-10 aryl, C7-16 arylalkyl, 2 to 6 membered heteroalkyl, 3 to 10 membered heteroalicyclyl, heteroalicyclylalkyl 4- to 11-membered, 5- to 15-membered heteroaryl, or 6- to 16-membered heteroarylalkyl;
[00018] p is 0, 1, 2, 3 or 4;
[00019] each m is 1, 2 or 3;
[00020] each n is 0, 1, 2 or 3;
[00021] two R2 groups, taken together with the atom or atoms to which they are attached, combine to form a partially or fully saturated 4- to 10-membered mono or bicyclic ring, optionally containing one or more heteroatoms and optionally substituted with one or more Ra and/or Rb;
[00022] Z1 and Z2 are each independently CH, CR2 or N;
[00023] R3 is H, optionally substituted C1-6 alkyl or R50;
[00024] R4 is H, optionally substituted C1-6 alkyl or R50; and
[00025] R5 is H, halo, -CN, optionally substituted C1-6 alkyl, alkynyl, hydroxy, optionally substituted C1-6 alkoxy, nitro, -N(Ra)2, -C(O)N(Ra)2, -CO2Ra or -C(O)Ra.
[00026] Another embodiment is a method of inhibiting a JAK kinase activity, which includes contacting the JAK kinase with an amount of a compound effective to inhibit a JAK kinase activity where the compound conforms to formula I as herein. described. In one embodiment the contact is made in vitro, in another embodiment the contact is made in vivo.
[00027] Another embodiment is a method of treating a T-cell mediated autoimmune disease, which includes administering to a patient suffering from such an autoimmune disease an amount of a compound effective to treat the autoimmune disease where the compound is in accordance. with formula I as described herein.
[00028] Another embodiment is a method of treating allograft transplant rejection in a transplant recipient, which includes administering to the transplant recipient an amount of a compound effective to treat or prevent rejection where the compound is in accordance with formula I as described herein. Administration in this context may include contacting a transplanted organ with a compound or pharmaceutical composition described herein prior to transplantation and/or concurrent with administration to the transplant recipient.
[00029] Yet another embodiment is a method of treating a Type IV hypersensitivity reaction, which includes administering to a patient an amount of a compound effective to treat or prevent the hypersensitivity reaction where the compound conforms to formula I as described here.
[00030] Another embodiment is a method of treating an ocular disease or disorder, which includes administering to a patient an amount of a compound effective to treat or prevent the ocular disease or disorder where the compound is in accordance with formula I as described here.
[00031] Another embodiment is a method of inhibiting a signal transduction cascade in which JAK3 kinase plays a role, which includes contacting a cell expressing a receptor involved in such a signaling cascade with a compound where the compound is according to formula I as described herein.
[00032] One embodiment is a method of treating a disease mediated by JAK kinase, which includes administering to a patient an amount compound effective to treat or prevent the disease mediated by JAK kinase where the compound is in accordance with formula I as described here.
[00033] Another embodiment is a pharmaceutical formulation which includes a compound of formula I as described herein. Therapy using the 2,4-pyrimidinediamine compounds and pharmaceutical formulations described herein can be applied alone, or it can be applied in combination with or adjunct to other immunosuppressive therapies.
[00034] Another embodiment is a kit which includes a compound of formula I as described herein, a prodrug thereof or pharmaceutical composition which includes a compound thereof, packaging and instructions for use.
[00035] Another embodiment is a single dosage formulation which includes a compound of formula I as described herein, a prodrug thereof or pharmaceutical composition which includes a compound of formula I.
[00036] Other embodiments include methods of using the compound to screen for other agents used to treat or prevent a disease mediated by JAK kinase.
[00037] Other embodiments include methods of making the compounds described herein.
[00038] More detailed description for these and other embodiments is provided below. DETAILED DESCRIPTION Overview
[00039] The invention includes the compounds of formula I and compositions and methods using these compounds in the treatment of conditions where modulation of the JAK pathway or inhibition of JAK kinases, particularly JAK3, are therapeutically useful. The formulations, uses as screening agents and other utilities are also described. Terms
[00040] As used herein, the following words and phrases are intended to have the meanings as set out below, except to the extent that the context in which they are used indicates otherwise or are expressly defined to mean something different.
[00041] The symbol “—” means a single bond, “=” means a double bond, “=” means a triple bond. The symbol “^w ” refers to a group on a double bond as occupying each position at the end of the double bond to which the symbol is attached; that is, the geometry, Eor Z-, of the double bond is ambiguous and both isomers are intended to be included. When a group is represented removed from its parent formula, the symbol will be used at the end of the bond which has theoretically been cleaved in order to separate the group from its parent structural formula.
[00042] When chemical structures are represented or described, unless explicitly stated otherwise, all carbons are assumed to have hydrogen substitution to conform to a valence of four. For example, in the structure to the left of the schematic below, there are nine hydrogens involved. The nine hydrogens are represented in the structure on the right. Sometimes a particular atom in the structure is described in the textual formula as having a hydrogen or hydrogens as a replacement (expressly defined hydrogen), eg, -CH2CH2-. It would be understood by a person skilled in the art that the aforementioned descriptive techniques are common chemical techniques to provide brevity and simplicity for the description of otherwise complex structures.

[00043] In this application, some ring structures are represented generically and will be described textually. For example, in the schematic below if ring A is used to describe a phenyl, there are at most four hydrogens in ring A (when R is not H).

[00044] If an R group is represented as "floating" in a ring system, for example in the group:

[00045] thereafter, unless otherwise defined, an R substituent may reside on any atom of the fused bicyclic ring system, excluding the atom bearing the bond with the "^" symbol, as long as a stable structure is formed. In the depicted examples, the R group may reside on an atom in the 5-membered or 6-membered ring of the indolyl ring system.
[00046] When there are more than represented, such as in formulas:

[00047] where there are two groups, namely, the R and the bond that indicates the bond to a precursor structure; then, unless otherwise defined, the "floating" groups may reside on any of the atoms of the ring system, again assuming that each replaces a hydrogen represented, implied, or expressly defined in the ring system and a chemically stable compound would be formed by such an arrangement.
[00048] When an R group is represented as existing in a ring system containing saturated carbons, as for example in the formula:

[00049] where, in this example, y can be more than one, assuming that each replaces a hydrogen currently represented, implied, or expressly defined in the ring; then, unless otherwise defined, two R's can reside on the same carbon. A simple example is when R is a methyl group; there may be a geminal dimethyl on a carbon of the depicted ring (a “ring” carbon). In another example, two R's on the same carbon, which include this same carbon, can form a ring, thus creating a spirocyclic ring structure (a "spirocyclyl" group). Using the previous example, where two R's form, for example, a piperidine ring in a spirocyclic arrangement with cyclohexane, as in the formula:

[00050] “Alkyla” in its broadest sense is intended to include linear, branched, or cyclic hydrocarbon structures, and combinations thereof. Alkyl groups can be fully saturated or with one or more unsaturation units, but not aromatic. In general, alkyl groups are defined by a subscript, a fixed integer or a range of integers. For example, "C8 alkyl" includes n-octyl, iso-octyl, 3-octynyl, cyclohexylethyl, cyclohexylethyl, and others; where the subscript "8" designates that all groups defined by this term have a fixed carbon number of eight. In another example, the term "C1-6 alkyl" refers to alkyl groups having one to six carbon atoms and, depending on any unsaturation, branches and/or rings, the required number of hydrogens. Examples of C1-6 alkyl groups include methyl, ethyl, vinyl, propyl, isopropyl, butyl, s-butyl, t-butyl, isobutyl, isobutenyl, pentyl, pentynyl, hexyl, cyclohexyl, hexenyl, and others. When an alkyl residue having a specific number of carbons is generically mentioned, all geometric isomers having this number of carbons are intended to be covered. For example, "propyl" or "C3 alkyl" each include n-propyl, c-propyl, propenyl, propynyl, and isopropyl. Cycloalkyl is a subset of alkyl and includes cyclic hydrocarbon groups of three to thirteen carbon atoms. Examples of cycloalkyl groups include c-propyl, c-butyl, c-pentyl, norbornyl, norbornenyl, c-hexenyl, adamantyl and others. As mentioned, alkyl refers to alkanyl, alkenyl, and alkynyl residues (and combinations thereof) - it is intended to include, for example, cyclohexyl-methyl, vinyl, allyl, isoprenyl, and others. An alkyl with a particular number of carbons may be mentioned using a more specific but still generic geometric restriction, for example, "C3-6 cycloalkyl" which only means cycloalkyls having between 3 and 6 carbons are intended to be included in this particular definition. Unless otherwise specified, alkyl groups, whether alone or part of another group, for example -C(O)alkyl, are from one to twenty carbons, i.e. C1-20 alkyl. In the example "-C(O)alkyl," where there is no defined carbon count limitation, the carbonyl of the -C(O)alkyl group is not included in the carbon count, as "alkyl" is generically designated. But where a specific carbon limitation is given, for example, in the term "optionally substituted C1-20 alkyl," where the optional substitution includes "oxo" the carbon of any of the carbonyls formed by such "oxo" substitution are included in the carbon counts as these were part of the original carbon count limitation. However, again referring to "optionally substituted C1-20 alkyl," if optional substitution includes groups containing carbon, for example, -CH2CO2H, the two carbons in this group are not included in the carbon limitation of C1-20 alkyl.
[00051] When a carbon number limit is given at the beginning of a term which itself comprises two terms, the carbon number limit is understood to be inclusive of both terms. For example, for the term "C7-14 arylalkyl," both the "aryl" and "alkyl" portions of the term are included in the carbon count, a maximum of 14 in this example, but additional substituent groups thereon are not included in the atom count unless they incorporate a carbon from the group's designated carbon count, as in the “oxo” example above. Likewise when an atom number limit is given, for example "6- to 14-membered heteroarylalkyl," both the "heteroaryl" and the "alkyl" moieties are included in the atom count limitation, but additional substituent groups thereon are not included in the atom count unless they incorporate a carbon from the group's designated carbon count. In another example, "C4-10 cycloalkylalkyl" means a cycloalkyl attached to the parent structure via an alkylene, alkylidene or alkylidine; in this example the group is limited to 10 carbons inclusive of the alkylene, alkylidene or alkylidine subunits. As another example, the "alkyl" portion, for example, of "arylC7-14 alkyl" is intended to include alkylene, alkylidene or alkylidine, unless otherwise stated, for example as in the terms "arylC7-14 alkylene" or "aryla C6-10-CH2CH2-."
[00052] "Alkylene" refers to the bivalent straight, branched and cyclic radical (and combinations thereof) consisting solely of carbon and hydrogen atoms, containing no unsaturation and having from one to ten carbon atoms, eg, methylene , ethylene, propylene, n-butylene and others. Alkylene is like alkyl, referring to the same residues as alkyl, but having two points of attachment and specifically fully saturated. Examples of alkylene include ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), dimethylpropylene (-CH2C(CH3)2CH2-), cyclohexan-1,4-diyl and others.
[00053] "Alkylidene" refers to the bivalent straight, branched, and cyclic (and combinations thereof) unsaturated redicals consisting solely of carbon and hydrogen atoms, having from two to ten carbon atoms, for example, ethylidene, propylidene, n -butylidene, and others. Alkylidene is like alkyl, referring to the same residues as alkyl, but having two points of attachment and specifically at least one double bond unsaturation moiety. Examples of alkylidene include vinylidene (-CH=CH-), cyclohexylvinylidene (-CH=C(C6H13)-), cyclohexen-1,4-diyl and others.
[00054] "Alkydidine" refers to the bivalent straight, branched and cyclic (and combinations thereof) unsaturated radical consisting solely of carbon and hydrogen atoms having two to ten carbon atoms, eg propylid-2-ynyl, n-butylid-1-inyl, and others. Alkylidine is like alkyl, referring to the same residues as alkyl, but having two points of attachment and specifically at least one triple bond unsaturation moiety.
[00055] Any of the above "alkylene," "alkylidene" and "alkylidine" radicals, when optionally substituted, may contain alkyl substitution which itself may contain unsaturation. For example, 2-(2-phenylethynyl-but-3-enyl)-naphthalene (IUPAC name) contains an n-butylid-3-ynyl radical with a vinyl substituent at the 2-position of the radical. Combinations of alkyls and carbon-containing substitutions thereon are limited to thirty carbon atoms.
[00056] "Alkoxy" refers to the group -O-alkyl, where alkyl is as defined herein. Alkoxy includes, by way of example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy, cyclohexyloxy, cyclohexenyloxy, cyclopropylmethyloxy, and others.
[00057] "Haloalkyloxy" refers to the group -O-alkyl, where alkyl is as defined herein, and in addition, alkyl is substituted with one or more halogens. By way of example, a halo-C1-3 alkyloxy group includes -OCF3, -OCF2H, -OCHF2, -OCH2CH2Br, -OCH2CH2CH2I, -OC(CH3)2Br, -OCH2Cl and others.
[00058] "Acyl" refers to the groups -C(O)H, -C(O)alkyl, -C(O)aryl, -C(O)heterocyclyl, -(O)arylalkyl or -C(O) heterocyclylalkyl.
[00059] "α-Amino Acids" refers to the naturally occurring and commercially available α-amino acids and their optical isomers. Typical natural and commercially available α-amino acids are glycine, alanine, serine, homoserine, threonine, valine, norvaline, leucine, isoleucine, norleucine, aspartic acid, glutamic acid, lysine, ornithine, histidine, arginine, cysteine, homocysteine, methionine, phenylalanine, homophenylalanine, phenylglycine, ortho-tyrosine, meta-tyrosine, para-tyrosine, tryptophan, glutamine, asparagine, proline and hydroxyproline. A "side chain of an α-amino acid" refers to the radical found at the α-carbon of an α-amino acid as defined above, for example, hydrogen (for glycine), methyl (for alanine), benzyl (for phenylalanine), etc. .
[00060] “Amino” refers to the group -NH2.
[00061] "Amide" refers to the group -C(O)NH2 or -N(H)acyl.
[00062] "Aryl" (sometimes referred to as "Ar") refers to a monovalent aromatic carbocyclic group, unless otherwise specified, of 6 to 15 carbon atoms having a single ring (eg, phenyl) or multiple condensed rings (eg naphthyl or anthryl) condensed rings which may or may not be aromatic (eg 2-benzoxazolinone, 2H-1,4-benzoxazin-3(4H)-one-7-yl, 9, 9, 10- dihydrophenantrenyl, indanyl, tetralinyl, and fluorenyl and others), provided that the point of attachment is through an atom of an aromatic portion of the aryl group and the aromatic portion at the point of attachment contains only carbons in the aromatic ring. If any portion of the aromatic ring contains a heteroatom, the group is a heteroaryl and not an aryl. Aryl groups are monocyclic, bicyclic, tricyclic or tetracyclic.
[00063] "Arylene" refers to an aryl that has at least two groups attached to it. For a more specific example, "phenylene" refers to a divalent phenyl ring radical. A phenylene, therefore, can have more than two groups attached, but is defined by a minimum of two non-hydrogen groups attached to it.
[00064] "Arylalkyl", sometimes "aralkyl" refers to a residue in which an aryl moiety is attached to a precursor structure via one of an alkylene, alkylidene, or alkylidine radical. Examples include benzyl, phenethyl, phenylvinyl, phenylallyl and others. When specified as "optionally substituted," both the aryl and the corresponding alkylene, alkylidene, or alkylidine portion of an arylalkyl group may be optionally substituted. By way of example, "arylC7-11alkyl" refers to an arylalkyl limited to a total of eleven carbons, for example, a phenylethyl, a phenylvinyl, a phenylpentyl and a naphthylmethyl are all examples of a "arylC7-11alkyl" group ”.
[00065] "Aryloxy" refers to the group -O-aryl, where aryl is as defined herein, which includes, by way of example, phenoxy, naphthoxy, and others.
[00066] "Carboxyl," "carboxy" or "carboxylate" refers to -CO2H or its salts.
[00067] "Carboxylic ester" or "carboxy ester" or "ester" refers to the group -CO2alkyl, -CO2aryl, -CO2heterocyclyl, -CO2arylalkyl or -CO2heterocyclylalkyl.
[00068] "Carbonate" refers to the group -OCO2alkyl, -OCO2aryl, -OCO2heterocyclyl, -OCO2arylalkyl or -OCO2heterocyclylalkyl.
[00069] "Carbamate" refers to the group -OC(O)NH2, -N(H)-carboxyl or -N(H)carboxylic ester.
[00070] “Cyan” or “nitrile” refers to the group -CN.
[00071] "Formyl" refers to the specific acyl group -C(O)H.
[00072] "Halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.
[00073] "Haloalkyl" and "haloaryl" refer generically to alkyl and aryl radicals that are substituted with one or more halogens, respectively. By way of example “dialoaryl,” “dialoalkyl,” “trialoaryl” etc. refer to aryl and alkyl substituted with a plurality of halogens, but not necessarily a plurality of the same halogen; thus 4-chloro-3-fluorophenyl is a dialoaryl group.
[00074] "Heteroalkyl" refers to an alkyl where one or more, but not all, carbons are replaced with a heteroatom. A heteroalkyl group has linear or branched geometry. By way of example, a "2 to 6-membered heteroalkyl" is a group that can contain no more than 5 carbon atoms, because at least one of the maximum 6 atoms must be a heteroatom, and the group is linear or branched. Also, for the purposes of this invention, a heteroalkyl group always starts with a carbon atom, that is, although a heteroalkyl may contain one or more heteroatoms, the point of attachment to the precursor molecule is not a heteroatom. A 2- to 6-membered heteroalkyl group includes, for example, -CH2XCH3, -CH2CH2XCH3, -CH2CH2XCH2CH3, -C(CH2)2XCH2CH3 and others, where X is O, NH, N-C1-6 alkyl and S(O)0-2 , for example.
[00075] "Perhalo" as a modifier means that the group thus modified has all of its available hydrogens replaced with halogens. An example would be "perhaloalkyl." Perhaloalkyls include -CF3, -CF2CF3, perchloroethyl and others.
[00076] "Hydroxy" or "hydroxyl" refer to the -OH group.
[00077] "Heteroatom" refers to O, S, N, or P.
[00078] "Heterocyclyl" in the broadest sense includes aromatic and non-aromatic ring systems and more specifically refers to a stable three- to fifteen-membered ring radical consisting of carbon atoms and one to five heteroatoms. For purposes of this invention, the heterocyclyl radical may be a monocyclic, bicyclic or tricyclic ring system, which may include fused or bridged ring systems as well as spirocyclic systems; and the nitrogen, phosphorus, carbon or sulfur atoms in the heterocyclyl radical can be optionally oxidized to various oxidation states. In a specific example, the group -S(O)0-2-, refers to the -S- (sulfide), -S(O)- (sulfoxide), and -SO2 - (sulfide) bonds. For convenience, nitrogens, particularly but not exclusively those defined as ring aromatic nitrogens, are intended to include their corresponding N-oxide form, although not explicitly defined as such in a particular example. Thus, for a compound having, for example, a pyridyl ring; the corresponding pyridyl-N-oxide is intended to be included in the presently disclosed compounds. Furthermore, ring nitrogen atoms can optionally be quaternized. "Heterocycle" includes heteroaryl and heteroalicyclyl, i.e. a heterocyclic ring may be partially or fully saturated or aromatic. Thus a term such as "heterocyclylalkyl" includes heteroalicyclylalkyls and heteroarylalkyls. Examples of heterocyclyl radicals include, but are not limited to, azetidinyl, acridinyl, benzodioxolyl, benzodioxanyl, benzofuranyl, carbazoyl, cinolinyl, dioxolanyl, indolizinyl, naphthyridinyl, perhydroazepinyl, phenazinyl, phenothiazinyl, phenolazinyl, fenoxa quinoxalinyl, quinolinyl, isoquinolinyl, tetrazoyl, tetrahydroisoquinolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, pyrazolidinyl dihydropyridinyl, tetrahydropyridinyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl, oxazolidinyl, triazolyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, indolylidinyl, isothiazolidinyl, isothiazolidinyl, isothiazolidinyl, isothiazolidinyl, isothiazolidinyl, isothiazolidinyl, isothiazolidinyl, isothiazolidinyl indolyl, octahydroisoindolyl, quino lila, isoquinolyl, decahydroisoquinolyl, benzimidazolyl, thiadiazolyl, benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, diazabicycloheptane, diazapane, diazepine, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyloxy morpholinyloxy, morpholinthia, morpholinthia, sulfolinyloxy, sulfonylthio, sulfolinyloxy
[00079] "Heteroaryl" refers to an aromatic group having 1 to 10 ring carbon atoms and 1 to 4 ring heteroatoms. Heteroaryl groups have at least one aromatic ring component, but heteroaryls can be further unsaturated or partially unsaturated. If any aromatic ring in the group has a heteroatom, then the group is a heteroaryl, even, for example, if other aromatic rings in the group have none of the heteroatoms. For example, 2H-pyrido[3,2-b][1,4]oxazin-3(4H)-one-7-yl, indolyl, and benzimidazolyl are "heteroaryls." Heteroaryl groups can have a single ring (eg pyridinyl, imidazolyl or furyl) or multiple condensed rings (eg indolizinyl, quinolinyl, benzimidazolyl or benzothienyl), where the condensed rings can be aromatic and/or contain a heteroatom or not , provided that the point of attachment to the precursor molecule is through an atom of the aromatic portion of the heteroaryl group. In one embodiment, the nitrogen and/or sulfur ring atom(s) of the heteroaryl group are optionally oxidized to provide the N-oxide (N^O), sulfinyl, or sulfonyl moieties. The compounds described herein contain phosphorus, in a heterocyclic ring or not, including the oxidized forms of phosphorus. Heteroaryl groups are monocyclic, bicyclic, tricyclic or tetracyclic.
[00080] "Heteroaryloxy" refers to -O-heteroaryl.
[00081] "Heteroarylene" generically refers to any heteroaryl that has at least two groups attached to it. For a more specific example, "pyridylene" refers to a divalent pyridyl ring radical. A pyridylene thus can have more than two groups attached, but is defined by a minimum of two non-hydrogen groups attached to it.
[00082] "Heteroalicyclic" specifically refers to a non-aromatic heterocyclyl radical. A heteroalicyclic may contain unsaturation, but it is not aromatic. As mentioned, aryls and heteroaryls are linked to the precursor structure via an aromatic ring. Thus, for example, 2H-1,4-benzoxazin-3(4H)-one-4-yl is a heteroalicyclic, while 2H-1,4-benzoxazin-3(4H)-one-7-yl is an aryl . In another example, 2H-pyrido[3,2-b][1,4]oxazin-3(4H)-one-4-yl is a heteroalicyclic, whereas 2H-pyrido[3,2-b][1 ,4]oxazin-3(4H)-one-7-yl is a heteroaryl.
[00083] "Heterocyclylalkyl" refers to a heterocyclyl group attached to the parent structure via, for example, an alkylene linker. By way of example, the term "heterocyclylC5-14 alkyl" includes groups such as (tetrahydrofuran-3-yl)methyl, (pyridin-4-yl)methyl and 4-(morpholin-4-yl)butan-2-yl, represented below.

[00084] "Heterocyclyloxy" refers to the group -O-heterocyclyl.
[00085] "Heterocyclylalkyloxy" refers to the group -O-heterocyclylalkyl.
[00086] “Nitro” refers to the -NO2 group.
[00087] "Oxo" refers to a double-bonded oxygen radical, =O.
[00088] "Oxy" refers to the radical -(')• (also referred to as —►O), that is, a single-bonded oxygen radical. By way of example, N-oxides are nitrogens that carry an oxy radical.
[00089] When a group with its linking structure is indicated as being linked to two partners; that is, a bivalent radical, eg -OCH2-, then it is understood that each of the two partners may be attached to the particular group at one end, and the other partner is necessarily attached to the other end of the bivalent group, unless explicitly otherwise stated. Stated otherwise, bivalent radicals are not to be interpreted as limited to the orientation depicted, for example "-OCH2-" is intended to mean not only "-OCH2-" as drawn, but also "-CH2O-."
[00090] “Optional” or “optionally” means that the event or circumstance subsequently described may or may not occur, and that the description includes cases where said event or circumstance occurs and cases where it does not. One of ordinary skill in the art would understand that, with respect to any molecule described as containing one or more optional substituents, that only synthetically practicable compounds are intended to be included. "Optionally substituted" refers to all subsequent modifiers in a term, for example in the term "optionally substituted aryl-C1-8 alkyl," optional substitution may occur in either the "C1-8 alkyl" portion or the "aryl" portion ” of the aryl-C1-8 alkyl group. Also, by way of example, optionally substituted alkyl includes optionally substituted cycloalkyl groups. The term "substituted," when used to modify a specified group or radical, means that one or more hydrogen atoms of the specified group or radical are each, independently of one another, substituted with the same or different substituent groups as defined below. . Thus, when a group is defined as "optionally substituted" the definition is intended to cover when the groups are substituted with one or more of the radicals defined below, and when it is not so substituted.
[00091] Substituent groups to substitute in place of one or more hydrogens (any two hydrogens on a single carbon can be substituted with =O, =NR , =N-OR70, =N2 or =S) on standard carbon atoms in the radical group or specified are, unless otherwise specified, -R60, halo, =O, -OR70, -SR70, -N(R80)2, peraloalkyl, -CN, -OCN, -SCN, -NO, - NO2, =N2, -N3, -SO2R70, -SO3-M+, -SO3R70, -OSO2R70, - - + 70 - + - 2+ 70 - + OSO3 M , -OSO3R , -P(O)(O )2( M )2, -P(O)(O )2M , -P(O)(OR )OM , - 70 70 70 70 70 - + 70 P(O)(OR ) 2, -C(O)R , - C(S)R , -C(NR )R , -CO2 M , -CO2R , - C(S)OR70, -C(O)N(R80)2, -C(NR70)(R80)2, -OC (O)R70, -OC(S)R70, -OCO2-M+, 70 70 70 70 70 70 70 - + -OCO2R , -OC(S)OR , -NR C(O)R , -NR C(S) R , -NR CO2 M , - alternatively, two R80's, taken together with the nitrogen atom to which they are attached, form a 3- to 7-membered heteroarylcyclyl which optionally includes 1 to 4 of the same or different additional atoms selected from O , Huh S, of which N is optionally substituted by H or C1-C3 alkyl; and each M+ is a counter ion with a unique net positive charge. Each M+ is independently for each occurrence, for example, an alkaline ion such as K+, Na+, Li+; an ammonium ion, such as +N(R60)4; or an alkaline earth ion, such as [Ca2+]0.5, [Mg2+]0.5, or [Ba2+]0.5 (a “0.5 subscript means, for example, that one of the counter ions for such alkaline earth ions bivalents may be in an ionized form of a compound as described herein and the other typical counter ion such as chloride, or two ionized compounds can serve as the counter ions for such divalent alkaline earth ions, or a doubly ionized compound can serve as the counter ion for such divalent alkaline earth ions). As specific examples, -N(R80)2 is intended to include -NH2, -NH-alkyl, -NH-pyrrolidin-3-yl, N-pyrrolidinyl, N-piperazinyl, 4N-methyl-piperazin-1-yl, N- morpholinyl and others.
[00092] Substituent groups to replace hydrogens on unsaturated carbon atoms in groups containing unsaturated carbons are, unless otherwise specified, -R60, halo, -O-M+, -OR70, -SR70, -S - M+, -N(R80)2, perhaloalkyl, -CN, -OCN, -SCN, -NO, -NO2, -N3, -SO2R70, - - + 70 70 - + 70 -2 + -2 2+ SO3 M , -SO3R , -OSO2R , -OSO3 M , -OSO3R , -PO3 (M )2, -PO3 M , - 70 - + 70 70 70 70 - + P(O)(OR )OM , -P(O) (OR )2, -C(O)R , -C(S)R , -C(NR )R , -CO2 M , - 70 70 80 80 70 80 70 70 CO2R , -C(S)OR , -C (O)NR R , -C(NR )N(R )2, -OC(O)R , -OC(S)R , - + 70 70 70 70 70 70 - -OCO2 M , -OCO2R , -OC (S)OR , -NR C(O)R , -NR C(S)R , -NR CO2 M+ 70 70 70 70 80 70 70 70 , -NR CO2R , -NR C(S)OR , -NR C (O)N(R )2, -NR C(NR )R and - NR70C(NR70)N(R80)2, where R60, R70, R80 and M+ are as previously defined, provided that in the case of substituted alkene or alkyne , the substituents are not -O-M+, -OR70, -SR70, or -S-M+.
[00093] Substituent groups to replace hydrogens on nitrogen atoms in groups containing such nitrogen atoms are, unless otherwise specified, -R60, -O-M+, -OR70, -SR70, -S-M+, -N(R80)2, perhaloalkyl, -CN, -NO, -NO2, -S(O)2R70, -SO3-M+, -SO3R70, -OS(O)2R70, - - + 70 2- + 2-2 + 70 - + OSO3 M , -OSO3R , -PO3 (M )2, -PO3 M , -P(O)(OR )OM , - 70 70 70 70 70 70 70 P(O)(OR )(OR ) , -C(O)R , -C(S)R , -C(NR )R , -CO2R , -C(S)OR , - 80 80 70 80 80 70 70 70 C(O)NR R , -C (NR )NR R , -OC(O)R , -OC(S)R , -OCO2R , - 70 70 70 70 70 70 70 70 OC(S)OR , -NR C(O)R , -NR C (S)R , -NR CO2R , -NR C(S)OR , - 70 80 70 70 70 70 70 80 60 70 NR C(O)N(R )2, -NR C(NR )R and -NR C (NR )N(R )2, where R , R , R80 and M+ are as previously defined.
[00094] In one embodiment, a group that is substituted has 1, 2, 3, or 4 substituents, 1, 2, or 3 substituents, 1 or 2 substituents, or 1 substituent.
[00095] It is understood that in all substituted groups, polymers achieved by defining substituents with other substituents for themselves (for example, substituted aryl having a substituted aryl group as a substituent which is itself substituted with a substituted aryl group, which is further substituted by a substituted aryl group, etc.) are not intended for inclusion here. In such a case that the language allows such multiple substitutions, the maximum number of such substitution interactions is three.
[00096] "Sulfonamide" refers to the group -SO2NH2, -N(H)SO2H, -N(H)SO2alkyl, -N(H)SO2aryl, or -N(H)SO2heterocyclyl.
[00097] "Sulfonyl" refers to the group -SO2H, -SO2alkyl, -SO2aryl, -SO2heterocyclyl, -SO2arylalkyl or -SO2heterocyclylalkyl.
[00098] "Sulfanyl" refers to the group: -SH, -S-alkyl, -S-aryl, -S-heterocyclyl, -S-arylalkyl or -S-heterocyclylalkyl.
[00099] "Sulfinyl" refers to the group: -S(O)H, -S(O)alkyl, -S(O)aryl, -S(O)heterocyclyl, -S(O)-arylalkyl, or -S (O)-heterocyclylalkyl.
[000100] “Suitable starting group” is defined as the term would be understood by a person of ordinary skill in the technique; that is, a group on a carbon, where in the reaction a new bond must be formed, the carbon loses the group in forming the new bond. A typical example using a suitable leaving group is a nucleophilic substitution reaction, for example, on an sp3 hybridized carbon (SN2 or SN1), for example, where the leaving group is a halide, such as a bromide, chloride, fluoride or iodide, the reagent would be the corresponding benzyl halide. Another typical example of such a reaction is the nucleophilic aromatic substitution reaction (SNAr). Another example is an insertion reaction (eg by a transition metal) into partner bonding an aromatic reaction which carries a leaving group followed by the reductive bond. “Suitable starting group” is not limited to such mechanical restrictions. Examples of suitable leaving groups include halogens, optionally substituted aryl or alkyl sulfonates, phosphonates, azides and -S(O)0-2R where R is, for example optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl. Those skilled in the art of organic synthesis will easily identify suitable leaving groups to carry out a desired reaction under different reaction.
[000101] "Stereoisomers" and "stereoisomers" refer to compounds that have the same atomic connectivity but different atomic arrangement in space. Stereoisomers include cis-trans isomers, E and Z isomers, enantiomers and diastereomers. Compounds as described herein, or pharmaceutically acceptable salts thereof, may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms which may be defined, in terms of absolute stereochemistry, as (R)- or ( S)- or, as (D)- or (L)- for amino acids. The present invention is intended to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers can be prepared using chiral synthons, chiral reagents, or resolved using conventional techniques, such as by : formation of diastereoisomeric salts or complexes which can be separated, for example, by crystallization; by forming diastereoisomeric derivatives which can be separated, for example, by crystallization, selective reaction of an enantiomer with an enantiomer-specific reagent, for example oxidation or enzymatic reduction, followed by separation of the modified and unmodified enantiomers; or gas-liquid chromatography or liquid chromatography in a chiral environment, for example on a chiral support such as silica with an attached chiral ligand or in the presence of a chiral solvent. It will be appreciated that where a desired enantiomer is converted to another chemical entity by one of the separation procedures described above, another step may be required to release the desired enantiomeric form. Alternatively, specific enantiomers can be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by conversion from one enantiomer to the other by asymmetric transformation. For a mixture of enantiomers, enriched in a particular enantiomer, the larger component enantiomer can be further enriched (with a concomitant loss in yield) by recrystallization.
[000102] When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise specified, it is intended that the compounds include both E and Z geometric isomers.
[000103] "Tautomer" refers to alternate forms of a molecule that differ only in the electronic bonding of atoms and/or in the position of a proton, such as enol-keto and imine-enamine tautomers, or the tautomeric forms of heteroaryl groups containing a -N=C(H)-NH- ring atom arrangement, such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles. A person of ordinary skill in the art would recognize that other tautomeric ring atom arrangements are possible and considered here.
[000104] "Meta" for the purposes of this invention refers to the position of a substituent on a phenyl or a six-membered heteroaryl ring relative to another substituent on the ring; the relative position being the substitution 1,3. That is, starting with a substituent as being attached to a first atom of the six-membered ring and, counting the atoms inclusive of the first atom, another substituent is on atom 3 of the six-membered ring, the relative orientation of the substituents around the six-membered ring is "meta." For example, compound J, depicted below, has a methyl group "meta" relative to N2 of pyrimidinediamine; compounds K1 and K2 also have a "meta" methyl group relative to N2 of pyrimidinediamine. Thus, in some cases, instead of using specific ring atom numbering, the term “meta” is used. In such cases both meta isomers are intended to be included.

[000105] "For" for the purposes of this invention refers to the position of a substituent on a phenyl or a six-membered heteroaryl ring relative to another substituent on the ring; the relative position being 1,4- substitution. That is, starting from a substituent as being attached to a first atom of the six-membered ring and, counting atoms inclusive of the first atom, another substituent is on atom 4 of the six-membered ring, the relative orientation of the substituent around the ring. of six members is "for." For example, compound L, depicted below, has a methyl group "para" with respect to N2 of pyrimidinediamine; compound M also has a methyl group "para" with respect to N2 of pyrimidinediamine. Thus, in some cases, instead of using specific ring atom numbering, the term “for” is used.

[000106] "Patient" or "Individual" refers to mammals and other animals, particularly humans. Thus the methods are applicable in both human therapy and veterinary applications. In one embodiment the patient or subject is a mammal. In another embodiment the patient or individual is a human being.
[000107] "Pharmaceutically acceptable salt" refers to the pharmaceutically acceptable salts of a compound, which salts are derived from a variety against organic and inorganic ions well known in the art and include, by way of example only, sodium, potassium, calcium , magnesium, ammonium, tetraalkylammonium, and others; and when the molecule contains a basic functionality, the salts of organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate, and others. Pharmaceutically acceptable acid addition salts are those salts which retain the biological effectiveness of the free bases whilst being formed by acidic partners which are not biologically or otherwise undesirable, for example the inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid , nitric acid, phosphoric acid, and others, as well as organic acids such as acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, acid citric, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, salicylic acid, xinafoic acid (1-hydroxy-2-naphthoic acid), pamoic acid (also called embonic acid or 4,4'-methylonobis(3-hydroxy-2-naphthoic acid) and others. Pharmaceutically acceptable base addition salts include those derived from inorganic bases such as salts of sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum and others. Exemplary salts are the ammonium, potassium, sodium, calcium, and magnesium salts. Salts of the presently disclosed compounds can be derived from pharmaceutically acceptable non-toxic organic bases include, but are not limited to salts of primary, secondary, and tertiary amines, substituted amines which include naturally occurring substituted amines, cyclic amines and ion exchange resins basic such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, 2-amino-2-hydroxymethyl-propane-1,3-diol ("Tris" salt), dicyclohexylamine, lysine , arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and others. (See, for example, S.M. Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977; 66: 1-19 which is incorporated herein by reference).
[000108] "Pharmaceutically acceptable amount" and "therapeutically effective amount" refer to an amount of a compound sufficient to treat a specified disorder or disease or one or more of its symptoms and/or to prevent the occurrence of the disease or disorder. The amount of a compound that constitutes a "therapeutically effective amount" will vary depending on the compound, the disease state and its severity, the age of the patient being treated, and so on. The therapeutically effective amount can be routinely determined by a person of ordinary skill in the art.
[000109] "Prodrug" refers to compounds that are transformed in vivo to produce the parent compound, for example, by hydrolysis in the intestine or enzymatic conversion in the blood. Common examples include, but are not limited to ester and amide forms of a compound having an active form that carries a carboxylic acid moiety. Examples of pharmaceutically acceptable esters of the compounds of this invention include, but are not limited to, alkyl esters (for example having between about one and about six carbons) where the alkyl group is a straight or branched chain. Acceptable esters also include cycloalkyl esters and arylalkyl esters such as, but not limited to, benzyl. Examples of pharmaceutically acceptable amides of the compounds of this invention include, but are not limited to, primary amides, and secondary and tertiary alkyl amides (for example having between about one and about six carbons). Amides and esters of the compounds of the present invention can be prepared according to conventional methods. A full debate on prodrugs is provided in T. Higuchi and V. Stella, “Pro-drugs as Novel Delivery Systems,” Vol 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference for all purposes.
[000110] “Metabolite” refers to the decomposition or end product of a compound or its salt produced by metabolism or biotransformation in the animal or human body; for example, biotransformation to a more polar molecule such as by oxidation, reduction, or hydrolysis, or to a conjugate (see Goodman and Gilman, "The Pharmacological Basis of Therapeutics" 8th Ed., Pergamon Press, Gilman et al. (eds. ), 1990 which is incorporated herein by reference). The metabolite of a compound described herein or its salt may itself be a biologically active compound in the body. Although a prodrug described herein would meet these criteria, i.e., would form a described in vivo biologically active precursor compound, "metabolite" is intended to encompass these compounds not considered to have lost a progroup, but rather all other compounds that are formed in vivo upon administration of a compound as described herein that retains the biological activities described herein. Thus one aspect of the specifically disclosed 2,4-pyrimidine diamine compounds considered herein is a metabolite of a compound described herein. For example, a biologically active metabolite is serendipitously discovered, that is, no pro-drug planning per se has been undertaken. Otherwise stated, biologically active compounds inherently formed as a result of practicing the methods as described herein are considered and disclosed herein. "Solvate" refers to a complex formed by combining solvent molecules with solute molecules or ions. The solvent can be an organic compound, an inorganic compound, or a mixture of both. Some examples of solvents include, but are not limited to, methanol, N,N-dimethylformamide, tetrahydrofuran, dimethyl sulfoxide, and water. The compounds described herein can exist in unsolvated form as well as solvated with solvents, pharmaceutically acceptable or not, such as water, ethanol, and others. Solvated forms of the presently disclosed compounds are contemplated herein and are encompassed by the invention, at least in general terms.
[000111] "Treating" or "treatment" as used herein encompasses the treatment of the disease or condition of interest in a mammal, preferably a human, having the disease or condition of interest, and includes: (i) preventing the disease or condition to occur in a mammal, in particular, when such a mammal is predisposed to the condition but has not yet been diagnosed as having the same; (ii) inhibit the disease or condition, for example, arrest or slow down its development; (iii) alleviating the disease or condition, for example, causing the regression of the disease or condition or a symptom thereof; or (iv) stabilize the disease or condition.
[000112] As used herein, the terms "disease" and "condition" may be used interchangeably or may be different in that the particular illness or condition may not have a known causative agent (so the etiology has not yet been developed) and therefore it is not yet recognized as a disease but only as an unwanted condition or syndrome, where a more or less specific set of symptoms has been identified by doctors.
[000113] Similarly, it is understood that the above definitions are not intended to include non-permissible substitution patterns (eg methyl substituted with 5 fluorine groups). Such impermissible substitution patterns are easily recognized by a person having ordinary skill in the technique. Compounds and Compositions
[000114] Disclosed herein are novel 2,4-pyrimidinediamine compounds, prodrugs of the compounds, methods of making the compounds, and methods of using these compounds in treating conditions where JAK pathway targeting or modulation, which includes inhibition, of JAK kinases, particularly JAK3, are therapeutically useful. These conditions include, but are not limited to, leukemia, lymphoma, transplant rejection (e.g. pancreatic islet transplant rejection, heart transplant rejection, kidney transplant rejection, liver transplant rejection, heart transplant rejection, lung), bone marrow transplant applications (eg, graft-versus-host disease), autoimmune diseases (eg, diabetes), and inflammation (eg, asthma, allergic reactions, eye disorders). Given the severity and prevalence of these diseases and conditions, new therapies are needed. Compounds
[000115] The compounds, and their salts, described herein are generally pyrimidine 2,4-diamines, substituted at the 5-position with various groups; substituted on the 2-amine with various optionally substituted aromatic groups; and substituted on the 4-amine with one of a benzo[d]oxazol-2(3H)-one, a 1H-benzo[d]imidazol-2(3H)-one, a benzo[d]thiazol-2(3H) - one, a benzo[d][1,3]dithiol-2-one, a benzo-[d][1,3]oxathiol-2-one, a benzo[d][1,3]dioxol-2- one, a [1,3]-oxathiolo-[4,5-b]pyridin-2-one, a thiazolo[5,4-b]pyridin-2(1H)-one, an oxazolo[5,4-b ]pyridin-2(1H)-one, a [1,3]oxathiolo[5,4-b]pyridin-2-one, a thiazolo[4,5-b]pyridin-2(3H)-one, an oxazole [4,5-b]pyridin-2(3H)-one, one [1,3]dioxolo[4,5-b]pyridin-2-one, one [1,3]dithiolo[4,5-b] -pyridin-2-one, a 1H-imidazo[4,5-b]pyridin-2(3H)-one, a [1,3]-oxathiolo[4,5-b]pyrazin-2-one, a thiazole [5,4-b]pyrazin-2(3H)-one, one oxazolo[5,4-b]pyrazin-2(3H)-one, one [1,3]dioxolo[4,5-b]pyrazin- 2-one, a [1,3]dithiolo[4,5-b]pyrazin-2-one and a 1H-imidazo[4,5-b]pyrazin-2(3H)-one; each optionally substituted with one or more groups that include prodrug moieties as described herein. In addition to the groups described above, the N2- and N4-amines of the pyrimidinediamine system may also have optionally substituted alkyl groups and/or prodrug groups.
[000116] More specifically, the compounds are described in terms of formula I:
Where:
[000117] X and Y are each independently O, S, S(O), SO2 or NR1;
[000118] each R1 is independently for each occurrence H, optionally substituted C1-6 alkyl, C(O)-C1-6 alkyl, CO2-C1-6 alkyl or R50;
[000119] each R50 is -C(R9)2-A-R10, where A is O or S; each R9 is independently for each occurrence H, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl, or optionally substituted C7-16 arylalkyl; or alternatively, two R 9 , together with the carbon to which they are attached, form an optionally substituted C 3-8 cycloalkyl group or an optionally substituted 3- to 8-membered heteroarylcyclyl group; R10 is Ra, -P(O)(OR11)2 -P(O)(OR11)N(R12)2 or -P(O)(N(R12)2)2 ; each R11 is independently for each occurrence Ra or a monovalent cationic group; either two R11 together with the atoms to which they are attached form a 4- to 8-membered cyclic phosphate group, or two R11 together represent a divalent cationic group; each R12 is independently for each occurrence Rc or -C1-3-alkylN(Rc)2; or two R12, each on nitrogens separate from -P(O)(N(R12)2)2, together with the atoms to which they are attached, form a 4- to 8-membered cyclic phosphonic acid bisamide group; or an R12 together with R11, of the group -P(O)(OR11)N(R12)2, together with the atoms to which they are attached, form a 4- to 8-membered cyclic phosphonamidate group;
[000120] ring A is a C6-10 aryl or a 5- to 10-membered heteroaryl;
[000121] each R2 is independently for each occurrence H, Re, Rb, Re substituted with one or more of the same or different Ra and/or Rb, -ORe substituted with one or more of the same or different Ra and/or Rb, - SRe substituted with one or more of the same or different Ra and/or Rb, -C(O)Re substituted with one or more of the same or different Ra and/or Rb, -N(Ra)Re where Re is substituted with one or more of the same or different Ra and/or Rb, - S(O)2Re replaced with one or more of the same or different Ra and/or Rb, - N(Ra)-S(O)2Re where Re is replaced with one or more of the same or different Ra and/or Rb, -B(ORa)2, -B(N(Rc)2)2, -(C(Ra)2)m-Rb, -O-(C(Ra)2 )m-Rb, -abba ab S-(C(R)2)mR, -O-(C(R)2)mR, -N(R)-(C(R)2)mR, -O -(CH2)m-bb aabaa CH((CH2)mR)R, -C(O)N(R)- (C(R)2)mR, -O-(C(R)2)mC(O) N(R )- ab ab aaab (C(R )2)mR , -N((C(R )2)mR )2, -S-(C(R )2)mC(O)N(R )- (C(R )2)mR , - aaabaa ab N(R )-C(O)-N(R )-(C(R )2)mR , -N(R )-C(O)-(C( R )2)mC(R)(R)2 or -N(Ra)-(C(Ra)2)mC(O)-N(Ra)- (C(Ra)2)m-Rb;
[000122] each Ra is independently for each occurrence H, deuterium, C1-6 alkyl, C3-8 cycloalkyl, C4-11 cycloalkylalkyl, C6-10 aryl, C7-16 arylalkyl, 2 to 6-membered heteroalkyl, 3 to 3-membered heteroalicyclyl 10-membered, 4- to 11-membered heteroalicyclylalkyl, 5- to 15-membered heteroaryl, or 6- to 16-membered heteroarylalkyl;
[000123] each Rb is independently for each occurrence =O, -ORa, -O-(C(Ra)2)m-ORa, halo-C1-3 alkyloxy, =S, -SRa, =NRa, =NORa, - N(Rc)2, halo, -CF3, -CN, -NC, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)Ra, -S(O)2Ra, - SO3Ra, -S(O)N(Rc)2, -S(O)2N(Rc)2, -OS(O)Ra, -OS(O)2Ra, -OSO3Ra, - OS(O)2N(Rc) 2, -C(O)Ra, -CO2Ra, -C(O)N(Rc)2, -C(NRa)-N(Rc)2, -C(NOH)-Ra, -C(NOH)-N (Rc)2, -OC(O)Ra, -OC(O)ORa, -OC(O)N(Rc)2, -OC(NH)-N(Rc)2, -ac aaaaa OC(NR )- N(R )2, -N(R )-S(O)2H, -[N(R )C(O)]nR , -[N(R )C(O)]nOR , - [N(Ra) C(O)]nN(Rc)2 or -[N(Ra)C(NRa)]nN(Rc)2;
[000124] each Rc is independently for each occurrence Ra, or, alternatively, two Rc are taken together with the nitrogen atom to which they are attached to form a 3- to 10-membered heteroallylcyclyl or a 5- to 10-membered heteroaryl that can optionally including one or more of the same or different additional heteroatoms and which is optionally substituted with one or more of the same or different Ra and/or Rd groups;
[000125] each Rd is =O, -ORa, halo-C1-3 alkyloxy, C1-6 alkyl, =S, -SRa, =NRa, =NORa, -N(Ra)2, halo, -CF3, -CN , -NC, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)Ra, -S(O2)Ra, -SO3Ra, -S(O)N(Ra)2, -S(O)2N(Ra)2, -OS(O)Ra, - OS(O)2Ra, -OSO3Ra, -OS(O)2N(Ra)2, -C(O)Ra, -CO2Ra, - C(O)N(Ra)2, -C(NRa)N(Ra)2, -C(NOH)Ra, -C(NOH)N(Ra)2, -OCO2Ra, -OC(O)N(Ra )2, - aa aaaaaa OC(NR )N(R )2, -[N(R )C(O)]nR , -(C(R )2)n-OR , -N(R )-S(O )2R , -C(O)-C1-6 haloalkyl, -S(O)2C1-6 haloalkyl, -OC(O)Ra, -O(C(Ra)2)m -ORa, -a aa a aa a S(C(R )2)m-OR , -N(R )C 1-6 haloalkyl, -P(O)(OR )2, -N(R )-(C(R )2)m -OR , - [N(Ra)C(O)]nORa, -[N(Ra)C(O)]n N(Ra)2, -[N(Ra)C(NRa)]nN(Ra)2 or - N( Ra)C(O)haloC1-6alkyl; two Rd, taken together with the atom or atoms to which they are attached, combine to form a partially or fully saturated 3- to 10-membered mono or bicyclic ring, optionally containing one or more heteroatoms and optionally substituted with one or more Ra ;
[000126] each Re is independently for each occurrence C1-6 alkyl, C3-8 cycloalkyl, C4-11 cycloalkylalkyl, C6-10 aryl, C7-16 arylalkyl, 2 to 6 membered heteroalkyl, 3 to 10 membered heteroalicyclyl, heteroalicyclylalkyl 4- to 11-membered, 5- to 15-membered heteroaryl, or 6- to 16-membered heteroarylalkyl;
[000127] p is 0, 1, 2, 3 or 4;
[000128] each m is 1, 2 or 3;
[000129] each n is 0, 1, 2 or 3;
[000130] two R2 groups, taken together with the atom or atoms to which they are attached, combine to form a partially or fully saturated 4- to 10-membered mono or bicyclic ring, optionally containing one or more heteroatoms and optionally substituted with one or more Ra and/or Rb;
[000131] Z1 and Z2 are each independently CH, CR2 or N;
[000132] R3 is H, optionally substituted C1-6 alkyl or R50;
[000133] R4 is H, optionally substituted C1-6 alkyl or R50; and
[000134] R5 is H, halo, -CN, optionally substituted C1-6 alkyl, alkynyl, hydroxy, optionally substituted C1-6 alkoxy, nitro, -N(Ra)2, -C(O)N(Ra)2, -CO2Ra or -C(O)Ra.
[000135] In one embodiment, structural formula I includes compounds where ring A is a phenyl or a pyridyl substituted with one or more groups. In one embodiment, ring A is a phenyl with at least one para group relative to N2 of pyrimidinediamine. In another embodiment, ring A is a pyridyl with at least one para group relative to the N2 of the pyrimidinediamine. In a more specific embodiment, ring A is a pyridin-3-yl (where N2 of pyrimidinediamine is in the 3-yl position) with at least one para group relative to N2 of pyrimidinediamine. In another more specific embodiment, ring A is a pyridin-2-yl (where N2 of the pyrimidinediamine is in the 2-yl position) with at least one para group relative to the N2 of the pyrimidinediamine. In other embodiments, one or two meta groups may replace or augment the para group in the embodiments described above. In all of the above embodiments, groups in the para and/or I position may include nitrogen, for example an optionally substituted amine, directly attached to ring A or in the same embodiments attached to ring A via an alkylene. Such optionally substituted amines include those defined by -N(Rc)2 as in relation to formula I. In a specific embodiment, an optionally substituted amine is attached to ring A via a C1-6 alkylene. In a still more specific embodiment, the optionally substituted amine is attached to ring A via a C 1-6 alkylene and the amine, -N(Rc) 2 as in relation to formula I, is itself substituted with a -N(Rc)2 group.
[000136] As mentioned, certain compounds disclosed herein have structural formula I where ring A is a phenyl substituted with one or more R2 groups. Thus, in one disclosed compounds have the formula IA:

[000137] where the variables are as described in formula I, and further: R1 is H, optionally substituted C1-6 alkyl or R50; each of R2a, R2b, R2c, R2d and R2e is independently for each occurrence as defined for R2; and R5 is H, halo, -CN, optionally substituted C1-6 alkyl, nitro, -N(Ra)2, -C(O)N(Ra)2, -CO2Ra or -C(O)Ra.
[000138] In one embodiment, R2e is halo or optionally substituted C1-6 alkyl. In another embodiment, R2e is halo or methyl. In another embodiment, R2e is F or methyl.
[000139] In one embodiment, R2e and R4 are both H. One such embodiment is a compound of the formula IB:

[000140] where R1 is H or R50; each of R2a, R2b, R2c and R2d is independently for each occurrence as defined for R2 in formula I; and R5 is H, halo, -CN, C1-6 alkyl, nitro, -N(Ra)2, -C(O)N(Ra)2, -CO2Ra or -C(O)Ra. In one embodiment, R5 is H, halo, -CN, C1-6 alkyl, nitro, -C(O)N(Ra)2, -CO2Ra or -C(O)Ra. In another embodiment, R5 is H, halo or C1-6 alkyl. In another embodiment, R5 is H, halo or methyl. In another embodiment, R5 is halo or methyl. In another embodiment, R5 is F or CH3. In another embodiment, R5 is CH3.
[000141] In one embodiment of the compounds of formula IB as described above, each of R2a, R2b and R2c is independently H, C1-6 alkyl, -OC1-6 alkyl, -OCF3, -N(H) C1 alkyl -6, -N(C1-6 alkyl)2, halo, -OCF2H, -OCH2F, -CF3, -CN, -C(O)Ra, -CO2Ra, -C(O)N(Rc)2, -O (C(Ra)2)m-Rb or -(C(Ra)2)m-Rb; and R2d is H or F; where at least one of R2a, R2b and R2c is not H. In another embodiment, each of R2a, R2b and R2c is independently H, C1-4 alkyl, -OC1-4 alkyl, -OCF3, -N( H) C1-4 alkyl, -N(C1-4 alkyl)2, halo, -OCF2H, -OCH2F, -CF3, -CN, -C(O)H, -C(O) C1-4 alkyl, -CO2Ra , -C(O)N(Rc)2, -O(CH2)2-ORa or -(CH2)1-2-Rb; and R2d is H or F; where at least one of R2a, R2b and R2c is not H. In another embodiment, each of R2a, R2b and R2c is independently H, C1-2 alkyl, -OC1-2 alkyl, -OCF3, -N( H) C1-2 alkyl, -N(C1-2 alkyl)2, halo, -OCF2H, -OCH2F, -CF3, -CN, -CO2Ra, -C(O)N(Rc)2, -O(CH2) 2-O-C 1-2 alkyl or -(CH2)1- 2-OH; and R2d is H or F; where at least one of R2a, R2b and R2c is not H. In yet another embodiment, each of R2a, R2b and R2c is independently C1-2 alkyl, -OC1-2 alkyl, -OCF3, -N(H ) C1-2 alkyl, - N(C1-2 alkyl)2, halo, -OCF2H, -OCH2F, -CF3, -CN, -CO2Ra, -C(O)N(Rc)2, - O(CH2)2 -OC 1-2 alkyl or -(CH2)1-2-OH; and R2d is H or F; where at least one of R2a, R2b and R2c is not H. In another embodiment, each of R2a, R2b and R2c is independently CH3, -OCH3, -OCF3, -N(H) C1-2 alkyl, - N(C 1-2 alkyl), halo or -CF3; and R2d is H or F; where at least one of R2a, R2b and R2c is not H. In another embodiment, each of R2a, R2b and R2c is independently CH3, -OCH3, -OCF3, -N(H) C1-2 alkyl, - N(C1-2alkyl)2, halo or -CF3; and R2d is H or F; where at least one of R2a, R2b and R2c is not H.
[000142] For each of the embodiments described above for the compounds of formula IB, there are embodiments where: 1) R2a is H, and R2b and R2c are each independently one of the groups other than H described, 2) R2b is H, and R2a and R2c are each independently one of the described non-H groups, 3) R2c is H, and R2a and R2b are each independently one of the described non-H groups, 4) R2a and R2b are H, and R2c is one of the non-H groups described, 5) 4) R2a and R2c are H, and R2b is one of the non-H groups described, 6) 4) R2b and R2c are H, and R2a is one of the non-H groups described, 7) R2a , R2b and R2c are each independently one of the non-H groups described.
[000143] According to embodiment 1) above, one embodiment is a compound where R2a is H; R2b is C1-3 alkyl, F or -OC13 alkyl; R2c is -OC1-3 alkyl; and R2d is H or F. According to embodiment 1) above, one embodiment is a compound where R2a is H; R2b is C1-3 alkyl; R2c is -OC1-3 alkyl; and R2d is H or F. According to embodiment 1) above, one embodiment is a compound where R2a is H; R2b is C1-3 alkyl; R2c is -OC1-3 alkyl; and R2d is F. According to embodiment 1) above, one embodiment is a compound where R2a is H; R2b is CH3; R2c is -OCH3; and R2d is F. According to embodiment 1) above, one embodiment is a compound where R2a is H; R2b is -OC1-3 alkyl; R2c is -OC1-3 alkyl; and R2d is H or F. According to embodiment 1) above, one embodiment is a compound where R2a is H; R2b is -OC1-3 alkyl; R2c is -OC1-3 alkyl; and R2d is F. According to embodiment 1) above, one embodiment is a compound where R2a is H; R2b is -OCH3; R2c is -OCH3; and R2d is F.
[000144] According to embodiment 7) above, one embodiment is a compound where R2a, R2b and R2c are each independently CH3, -OCH3, or F; and R2d is H or F. According to embodiment 7) above, one embodiment is a compound where two of R2a, R2b and R2c are CH3; the other of R2a, R2b and R2c is F; and R2d is H or F. According to embodiment 7) above, one embodiment is a compound where two of R2a, R2b and R2c are CH3; the other of R2a, R2b and R2c is -OCH3; and R2d is H or F. According to embodiment 7) above, one embodiment is a compound where two of R2a, R2b and R2c are -OCH3; the other of R2a, R2b and R2c is F; and R2d is H or F. According to embodiment 7) above, one embodiment is a compound where two of R2a, R2b and R2c are -OCH3; the other of R2a, R2b and R2c is CH3; and R2d is H or F. According to embodiment 7) above, one embodiment is a compound where one of R2a, R2b and R2c is CH3; one of R2a, R2b and R2c is -OCH3; and one of R2a, R2b and R2c is F; and R2d is H or F.
[000145] Another embodiment is a compound according to formula IB, where two of R2a, R2b and R2c are each independently H, C1-6 alkyl, -OC1-6 alkyl, -OCF3, halo, -OCF2H , -OCH2F, -CF3, -CN, -O(C(Ra)2)m-Rb or -(C(Ra)2)m-Rb; one of R2a, R2b and R2c is:


[000146] optionally substituted with one or more of the same or different Ra and/or Rb groups; and R2d is H or F. Another embodiment is a compound according to formula IB, where two of R2a, R2b and R2c are each independently H, C1-6 alkyl, -OC1-6 alkyl, -OCF3, halo, -OCF2H, -OCH2F, -CF3, -CN, -O(C(Ra)2)m-Rb or -(C(Ra)2)m-Rb; one of R2a, R2b and R2c is:

[000147] optionally substituted with one or more of the same or different groups Ra and/or Rb, and R2d is H or F.
[000148] Another embodiment is a compound according to formula IB, where two of R2a, R2b and R2c are each independently H, C1-6alkyl, -OC1-6alkyl, -OCF3, halo, -OCF2H, - OCH2F, -CF3, -CN, -O(C(Ra)2)m-Rb or -(C(Ra)2)m-Rb; one of R2a, R2b and R2c is a water-solubilizing group; and R2d is H or F. A water-solubilizing group is a group which is of sufficient hydrophilic character to improve or increase the water solubility of the compound in which it is included, as compared to an analogous compound which does not include what it does not. includes the group. The hydrophilic character can be achieved, for example, by the inclusion of functional groups that ionize under the conditions of use to form charged moieties (for example, carboxylic acids, sulfonic acids and salts, and phosphoric acids and salts, amines, etc.) ; groups that include permanent charges (eg, quaternary ammonium groups); and/or heteroatoms or heteroatomic groups. For example, -O-(C(Ra)2)m-Rb, -S-(C(Ra)2)m-Rb, -Ob aaab bb -(C(R)2)mR , -N(R ) -(C(R)2)mR, -O-(CH2)m-CH((CH2)mR)R, -C(O)N(Ra)-(C(Ra)2)m-Rb and -N ((C(Ra)2)mRb)2. More specific examples include -O-C1-6 alkylene-Rb, -S-C1-6 alkylene-Rb, -O-C1-6 alkylene-Ra where Ra is heterocyclyl, -N(Ra)-C1-6 alkylene- Rb, -O-C1-6 alkylene-CH((CH2)1-2Rb)Rb, -C(O)N(Ra)-C1-6 alkylene-Rb and -N((C(Ra)2)1- 3Rb)2. Even more specific examples include -O-C1-4 alkylene-Rb, -S-C1-4 alkylene-Rb, -O-C1-4 alkylene-Ra where Ra is heterocyclyl, -N(H)-C1-4 alkylene -Rb, -O- C1-4 alkylene-CH((CH2)1-2Rb)Rb, -C(O)N(H)-C1-4 alkylene-Rb and -N((CH2)1-3Rb)2 . In another specific example, according to the formula given above for water-solubilizing groups, the water-solubilizing group is an amino acid attached to the molecule via a bond to the amino acid's nitrogen. In a more specific example, a water solubilization group is an α-amino acid or derivative thereof linked to the precursor ring, for example, the A ring and/or in Z1 or Z2, through the nitrogen of the α-amino acid, for example - N(H)C(Ra)2-Rb, where Rb is -CO2Ra or -C(O)N(Rc)2. In another specific embodiment, the water-solubilizing group is morpholino, piperidinyl, N-C 1-6 alkyl piperidinyl, piperazinyl, N-C 1-6 alkyl piperazinyl, pyrrolidinyl, N-C 1-6 alkyl pyrrolidinyl, diazepinyl, N -C 1-6 alkyl azepinyl, homo-piperazinyl, N-C 1-6 alkyl homopiperazinyl, imidazoyl, and others. In another example the water-solubilizing group is one of the aforementioned rings attached to the precursor molecule via an alkylene, alkylidene, alkylidine linker. In a more specific embodiment, the water-solubilizing group is one of the aforementioned rings attached to the parent molecule via a C1-6 alkylene, where one or two of the alkylene carbons is independently replaced with one of O , S or NH, but not where any two of the aforementioned heteroatoms are contiguous in the linker. Other water-solubilizing groups are well known and include, by way of example, hydrophilic groups such as alkyl or heteroalicyclyl groups substituted with one or more of an amine, alcohol, a carboxylic acid, a phosphorous acid, a sulfoxide, a carbohydrate , a sugar alcohol, an amino acid, a thiol, a polyol, an ether, a thioether, and a quaternary amine salt.
[000149] Another embodiment is a compound according to formula I, of structural formula IC:

[000150] where R1 is H or R50; R5 is H, halo, -CN, C1-6 alkyl, nitro, -N(Ra)2, -C(O)N(Ra)2, -CO2Ra or -C(O)Ra; R20 is H or C1-6 alkyl; each R15 is independently H or C1-6 alkyl, or two of R15 together on the same carbon are oxo; and G is O or NH. In another embodiment, R5 is H, halo, -CN, C1-6 alkyl or nitro. In another embodiment, R5 is H, halo or C1-6 alkyl. In another embodiment, R5 is H, halo or methyl. In another embodiment, R5 is halo or methyl. In another embodiment, R5 is F or CH3. In another embodiment, R5 is CH3.
[000151] For each of the above embodiments of the compounds of structural formula I, IA, IB and IC, there is an embodiment where R1 is H or R50; R50 is -CH2OP(O)(OR11)2; and each R11 is independently for each occurrence Ra or a monovalent cationic group; either two R11 together with the atoms to which they are attached form a 4- to 8-membered cyclic phosphate group, or two R11 together represent a divalent cationic group. Also, for each of these embodiments, this is a more specific embodiment where each R11 is independently for each occurrence H, t-butyl, or a pharmaceutically acceptable cation such as HOCH2CH2N(CH3)3+, Na+, Li+ or K+.
[000152] As mentioned, the 2,4-pyrimidinediamine compounds and prodrugs, as well as their salts, can also be in the form of hydrates, solvates, and N-oxides, as is well known in the art. One embodiment is a pharmaceutically acceptable salt form of a compound of formula I. The pharmaceutically acceptable salts of the present invention may be formed by conventional means, such as by reacting the free base form of the product with one or more equivalents of the product. appropriate acid in a solvent or medium in which the salt is insoluble or in a solvent such as water that is removed in a vacuum, by freeze drying, or by exchanging the anions of an existing salt for another anion in an ion exchange resin proper. The present invention includes within its scope solvates of its compounds and salts and hydrates of 2,4-pyrimidinediamine, for example, a hydrated formate salt.
[000153] One embodiment is a compound selected from Tables I and II, or a pharmaceutically acceptable stereoisomer, tautomer, prodrug, solvate, or salt thereof. Many compounds described herein were manufactured as the precursor, at least one salt form, or both the precursor and one or more salts. Some specific manufactured salts, alluded to by their Table I and II designations, include the arginine salt: compound I-130; the calcium salt: compound I-118; the Tris salt: compound I132; the dipotassium salt: compound I-136; the formate salt: compounds I-1, I8, I-16, I-67, I-84 through I-87, I-157 through I-166, I-196, I-197, I-207 through I- 209, II-4, II-5 and II-23 through II-25; the diformate salt: compounds II-2 and II-3; the monotrifluoroacetate salt: compounds I-2 through I-7, I-10, I-11; I-13 through I-15, I-18, I20 through I-22, I-24, I-29, I-32, I-35 through I-45, I-47 through I-49, I-51, I-54 to I-61, I64 to I-66, I-68, I-69, I-71 to I-83, I-109 to I-112, I-116, I-117, I-120 to I127, I-129, I-167, I-168, I-171 to I-185, I-187 to I-195, I-198 to I-202, II-1, II-6, II-7 and II-26 to II-28; the ditrifluoroacetate salt: compounds I-89, I-90, I95, I-97, I-103, I-104, I-169, I-170, I-186 and II-29 through II-31; the benzene sulphonic acid salt: compounds I-107, I-108, I-131 and I-146; the sulfuric acid salt: compounds I-114 and I-151; the hydrochloride salt: compound I-156; the disodium salt: compound I-115; the mesylate salt: compounds I-106 and I-137; the salt of pamoic acid: compound I-135; and the lysine salt: compound I-216. With continued reference to Tables I and II, certain compounds illustrated as precursor compounds were prepared as one or more salt forms as indicated in the preceding paragraph and in the characterization details provided herein.
[000154] As recognized by one of ordinary skill in the art, the present formulas include other salt forms in addition to those specifically described herein. Similarly, a person of ordinary skill in the art would understand the formulas presently disclosed to encompass solvates such as hydrates.








prodrugs
[000155] Those of skill in the art will appreciate that the 2,4-pyrimidinediamine compounds described herein may include functional groups that can be masked with progroups to create prodrugs. Such prodrugs are usually, but need not be, pharmacologically inactive until converted to their active drug form. Indeed, many of the 2,4-pyrimidinediamine compounds described in this invention include moieties that are hydrolyzable or otherwise cleavable under conditions of use. For example, the ester groups commonly undergo acid catalyzed hydrolysis to produce the precursor carboxylic acid when exposed to acidic conditions in the stomach or base catalyzed hydrolysis when exposed to basic conditions in the intestine or blood. Thus, when administered to a patient orally, 2,4-pyrimidinediamine compounds that include ester moieties can be considered prodrugs of their corresponding carboxylic acids, regardless of whether the ester form is pharmacologically active.
[000156] The mechanism by which the progroup(s) metabolize is not critical and may be caused, for example, by hydrolysis under acidic conditions of the stomach, as described above, and/or by enzymes present in the digestive tract and/or tissues or organs of the body. In fact, the progroup(s) can be selected to metabolize at a particular site within the body. For example, many esters are cleaved under the acidic conditions found in the stomach. Prodrugs designed to chemically cleave in the stomach to the active 2,4-pyrimidine diamine can utilize progroups that include such esters. Alternatively, the progroups can be designed to metabolize in the presence of enzymes such as esterases, amidases, lipolases, and phosphatases, which include ATPases and kinase, etc. Progroups that include bonds capable of metabolizing in vivo are well known and include, By way of example and not limitation, ethers, thioethers, silylethers, silylthioethers, esters, thioesters, carbonates, thiocarbonates, carbamates, thiocarbamates, ureas, thioureas , and carboxamides. In some cases, a “precursor” group that is oxidized by oxidative enzymes such as, for example, liver cytochrome P450, to a metabolizable group, can be selected.
[000157] In prodrugs, any available functional moiety can be masked with a progroup to produce a prodrug. Functional groups within 2,4-pyrimidinediamine compounds that can be masked as progroups for inclusion in a proportion include, but are not limited to, amines (primary and secondary), hydroxyls, sulfanyls (thiols), and carboxyls. A wide variety of progroups, as well as the resulting proportions, suitable for masking functional groups on active 2,4-pyrimidinediamine compounds to produce prodrugs are well known in the art. For example, a hydroxyl functional group can be masked as a sulfonate, ester, or carbonate proportion, which can be hydrolyzed in vivo to provide the hydroxyl group. An amino functional group can be masked as an amide, carbamate, imine, urea, phosphenyl, phosphoryl, or sulphenyl proportion, which can be hydrolyzed in vivo to provide the amino group. A carboxyl group can be masked as a pro-moiety of ester (which includes silyl esters and thioesters), amide, or hydrazide, which can be hydrolyzed in vivo to provide the carboxyl group. Other specific examples of suitable pro-groups and their respective pro-portions will be evident to those of skill in the art. All of these pro-groups, alone or in combination, can be included in pro-drugs.
[000158] In some embodiments of the 2,4-pyrimidinediamine compounds and methods of using the compounds, the progroup(s) may be attached to any available primary or secondary amine, which includes, for example, the 2,4-pyrimidinediamine N2 nitrogen atom, the 2,4-pyrimidinediamine N4 nitrogen atom, and/or a primary or secondary nitrogen atom included in a substituent on the 2,4-pyrimidinediamine .
[000159] As mentioned above, the identity of the progroup is not critical, as long as it can be metabolized under the desired conditions of use, for example, under the acidic conditions found in the stomach and/or by the enzymes found in vivo, to produce a biologically active group, for example, the 2,4-pyrimidinediamines as described herein. Thus, those of skill will appreciate that the progroup can include virtually any known or later discovered hydroxyl, amine or thiol protecting group. Non-limiting examples of suitable protecting groups can be found, for example, in Protective Groups in Organic Synthesis, Greene & Wuts, 2nd Ed., John Wiley & Sons, New York, 1991 (especially on pages 10 to 142 (alcohols) , 277 to 308 (thiols) and 309 to 405 (amines) the disclosure of which is incorporated herein by reference, referred to herein as "Green & Wuts").
[000160] A particularly useful progroup used in exemplary compounds disclosed is -CH2OP(OH)2 as well as esters, mixed acid esters and salts thereof. In some embodiments, the -CH2OP(OH)2 progroup is attached via a nitrogen atom, ring or not, of the precursor molecule. There may be more than one such pro-group. Thus, one embodiment is a compound of formula I,

[000161] or solvate thereof, where A, X, Y, Z1, Z2, R2, R3, R4, R5 and p are as described herein above, and at least each of R1 (when present), R3 and R4 is R50; where R50 is -CH2OP(O)(OR11)2; each R11 is independently for each occurrence H, C1-6 alkyl or monovalent cationic group, or two R11 together with the atoms to which they are attached form a 4- to 8-membered cyclic phosphate group

[000162] where each R55 is independently for each occurrence H, optionally substituted C1-6 alkyl, optionally substituted 3 to 8 membered heteroarylcyclyl, optionally substituted C6-14 aryl, optionally substituted C7-20 arylalkyl, optionally substituted 5 to 14 membered heteroaryl substituted or optionally substituted 6- to 15-membered heteroarylalkyl; z is 0, 1, 2 or 3; or two R11 together represent a divalent inorganic or organic cationic group, where exemplary inorganic divalent cationic groups include those selected from Ba2+, Bi2+, Ca2+, Cu2+, Mg2+, Ni2+, Sr2+, and Zn2+.
[000163] Another embodiment is a compound of formula II

[000164] or solvate thereof, where A, Z1, Z2, R2, R3, R4, R5 and p are as described herein above, and at least each of R1, R3 and R4 is R50; where R50 is -CH2OP(O)(OR11)2; each R11 is independently for each occurrence H, C1-6 alkyl or a monovalent cationic group, or two R11 together with the atoms to which they are attached form a 5- or 6-membered cyclic phosphate group, where -CH2OP(O) (OR11)2 is

[000165] or two R11 together represent a pharmaceutically acceptable divalent cationic group, by way of example including those selected from Ca2+, Mg2+ and Zn2+.
[000166] Another embodiment is a compound of formula III

[000167] or solvate thereof, where A, Z1, Z2, R2, R5 and p are as described herein above, and R50 is -CH2OP(O)(OR11)2; each R11 is independently for each occurrence H, C1-6 alkyl, Li+, K+, HOCH2CH2N(CH3)3+, Na+ or NH4+; or two R11 together represent a divalent cationic group selected from Ca2+, Mg2+ and Zn2+.
[000168] Another embodiment is a compound of formula IV

[000169] or solvate thereof, where A, Z1, Z2, R2, R5 and p are as described herein above, and each R11 is independently for each occurrence H, t-butyl, Li+, HOCH2CH2N(CH3)3+, K+, Na+ or NH4+; or two R11 together represent a divalent cationic group selected from Ca2+, Mg2+ and Zn2+.
[000170] For each of the prodrug embodiments described in the four preceding paragraphs with respect to formulas I, II, III and IV, each of the embodiments with respect to formulas I, IA, IB and IC above if apply. In other words, for each of the embodiments described with respect to formula I, IA, IB and IC above, there is another embodiment where the specific prodrug embodiments described in the previous four paragraphs apply.
[000171] Although not intended to be bound by any particular theory of operation, it is believed that the -CH2OP(O)(OR11)2 progroups, for example, according to formula V, metabolize to active compounds through the intermediary of the corresponding hydroxymethylamine intermediate illustrated below:

[000172] Such hydroxymethylamine compounds, although typically isolatable under controlled conditions, are known to be unstable under physiological conditions and various pH ranges where they hydrolyze in vivo to produce formaldehyde and the active drug substance. Based on this observation, compounds as described herein include hydroxymethyl progroups that can be metabolized in vivo, for example by acidic conditions in the stomach and/or by enzymes present in the digestive tract or other organs and/or tissues or fluids with the body, to produce the active drug substance 2,4-pyrimidinediamine.
[000173] Furthermore, it is expected that the amino and thio analogues of these hydroxymethylamines will be similarly unstable under physiological conditions and will also hydrolyze in vivo to the active 2,4-pyrimidinediamine drug. Accordingly, compounds as described herein include these corresponding primary amino and thiol compounds. Also, the invention includes compounds in which the primary amine, thiol and hydroxy groups are masked with "protecting" groups which are removed under physiological conditions of use to produce the corresponding hydroxymethyl, thiolmethyl and aminomethyl compounds, i.e. with these " protecting groups” these compounds will likewise constitute suitable prodrugs.
[000174] The suitability of any particular progroup for a desired mode of administration can be confirmed in biochemical assays. For example, if a prodrug is to be administered by injection into a particular tissue or organ and the identities of the various enzymes expressed in the tissue or organ are known, the particular prodrug can be tested for metabolism in biochemical assays with ( s) isolated enzyme(s). Alternatively, the particular prodrug can be tested for metabolism to the active 2,4-pyrimidinediamine compound with tissue and/or organ extracts. The use of tissue and/or organ extracts may be of particular convenience when the identity(s) of enzymes expressed in target tissues or organs are unknown or in cases when isolated enzymes are not conveniently available. A person of ordinary skill in the art would be able to easily select progroups having metabolic (such as kinetic) properties suitable for particular applications using such in vitro tests. Specific prodrugs can also be tested for proper metabolism in in vitro animal models.
[000175] Compounds as described herein bearing the pro-group -CH2OP(O)(OR11)2 can be synthesized, for example, as depicted below for compounds of formula IV.

[000176] Typically, the proton in the NH of the oxazolidinone ring can be selectively alkylated with the appropriate phosphonate reagent, where LG is a suitable leaving group to form compounds as described herein, in this case of formula IV. Further description of how to make progroups of the formula -CH2OP(O)(OR11)2 as described herein is specifically disclosed in US Patent No. 7,449,458 entitled "Pyrimidinediamine Pro-drugs and their Uses," the disclosure of which is by hereby incorporated by reference in its entirety.
[000177] A person of skill in the art will appreciate that compounds as described herein may exhibit the phenomena of tautomerism, conformational isomerism, geometric isomerism, and/or optical isomerism. For example, the compounds and prodrugs of the invention may include one or more chiral centers and/or double bonds and as a consequence may exist as stereoisomers, such as double bond isomers (such as geometric isomers), enantiomers, diastereomers, and mixtures thereof, such as racemic mixtures. As another example, compounds as described herein can exist in various tautomeric forms, which include the enol form, the keto form, and mixtures thereof. As the various compound names, formulas, and compound drawings within the specification and claims may represent only one of the possible tautomeric, conformational isomeric, optical isomeric, or geometric isomeric forms, it would be understood that the invention encompasses any of the tautomeric, isomeric forms conformational, optical isomeric, and/or geometric isomeric of the compounds described herein, as well as mixtures of these various different isomeric forms. In cases of limited rotation, for example around the 2,4-pyrimidinediamine core structure, atropisomers are also possible and are also specifically included in the compounds as described herein. It is intended that the compounds encompassed herein are, with the exception of forms of isomerism, chemically stable and isolatable.
[000178] As is understood by a person of skill in the art, certain atoms occur in more than one isotopic form. For example 1 23 hydrogen occurs as protium (H), deuterium (H) and tritium (H), and carbon occurs naturally as three different isotopes, 12C, 13C and 14C. Thus the presently disclosed formulas include compounds having one or more different isotopic forms of certain elements, which include hydrogen and carbon. One of ordinary skill in the art will recognize that any atom at any position in the disclosed compounds can be isotopically enriched, labeled with at least one isotope, or combinations thereof, with any isotope currently known or discovered in the future. Particular examples of isotopes include isotopes of carbon, hydrogen, nitrogen, oxygen, phosphorus, halogens (for example chlorine, fluorine, bromine, and iodine), and combinations thereof. In one embodiment, the presently disclosed compounds are provided in isotopically enriched form. In particular examples, the compounds of formula I are enriched in deuterium relative to protium. In one embodiment, one or more groups attached to the A ring have deuterium in place of H. In another embodiment, an alkyl group in R5 has deuterium in place of H. Deuterium has a natural abundance of about 0.015 %. Consequently, for approximately every 6,500 hydrogen atoms that occur in nature, there is a deuterium atom. Disclosed herein are compounds enriched in deuterium at one or more positions. Thus, the deuterium-containing compounds of the disclosure have deuterium in one or more positions (as the case may be) in an abundance of more than 0.015%.
[000179] In one embodiment, a compound of formula (I), at a position designated as having deuterium, has a minimum isotopic enrichment factor of at least 2000 (30% deuterium incorporation) at each atom designated as deuterium in the compound, or at least 3000 (45% deuterium incorporation).
[000180] In other embodiments, a compound of formula (I) has an isotopic enrichment factor for each planned deuterium atom of at least 3500 (52.5% deuterium incorporation in each planned deuterium atom), at least 4000 ( 60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% of deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99 .5% deuterium incorporation). Pharmaceutical Compositions
[000181] Another embodiment is a pharmaceutical composition that includes a compound as described in any of the embodiments herein. The pharmaceutical compositions described herein can be manufactured by means of conventional mixing, dissolving, granulating, tableting levigating, emulsifying, encapsulating, entrapping or lyophilizing processes. The compositions can be formulated in conventional manner using one or more physiologically acceptable carriers, diluents, excipients, or auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically.
[000182] The 2,4-pyrimidinediamine compound can be formulated into pharmaceutical compositions by itself, or as a hydrate, solvate, N-oxide, or pharmaceutically acceptable salt, as described herein. Typically, such salts are more soluble in aqueous solutions than the corresponding free acids and bases, but salts having lower solubility than the corresponding free acids and bases can also be formed.
[000183] One embodiment is a pharmaceutical formulation that includes a compound of formula I, as described herein, or a prodrug thereof, and at least one pharmaceutically acceptable excipient, diluent, preservative, stabilizer, or mixture thereof.
[000184] The compounds can be provided in a variety of formulations and dosages. The compounds can be provided in a pharmaceutically acceptable form, which includes where the compound can be formulated into pharmaceutical compositions by itself, or in the form of a pharmaceutically acceptable hydrate, solvate, N-oxide, or salt, as described herein. Typically, such salts are more soluble in aqueous solutions than the corresponding free acids and bases, but salts having lower solubility than the corresponding free acids and bases can also be formed. It should be understood that reference to the compound, 2,4-pyrimidinediamine compound, or "active" in formulation debates is also intended to include, where appropriate as known to those of skill in the art, the formulation of prodrugs of the 2- ,4-pyrimidinediamine.
[000185] In one embodiment, the compounds are provided as non-toxic pharmaceutically acceptable salts as mentioned above. Suitable pharmaceutically acceptable salts of the compounds described herein include acid addition salts such as those formed with hydrochloric acid, fumaric acid, p-toluenesulfonic acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, or phosphoric acid. Salts of amine groups can also include quaternary ammonium salts in which the amino nitrogen atom bears a suitable organic group such as an alkyl, alkenyl, alkynyl, or substituted alkyl moiety. In addition, where presently disclosed, the compounds bear an acidic moiety, suitable pharmaceutically acceptable salts thereof may include metal salts such as alkali metal salts, for example, sodium or potassium salts; and alkaline earth metal salts, for example calcium or magnesium salts.
Pharmaceutical compositions for administering the 2,4-pyrimidinediamine compounds may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Pharmaceutical compositions can be, for example, prepared by bringing the active ingredient uniformly and intimately in association with a liquid carrier, a finely divided solid carrier or both, and then, if necessary, shaping the product into the desired formulation. In the pharmaceutical composition the subject active compound is included in an amount sufficient to produce the desired therapeutic effect.
[000187] The 2,4-pyrimidinediamine compounds can be administered orally, parenterally (eg, intramuscular, intraperitoneal, intravenous, ICV, intracystic injection or infusion, subcutaneous injection, or implant), by nasal spray inhalation, vaginal , rectal, sublingual, urethral (eg urethral suppository) or topical administration (eg gel, ointment, cream, aerosol, etc.) and may be formulated, alone or together, into suitable unit dosage formulations containing carriers, pharmaceutically acceptable conventional non-toxic adjuvants, excipients, and vehicles suitable for each via administration. In addition to treating warm-blooded animals such as mice, rats, horses, cattle, sheep, dogs, cats, and monkeys, the compounds described herein can be effective in humans.
[000188] Administration of the compounds described herein, or pharmaceutically acceptable salts thereof, in pure form or in an appropriate pharmaceutical composition, can be carried out via any of the accepted modes of administration or agents to serve similar utilities. Thus, administration can be, for example, oral, nasal, parenteral (intravenous, intramuscular, or subcutaneous), topical, transdermal, intravaginal, intravesical, intracystic, or rectally, in solid, semi-solid, lyophilized powder, or liquid dosage forms, such as, for example, tablets, suppositories, pills, soft and hard elastic gelatin capsules, powders, solutions, suspensions, or aerosols, or the like, preferably in unit dosage forms suitable for simple administration of exact dosages .
[000189] For topical administration, the selective compound(s) or prodrug(s) in JAK can be formulated as solutions, gels, ointments, creams, suspensions, etc., as are well known in technique. Such formulations can be included in a pathway or other transdermal delivery system or formulation, for example, a formulation with ingredients specifically designed to aid in the transport of the compound through the skin and into body tissues.
[000190] Systemic formulations include those intended for administration by injection (eg, subcutaneous, intravenous, intramuscular, intrathecal, or intraperitoneal injection) as well as those intended for transdermal, transmucosal, oral, or pulmonary administration.
[000191] Useful injectable preparations include sterile suspensions, solutions, or emulsions of the active compound(s) in aqueous or oily vehicles. The compositions may also contain formulatory agents, such as suspending, stabilizing, and/or dispersing agents. Formulations for injection may be presented in unit dosage form, for example, in ampoules or in multiple dose containers, and may contain added preservatives.
[000192] Alternatively, the injectable formulation may be provided in powder form for reconstitution with a suitable vehicle, which includes, but is not limited to sterile pyrogen-free water, buffer, and dextrose solution, before use. For this purpose, the active compound(s) can be dried by any technique known in the art, such as lyophilization, and reconstituted before use.
[000193] For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are known in the art.
[000194] For oral administration, pharmaceutical compositions may take the form, for example, of tablets, tablets, or capsules prepared by conventional means with pharmaceutically acceptable excipients such as binding agents (for example, pregelatinized corn starch, polyvinylpyrrolidone , or hydroxypropyl methylcellulose); fillers (for example, lactose, microcrystalline cellulose, or calcium hydrogen phosphate); lubricants (for example, magnesium stearate, talc, or silica); disintegrants (for example potato starch or sodium starch glycolate); or wetting agents (eg, sodium lauryl sulfate). Tablets can be coated by methods well known in the art with, for example, sugars, films, or enteric coatings. Additionally, pharmaceutical compositions containing 2,4-substituted pyrimidinediamine as active ingredient or prodrug thereof in a form suitable for oral use may also include, for example, lozenges, tablets, aqueous or oily suspensions, dispersible powders or granules , emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use can be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions can contain one or more agents including sweetening agents, flavoring agents, coloring agents, and preservative agents in a manner to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient (which includes drug and/or prodrug) in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of tablets. These excipients can be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents (for example corn starch or alginic acid); binding agents (for example starch, gelatin, or acacia); and lubricating agents (eg, magnesium stearate, stearic acid, or talc). The tablets can be left uncoated or they can be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thus provide a prolonged action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate can be used. They can also be coated by the techniques described in U.S. Pat. U.S. No. 4,256,108; 4,166,452; and 4,265,874 to form osmotic therapeutic tablets to control release. The pharmaceutical compositions described herein may also be in the form of oil-in-water emulsions.
[000195] Liquid preparations for oral administration may take the form, for example, of elixirs, solutions, syrups, or suspensions, or they may be presented as a dry product for constitution with water or other suitable vehicles before use. Such liquid preparations can be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (for example, sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (for example, lecithin, or acacia); non-aqueous vehicles (for example, almond oil, oily esters, ethyl alcohol, cremofore®, or fractionated vegetable oils); and preservatives (eg, methyl or propyl p-hydroxybenzoates or sorbic acid). Preparations may also contain buffer salts, preservatives, flavorings, colorings, and sweetening agents as appropriate.
[000196] Preparations for oral administration can be suitably formulated to give controlled release of the active compound, as is well known.
[000197] For oral administration, the compositions may take the form of tablets or tablets formulated in conventional manner.
[000198] For nasal administration or administration by inhalation or insufflation, the active compound(s) or prodrug(s) can be conveniently released in the form of a dry powder (alone, as a mixture, eg in a dry mix with lactose, or as a mixed component particle, eg mixed with phospholipids such as phosphatidylcholine) form a dry powder inhaler or as an aerosol spray from pressurized packs or a nebulizer with the use of a suitable propellant (for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, fluorocarbons, carbon dioxide, or other suitable gas). In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges for use in an inhaler or insufflator (for example, capsules and cartridges that include gelatin) may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
[000199] Prior to use in a dry powder or suspension formulation, the prodrug product typically is micronized to a size suitable for delivery by inhalation (typically less than about 5 microns). This can be achieved as is known to those skilled in the art by a suitable method such as spiral jet milling, fluid bed jet milling, supercritical fluid processing, spray drying and the like.
[000200] The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. The 2,4-pyrimidinediamine Compounds can also be administered in the form of suppositories for the rectal or urethral administration of the drug. For the rectal and vaginal routes of administration, the active compound(s) may be formulated as solutions (for retention enemas), suppositories, or ointments containing conventional suppository bases such as cocoa butter or other glycerides. In particular embodiments, the compounds may be formulated as urethral suppositories, for example, for use in the treatment of fertility conditions, particularly in men (for example, for the treatment of testicular dysfunction).
[000201] The presently disclosed 2,4-pyrimidinediamine compounds can be used for the manufacture of a composition or medicine, which includes medicines suitable for topical administration. Accordingly, specifically contemplated are methods for making compositions which include the 2,4-pyrimidinediamine compound in a form that is suitable for topical administration.
[000202] For topical use, creams, ointments, jellies, gels, solutions, suspensions, etc., containing the 2,4-pyrimidinediamine compounds can be used. In certain embodiments, the 2,4-pyrimidinediamine compounds can be formulated for topical administration with polyethylene glycol (PEG). These formulations may optionally include additional pharmaceutically acceptable ingredients such as diluents, stabilizers, and/or adjuvants. In particular embodiments, topical formulations are formulated for the treatment of allergic conditions and/or skin conditions including psoriasis, contact dermatitis, and atopic dermatitis, among others described herein.
[000203] As those skilled in the art will recognize, the formulation of 2,4-pyrimidinediamine compounds, the amount of the formulation released, and the duration of administration of a single dose depends on the type of inhalation device used as well as other factors. For some aerosol delivery systems, such as nebulizers, the frequency of administration and the length of time the system is activated will depend primarily on the concentration of 2,4-pyrimidinediamine compounds in the aerosol. For example, shorter periods of administration can be used at higher concentrations of 2,4-pyrimidinediamine compounds in the nebulizer solution. Devices such as metered dose inhalers can produce higher aerosol concentrations and can be operated for shorter periods to deliver the desired amount of 2,4-pyrimidinediamine compounds in some embodiments. Devices such as dry powder inhalers release active agent until a given load of agent is expelled from the device. In this type of inhaler, the amount of 2,4-pyrimidinediamine compounds in a given amount of powder determines the dose delivered in a single administration. The 2,4-pyrimidinediamine formulation is selected to produce the desired particle size in the chosen inhaler device.
[000204] Included among the devices that can be used to administer particular examples of the 2,4-pyrimidinediamine compounds are those well known in the art, such as metered dose inhalers, liquid nebulizers, dry powder inhalers, sprays, thermal vaporizers, and others. Another technology suitable for the delivery of particular 2,4-pyrimidinediamine compounds includes electrohydrodynamic aerosols.
[000205] Furthermore, the inhalation device is preferably practical, in the sense of being easy to use, small enough to carry conveniently, capable of delivering multiple doses, and durable. Some specific examples of commercially available inhalation devices are Turbohaler (Astra, Wilmington, DE), Rotahaler (Glaxo, Research Triangle Park, NC), Diskus (Glaxo, Research Triangle Park, NC), the Ultravent nebulizer (Mallinckrodt), the nebulizer Acorn II (Marquest Medical Products, Totowa, NJ), the Ventolin metered dose inhaler (Glaxo, Research Triangle Park, NC), and others. In one embodiment, the 2,4-pyrimidinediamine compounds can be delivered by a dry powder inhaler or a spray.
[000206] Formulations of 2,4-pyrimidinediamine compounds for administering a dry powder inhaler may typically include a finely divided dry powder containing 2,4-pyrimidinediamine compounds, but the powder may also include a bulking agent, buffer, filler, excipient, another additive, or the like. In one aspect, the hydroxynaphthoate salts such as 1-hydroxy- or 3-hydroxy-2-naphthalenecarboxylate of the present 2,4-pyrimidinediamines have low hygroscopicity as compared to the corresponding free base form and have excellent compatibility with excipients, stability , micronize well and other pharmaceutical properties. The 1-hydroxy-2-naphthalenecarboxylate salts may be referred to as "xinafoate salts." Disclosed herein are xinafoate salts of 2,4-pyrimidinediamine compounds described herein. The xinafoate salts of 2,4-pyrimidinediamines described herein can be administered alone but can also be administered as a formulation in association with one or more pharmaceutically acceptable diluents, excipients or the like. Dry powder formulations of the xinafoic acid salt forms of the 2,4-pyrimidinediamine compounds described herein are one embodiment. Such dry powder formulations with or without a carrier or other additive are suitable for administration via inhalation. Additives can be included in a dry powder formulation of 2,4-pyrimidinediamine compounds, for example, to dilute the powder to facilitate release from a dry powder inhaler, to facilitate formulation processing, to provide properties of powders advantageous to the formulation, to facilitate dispersion of the powder from the inhalation device, to stabilize the formulation (e.g., antioxidants or buffers), to provide flavor to the formulation, or the like. Typical additives include mono-, di-, and polysaccharides; sugar alcohols and other polyols, such as, for example, lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch, or combinations thereof; surfactants such as sorbitols, diphosphatidyl choline, or lecithin; and others.
[000207] For example, a dry powder formulation can be manufactured in various ways, using conventional techniques, such as described in any of the publications mentioned above and expressly incorporated herein by reference, and, for example, Baker, et al., Pat. No. 5,700,904, the entire disclosure of which is expressly incorporated herein by reference. Particles in the appropriate size range for maximum deposition in the lower respiratory tract can be manufactured by micronizing, milling, or the like. A liquid formulation can be manufactured by dissolving one or more of the presently disclosed 2,4-pyrimidinediamine compounds in a suitable solvent, such as water, at an appropriate pH, which includes buffers or other excipients.
[000208] For ocular administration, the 2,4-pyrimidinediamine compound(s) or prodrug(s) may be formulated as a suitable solution, emulsion, suspension, etc. for administration to the eye. Administration to the eye is generally via topical eye exposure to the formulation, but also includes injection into the eye if necessary. A variety of vehicles suitable for delivering compounds to the eye are known in the art. Specific non-limiting examples are described in U.S. Patent No. 6,261,547; US Patent No. 6,197,934, US Patent No. 6,056,950, US Patent No. 5,800,807, US Patent No. 5,776,445, US Patent No. 5,698,219, US Patent No. 5,521,222, US Patent No. 5,403,841, Patent US Patent No. 5,077,033 , US Patent No. 4,882,150, and US Patent No. 4,738,851.
[000209] Typically formulations for ocular administration contain a pharmaceutically effective amount of a 2,4-pyrimidinediamine compound disclosed herein, such as from about 0.0001% to about 1.0% by weight (w/w) . In certain formulations, the pharmaceutically effective amount of the compound is from 0.0003% to about 0.1% (w/w), such as from about 0.003% to about 0.5% (w/w), or from about 0.01% to about 0.03% (w/w).
[000210] In certain examples an ophthalmic composition containing a presently disclosed 2,4-pyrimidinediamine compound for ocular administration includes a tonicity agent, a buffer, or both. In certain examples of ophthalmic compositions the tonicity agent is a simple carbohydrate or sugar alcohol. As is known to those of skill in the art, tonicity agents can be used in the present compositions to adjust the tonicity of the composition, preferably to that of normal tears. Examples of suitable tonicity agents include, without limitation, sodium chloride, potassium chloride, magnesium chloride, calcium chloride, carbohydrates such as dextrose, fructose, galactose, polyols such as sugar alcohols, which includes. example, mannitol, sorbitol, xylitol, lactitol, isomalt, maltitol and combinations thereof. Compositions containing a buffer contain, in some examples, a phosphate, citrate, or both.
[000211] In one aspect, compositions for the ocular administration of the presently disclosed 2,4-pyrimidinediamine compounds optionally contain a surfactant, a stabilizing polymer, or both. Surfactants are used in certain compositions to facilitate the release of higher concentrations of the 2,4-pyrimidinediamine compound that is administered. Such surfactants can function to solubilize the compound. Exemplary surfactants include polysorbate, poloxamer, polyosil 40 stearate, polyoxyl castor oil, tyloxapol, triton and sorbitan monolaurate. In certain embodiments the surfactant is selected from Triton X114, tyloxapol and combinations thereof. In yet another embodiment of compositions for ocular administration, the stabilizing polymer is 974p carbomer.
[000212] For extended release, the 2,4-pyrimidinediamine compound(s) or prodrug(s) may be formulated as a depot preparation for administration by implantation or intramuscular injection . The active ingredient may be formulated with suitable polymeric or hydrophobic materials (eg as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives (eg as a sparingly soluble salt). Alternatively, transdermal delivery systems manufactured as an adhesive disc or patch that slowly release the active compound(s) for percutaneous absorption can be used. For this purpose, permeation enhancers can be used to facilitate transdermal penetration of the active compound(s). suitable transdermal patches are described, for example, in US Patent No. 5,407,713, US Patent No 5,352,456, US Patent No 5,332,213, US Patent No 5,336,168, US Patent No 5,290,561, US Patent No 5,254 ,346, US Patent No. 5,164,189, US Patent No. 5,163,899, US Patent No. 5,088,977, US Patent No. 5,087,240, US Patent No. 5,008,110, and US Patent No. 4,921,475.
[000213] Alternatively, other pharmaceutical delivery systems can be used. Liposomes and emulsions are well known examples of delivery vehicles that can be used to deliver active compound(s) or prodrug(s). Certain organic solvents such as dimethyl sulfoxide (DMSO) can also be used, although usually at the expense of enormous toxicity.
[000214] Pharmaceutical compositions, if desired, may be presented in a pack or dispensing device which may contain one or more unit dosage forms containing the active compound(s). The packaging, for example, can include metallic or plastic foil, such as a blister pack. The pack or dispensing device may be accompanied by instructions for administration.
[000215] Another embodiment is a kit which includes a compound, prodrug or pharmaceutical composition as described in any of the above embodiments. Kit embodiments are described in more detail below. Methods of use
The present invention provides 2,4-pyrimidinediamine compounds, prodrugs and pharmaceutical compositions thereof, as described herein, for use in therapy for the conditions described herein. The present invention further provides the use of the compounds of the present invention in the manufacture of a medicament for the treatment of conditions where JAK pathway targeting or inhibition of JAK kinases, particularly JAK3, is therapeutically useful. These include conditions where lymphocyte, macrophage, or mastoid function is involved. Conditions where targeting the JAK pathway or inhibiting JAK kinases, particularly JAK3, are therapeutically useful include leukemia, lymphoma, transplant rejection (e.g. pancreatic islet transplant rejection), bone marrow transplant applications (by example, graft-versus-host disease), autoimmune diseases (e.g., rheumatoid arthritis, psoriasis, and the like), inflammation (e.g., asthma, etc.) and other conditions as described in greater detail herein.
[000217] In another embodiment, the methods can be practiced as a therapeutic method for the treatment of the conditions described herein. Thus, in a specific embodiment, the 2,4-pyrimidinediamine compounds (and the various forms described herein, which include pharmaceutical formulations that include the compounds (in the various forms)) can be used to treat the conditions described herein in patients animals, which include humans. The methods generally include administering to the patient an amount of a compound described herein, or a salt, prodrug, hydrate, or N-oxide thereof, effective to treat the condition. In one embodiment, the patient is a non-human mammal, which includes, but is not limited to, a bovine, equine, feline, canine, rodent, or primate. In another embodiment, the patient is a human being.
[000218] As mentioned above, numerous conditions can be treated using the 2,4-substituted pyrimidinediamine compounds, prodrugs thereof, and methods of treatment as described herein. As used herein, "treating" or "treating" a disease in a patient refers to (1) preventing the disease from occurring in a patient who is predisposed to or does not yet demonstrate symptoms of the disease; (2) inhibit the disease or arrest its development; or (3) ameliorate or cause disease regression. As is well understood in the art, "treatment" is a method of obtaining beneficial or desired results, which includes clinical results. For purposes of this invention, beneficial or desired results may include one or more, but are not limited to, alleviation or amelioration of one or more symptoms, lessening of a condition, which includes a disease, stabilized state (i.e., does not worsen) of a condition, which includes disease, prevent the spread of disease, delay or decrease the condition, which includes disease, progression, improvement or relief of the condition, which includes disease, condition, and remission (whether partial or total), whether detectable or undetectable. Preferred are compounds that are relatively potent compared to the class as a whole and can be administered in low doses, preferably but not necessarily locally, thus minimizing systemic adverse effects.
[000219] The compounds described herein are potent and selective inhibitors of JAK kinases and are particularly selective for JAK3-containing cytokine signaling pathways. As a consequence of this activity, the compounds can be used in a variety of in vitro, in vivo, and ex vivo contexts to regulate or inhibit JAK kinase activity, signaling cascades in which JAK kinases play a role, and biological responses effected by such signaling cascades. For example, in one embodiment, the compounds can be used to inhibit JAK kinase, in vitro or in vivo, in virtually any cell type that expresses JAK kinase, such as in hematopoietic cells in which, for example, JAK3 is predominantly expressed. These can also be used to regulate the signal transduction cascades in which JAK kinases, particularly JAK3, play a role. Such JAK-dependent signal transduction cascades include, but are not limited to, cytokine receptor signaling cascades that involve the common gamma chain, such as, for example, the IL-4, IL-7 receptor signaling cascades , IL-5, IL-9, IL-15 and IL-21, or IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21. The compounds can also be used in vitro or in vivo to regulate, and in particular to inhibit, cellular or biological responses affected by such JAK-dependent signal transduction cascades. Such cellular or biological responses include, but are not limited to, up-regulation of IL-4/CD23 branches and IL-2 mediated T cell proliferation. Importantly, the compounds can be used to inhibit JAK kinases in vivo as a therapeutic method for the treatment or prevention of diseases mediated, in whole or in part, by a JAK kinase activity (referred to herein as "diseases mediated by JAK kinase"). Non-limiting examples of JAK kinase mediated diseases that can be treated or prevented with the presently disclosed compounds include, but are not limited to the following: allergies; asthma; autoimmune diseases, which include systemic autoimmune disorders, transplant rejection (eg, kidney, heart, lung, liver, pancreas, skin; host versus graft reaction (HVGR), and graft versus host reaction (GVHR)), rheumatoid arthritis , and amyotrophic lateral sclerosis; T cell-mediated autoimmune diseases such as multiple sclerosis, psoriasis, and Sjogren's syndrome; Type II inflammatory diseases such as vascular inflammation (which includes vasculitis, arteritis, atherosclerosis, and coronary artery disease); central nervous system diseases such as stroke; pulmonary diseases such as obliterative bronchitis and primary pulmonary hypertension; solid, delayed Type IV hypersensitivity reactions; and hematologic malignancies such as leukemia and lymphomas.
[000220] In addition to the disorders listed above, the disclosed compounds are particularly useful for the treatment of obstructive, restrictive or inflammatory airway diseases of any type, etiology, or pathogenesis which are, in particular an obstructive, restrictive or inflammatory, including, as mentioned above, asthma, in particular atopic asthma, allergic asthma, non-atopic asthma, bronchial asthma, non-allergic asthma, emphysematous asthma, exercise-induced asthma, emotion-induced asthma, extrinsic asthma caused by environmental factors, asthma infectious disease associated with bacterial, fungal, protozoa, and/or viral infection, bronchiolitis, cough-variant asthma, drug-induced asthma, and the like. The presently disclosed compounds are also particularly useful in the treatment of rhinitis or sinusitis of different etiologies, including without limitation, seasonal allergic rhinitis, perennial allergic rhinitis, vasomotor rhinitis, sinusitis, including acute, chronic, ethymoid, frontal maxillary or sphenoid sinusitis. The disclosed compounds are also useful in the treatment of chronic obstructive pulmonary disease (COPD), chronic obstructive lung disease (COLD), chronic obstructive airway disease (COAD) or airway obstruction, including, without limitation, chronic bronchitis, emphysema pulmonary, bronchiectasis, cystic fibrosis, bronchiolitis obliterans. In addition, the disclosed compounds can be used to treat bronchitis, including in particular, acute bronchitis, acute laryngotracheal bronchitis, chronic bronchitis, dry bronchitis, productive bronchitis, infectious asthmatic bronchitis, staphylococcal bronchitis or bronchococcal and streptococcal bronchitis.
[000221] An embodiment is a method as described herein used with a compound according to formula I, or in a more specific embodiment, a compound according to formulas IA, IB, IC, II, III or IV in an even more specific embodiment, a species described herein. For brevity, the methods described below refer to a compound of the formula I, but corresponding methods according to the various subgenera and species of compound and composition are also intended to be included.
[000222] One embodiment is a method of inhibiting a JAK kinase activity, which includes contacting the JAK kinase with an amount of a compound, effective to inhibit a JAK kinase activity, of formula I:
Where
[000223] X and Y are each independently O, S, S(O), SO2 or NR1;
[000224] each R1 is independently for each occurrence H, optionally substituted C1-6 alkyl, C(O)-C1-6 alkyl, CO2-C1-6 alkyl or R50;
[000225] each R50 is -C(R9)2-A-R10, where A is O or S; each R9 is independently for each occurrence H, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl, or optionally substituted C7-16 arylalkyl; or alternatively, two R 9 , together with the carbon to which they are attached, form an optionally substituted C 3-8 cycloalkyl group or an optionally substituted 3- to 8-membered heteroarylcyclyl group; R10 is Ra, -P(O)(OR11)2, -P(O)(OR11)N(R12)2 or -P(O)(N(R12)2)2; each R11 is independently for each occurrence Ra or a monovalent cationic group; either two R11 together with the atoms to which they are attached form a 4- to 8-membered cyclic phosphate group, or two R11 together represent a divalent cationic group; each R12 is independently for each occurrence Rc or -C1-3-alkylN(Rc)2; or two R12, each on nitrogens separate from -P(O)(N(R12)2)2, together with the atoms to which they are attached, form a 4- to 8-membered cyclic phosphonic acid bisamide group; or an R12 together with R11, of the group -P(O)(OR11)N(R12)2, together with the atoms to which they are attached, form a 4- to 8-membered cyclic phosphonamidate group;
[000226] ring A is a C6-10 aryl or a 5- to 10-membered heteroaryl;
[000227] each R2 is independently for each occurrence H, Re, Rb, Re substituted with one or more of the same or different Ra and/or Rb, -ORe substituted with one or more of the same or different Ra and/or Rb, - SRe substituted with one or more of the same or different Ra and/or Rb, -C(O)Re substituted with one or more of the same or different Ra and/or Rb, -N(Ra)Re where Re is substituted with one or more of the same or different Ra and/or Rb, - S(O)2Re replaced with one or more of the same or different Ra and/or Rb, - N(Ra)-S(O)2Re where Re is replaced with one or more of the same or different Ra and/or Rb, -B(ORa)2, -B(N(Rc)2)2, -(C(Ra)2)m-Rb, -O-(C(Ra)2 )m-Rb, -abba ab S-(C(R)2)mR, -O-(C(R)2)mR, -N(R)-(C(R)2)mR, -O -(CH2)m-bb aabaa CH((CH2)mR)R, -C(O)N(R)-(C(R)2)mR, -O-(C(R)2)mC(O) N(R )- ab ab aaab (C(R )2)mR , -N((C(R )2)mR )2, -S-(C(R )2)mC(O)N(R )- (C(R )2)mR , - aaabaa ab N(R )-C(O)-N(R )-(C(R )2)mR , -N(R )-C(O)-(C( R )2)mC(R)(R)2 or -N(Ra)-(C(Ra)2)mC(O)-N( Ra)-(C(Ra)2)m-Rb;
[000228] each Ra is independently for each occurrence H, deuterium, C1-6 alkyl, C3-8 cycloalkyl, C4-11 cycloalkylalkyl, C6-10 aryl, C7-16 arylalkyl, 2 to 6-membered heteroalkyl, 3 to 3-membered heteroalicyclyl 10-membered, 4- to 11-membered heteroalicyclylalkyl, 5- to 15-membered heteroaryl, or 6- to 16-membered heteroarylalkyl;
[000229] each Rb is independently for each occurrence =O, -ORa, -O-(C(Ra)2)m-ORa, halo-C1-3 alkyloxy, =S, -SRa, =NRa, =NORa, - N(Rc)2, halo, -CF3, -CN, -NC, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)Ra, -S(O)2Ra, - SO3Ra, -S(O)N(Rc)2, -S(O)2N(Rc)2, -OS(O)Ra, -OS(O)2Ra, -OSO3Ra, - OS(O)2N(Rc) 2, -C(O)Ra, -CO2Ra, -C(O)N(Rc)2, -C(NRa)-N(Rc)2, -C(NOH)-Ra, -C(NOH)-N (Rc)2, -OC(O)Ra, -OC(O)ORa, -OC(O)N(Rc)2, -OC(NH)-N(Rc)2, -ac aaaaa OC(NR )- N(R )2, -N(R )-S(O)2H, -[N(R )C(O)]nR , -[N(R )C(O)]nOR , - [N(Ra) C(O)]nN(Rc)2 or -[N(Ra)C(NRa)]nN(Rc)2;
[000230] each Rc is independently for each occurrence Ra, or, alternatively, two Rc are taken together with the nitrogen atom to which they are attached to form a 3- to 10-membered heteroalicyclyl or a 5- to 10-membered heteroaryl that can optionally including one or more of the same or different additional heteroatoms and which is optionally substituted with one or more of the same or different Ra and/or Rd groups;
[000231] each Rd is =O, -ORa, C1-3 haloalkyloxy, C1-6 alkyl, =S, -SRa, =NRa, =NORa, -N(Ra)2, halo, -CF3, -CN, - NC, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)Ra, -S(O2)Ra, -SO3Ra, -S(O)N(Ra)2, -S (O)2N(Ra)2, -OS(O)Ra, -OS(O)2Ra, -OSO3Ra, -OS(O)2N(Ra)2, -C(O)Ra, -CO2Ra, -C( O)N(Ra)2, -C(NRa)N(Ra)2, -C(NOH)Ra, -C(NOH)N(Ra)2, -OCO2Ra, -OC(O)N(Ra)2 , - aa aaaaaa OC(NR )N(R )2, -[N(R )C(O)]nR , -(C(R )2)n-OR , -N(R )-S(O)2R , -C(O)-C1-6 haloalkyl, -S(O)2C1-6 haloalkyl, -OC(O)Ra, -O(C(Ra)2)m-ORa, - aa a aa to S( C(R)2)m-OR, -N(R)C1-6 haloalkyl, -P(O)(OR)2, -N(R)-(C(R)2)m-OR, -[N (Ra)C(O)]nORa, -[N(Ra)C(O)]nN(Ra)2, -[N(Ra)C(NRa)]nN(Ra)2 or - N(Ra)C (O)haloC1-6alkyl; two Rd, taken together with the atom or atoms to which they are attached, combine to form a partially or fully saturated 3- to 10-membered mono or bicyclic ring, optionally containing one or more heteroatoms and optionally substituted with one or more Ra;
[000232] each Re is independently for each occurrence C1-6 alkyl, C3-8 cycloalkyl, C4-11 cycloalkylalkyl, C6-10 aryl, C7-16 arylalkyl, 2 to 6 membered heteroalkyl, 3 to 10 membered heteroalicyclyl, heteroalicyclylalkyl 4- to 11-membered, 5- to 15-membered heteroaryl, or 6- to 16-membered heteroarylalkyl;
[000233] p is 0, 1, 2, 3 or 4;
[000234] each m is 1, 2 or 3;
[000235] each n is 0, 1, 2 or 3;
[000236] two R2 groups, taken together with the atom or atoms to which they are attached, combine to form a partially or fully saturated 4- to 10-membered mono or bicyclic ring, optionally containing one or more heteroatoms and optionally substituted with one or more Ra and/or Rb;
[000237] Z1 and Z2 are each independently CH, CR2 or N;
[000238] R3 is H, optionally substituted C1-6 alkyl or R50;
[000239] R4 is H, optionally substituted C1-6 alkyl or R50; and
[000240] R5 is H, halo, -CN, optionally substituted C1-6 alkyl, alkynyl, hydroxy, optionally substituted C1-6 alkoxy, nitro, -N(Ra)2, -C(O)N(Ra)2, -CO2Ra or -C(O)Ra.
[000241] In another embodiment, a method of inhibiting a JAK kinase activity, which includes contacting the JAK kinase with an amount of a compound effective to inhibit a JAK kinase activity, where the compound conforms to formula I , as described here. In certain embodiments of the methods described herein, the method is performed in vivo.
[000242] In another embodiment, this invention provides a method of inhibiting a JAK kinase activity, which includes contacting in vitro a JAK3 kinase with an amount of a compound effective to inhibit a JAK kinase activity, where the compound is of according to formula I as described herein.
[000243] In a specific embodiment, the compounds can be used to treat and/or prevent rejection in organ and/or tissue transplant recipients (i.e., treat and/or prevent halograft rejection). Halografts can be rejected through a cell-mediated or humoral immune reaction of the recipient's antigens against the transplant (histocompatibility) present in the membranes of the donor's cells. The strongest antigens are controlled by a complex of genetic loci called human leukocyte group A antigens (HLA). Together with ABO blood group antigens, they are the main detectable transplant antigens in humans.
[000244] Rejection following transplantation in general can be divided into three categories: hyperacute, which occurs hours to days following transplantation; acute, which occurs days to months after transplantation; and chronic, which occurs months to years after transplantation.
[000245] Hyperacute rejection is mainly caused by the production of host antibodies that attack the graft tissue. In a hyperacute rejection reaction, antibodies are seen in vascular transplantation shortly after transplantation. Immediately thereafter, vascular clots occur, leading to ischemia, eventual necrosis, and death. Graft infarction is not responsive to known immunosuppressive therapies. Because HLA antigens can be identified in vitro, pre-transplant screening is used to significantly reduce hyperacute rejection. As a consequence of this screening, hyperacute rejection is relatively uncommon these days.
[000246] Acute rejection is considered to be mediated by the accumulation of antigen-specific cells in the graft tissue. The T cell-mediated immune reaction against these antigens (ie, HVGR or GVHR) is the main mechanism of acute rejection. The accumulation of these cells leads to damage to the graft tissue. It is believed that both CD4+ helper T cells and CD8+ cytotoxic T cells are involved in the process and that the antigen is presented by dendritic cells of the donor and host. CD4+ helper T cells help to recruit other effector cells, such as macrophages and eosinophils, into the engraftment. Accession of the T cell activation signal transduction cascades (eg, the CD28, CD40L, and CD2 cascades) is also involved.
[000247] Acute cell-mediated rejection can be reversed in many cases by intensification of immunotherapy. After successful reversal, severely damaged elements of the graft heal from fibrosis and the rest of the graft appears normal. After resolution of acute rejection, dosages of immunosuppressive drugs can be reduced to very low levels.
[000248] Chronic rejection, which is a particular problem in kidney transplants, often progresses insidiously despite augmented immunosuppressive therapy. This is considered to be due in large part to cell-mediated Type IV hypersensitivity. The pathological profile differs from that of acute rejection. The arterial endothelium is primarily involved with extensive proliferation that can gradually occlude the vessel lumen, leading to ischemia, fibrosis, a thickened intima, and atherosclerotic changes. Chronic rejection is primarily due to a progressive obliteration of the graft vasculature and resembles a slow, vasculitic process.
[000249] In Type IV hypersensitivity, cytotoxic CD8 T cells and CD4 helper T cells recognize intracellular or extracellular synthesized antigen when it is complexed, respectively, with Class I or Class II MHC molecules. Macrophages function as antigen-presenting cells and release IL-1, which promotes the proliferation of helper T cells. Helper T cells release interferon gamma and IL-2, which together regulate macrophage activation-mediated hyperactivity reactions and T cell-mediated immunity. In the case of organ transplantation, cytotoxic T cells destroy the graft cells on contact.
[000250] Since JAK kinases play a critical role in T cell activation, the 2,4-pyrimidinediamine compounds described herein can be used to treat and/or prevent many aspects of transplant rejection, and are particularly useful in treating and/or preventing rejection reactions that are mediated, at least in part, by T cells, such as HVGR or GVHR. The 2,4-pyrimidinediamine compounds can also be used to treat and/or prevent chronic rejection in transplant recipients and, in particular, in renal transplant recipients. The compound can also be administered to a tissue or an organ prior to transplanting the tissue or organ into the transplant recipient.
[000251] In another embodiment, this invention provides a method of treating a T cell-mediated autoimmune disease, which includes administering to a patient suffering from such an autoimmune disease an amount of a compound effective to treat the autoimmune disease where the compound is in accordance with formula I as described herein. In certain embodiments of the methods the autoimmune disease is multiple sclerosis (MS), psoriasis, or Sjogran's syndrome.
Therapy using the 2,4-pyrimidinediamine compounds described herein can be applied alone, or it can be applied in combination with or adjunctively with other common immunosuppressive therapies, such as, for example, the following: mercaptopurine; corticosteroids such as prednisone; methylprednisolone and prednisolone; alkylating agents such as cyclophosphamide; calcineurin inhibitors such as cyclosporine, sirolimus, and tacrolimus; inosine dehydrogenase monophosphate (IMPDH) inhibitors such as mycophenolate, mycophenolate mofetil, and azathioprine; and agents designed to suppress cellular immunity while leaving the recipient's humoral immune response intact, which include various antibodies (eg, anti-lymphocyte globulin (ALG), anti-thymocyte globulin (ATG), anti-T cell monoclonal antibodies (OKT3)) and irradiation . These various agents can be used according to their standard or common dosages, as specified in the prescribed information accompanying the commercially available forms of the drugs (see also: the prescribed information in the 2006 Edition of the Physician's Desk Reference), the disclosures of which are hereby incorporated by reference. Azathioprine is currently available from Salix Pharmaceuticals, Inc., under the tradename AZASAN; mercaptopurine is currently available from Gate Pharmaceuticals, Inc. under the tradename PURINETOL; prednisone and prednisolone are currently available from Roxane Laboratories, Inc.; Methyl prednisolone is currently available from Pfizer; sirolimus (rapamycin) is currently available from Wyet-Ayerst under the tradename RAPAMUNE; tacrolimus is currently available from Fujisawa under the tradename PROGRAF; cyclosporin is currently available from Novartis under the tradename SANDIMMUNE and from Abbott under the tradename GENGRAF; IMPDH inhibitors such as mycophenolate mofetil and mycophenolic acid are currently available from Roche under the tradename CELLCEPT and from Novartis under the tradename MYFORTIC; azathioprine is currently available from Glaxo Smith Kline under the tradename IMURAN; and antibodies are currently available from Ortho Biotech under the tradename ORTHOCLONE, from Novartis under the tradename SIMULECT (basiliximab), and from Roche under the tradename ZENAPAX (daclizumab).
[000253] In another embodiment, the 2,4-pyrimidinediamine compounds can be administered in combination or adjunct to an inhibitor of a Syk kinase. Syk kinase is a tyrosine kinase known to play a critical role in FCY receptor signaling, as well as other signaling cascades, such as those involving B cell receptor signaling (Tumer et al., (2000), Immunology Today 21 :148-154) and beta(1), beta(2), and beta(3) integrins on neutrophils ( Mocsavi et al., (2002), Immunity 16: 547-558). For example, Syk kinase plays a pivotal role in high-affinity IgE receptor signaling in mastoids that leads to activation and subsequent release of multiple chemical mediators that trigger allergic attacks. However, unlike JAK kinases, which help regulate pathways involved in delayed or cell-mediated Type IV hypersensitivity reactions, Syk kinase helps regulate pathways involved in immediate IgE-mediated Type I hypersensitivity reactions. Certain compounds that affect the Syk pathway may or may not affect JAK pathways as well.
Suitable Syk inhibitor compounds are described, for example, in Serial No. 10/355,543 filed January 31, 2003 (publication no. 2004/0029902); WO 03/063794; Serial No. 10/631,029 filed on July 29, 2003 (publication No. 2007/0060603); WO 2004/014382; Serial No. 10/903,263 filed July 30, 2004 (publication no. 2005/0234049); PCT/US2004/24716 filed July 30, 2004 (WO005/016893); Serial No. 10/903,870 filed July 30, 2004 (publication no. 2005/0209224); PCT/US2004/24920 filed July 30, 2004; Serial No. 60/630,808 filed November 24, 2004; Serial No. 60/645,424 filed January 19, 2005; and Serial No. 60/654,620, filed February 18, 2005, the disclosures of which are incorporated herein by reference. The 2,4-pyrimidinediamine described herein and Syk inhibitor compounds can be used alone or in combination with one or more conventional transplant rejection treatments as described above.
[000255] In a specific embodiment, 2,4-pyrimidinediamine compounds can be used to treat or prevent these diseases in patients who are initially unresponsive (resistant) to or who become unresponsive to treatment with an inhibitor compound of Syk or one of the other current treatments for the particular disease. The 2,4-pyrimidinediamine compounds can also be used in combination with Syk inhibitor compounds in patients who are resistant or unresponsive to the Syk compound. Suitable Syk inhibitor compounds with which the 2,4-pyrimidinediamine compounds can be administered are provided above.
[000256] In another embodiment, this invention provides a method of treating a T cell-mediated autoimmune disease, which includes administering to a patient suffering from such an autoimmune disease an amount of a compound according to formula I, in combination with or adjunctively with a compound that inhibits Syk kinase with an IC50 of at least 10 µM, effective to treat the autoimmune disease.
[000257] In another embodiment, this invention provides a method of treating acute or chronic allograft transplant rejection in a transplant recipient, which includes administering to the transplant recipient an amount of a compound in accordance with the formula I effective in treating or preventing rejection. in another embodiment, the compound is administered to a tissue or an organ prior to, or concurrent with, transplanting the tissue or organ in the transplant recipient. In another embodiment, the compound is administered to the tissue or organ and the patient. In a specific embodiment allograft transplant rejection is mediated by HVGR or GVHR. In another embodiment, the transplanted halograft organ is a kidney, heart, liver, or lung. In another embodiment, where the transplanted halograft organ is a kidney, heart, liver, or lung, the compound is administered in combination with or adjunct to another immunosuppressant. In a more specific embodiment, the immunosuppressant is cyclosporine, tacrolimus, sirolimus, an IMPDH inhibitor, mycophenolate, mycophanolate mofetil, an anti-T cell antibody or OKT3.
[000258] The 2,4-pyrimidinediamine compounds described herein are cytokine moderators of IL-4 signaling. As a consequence, 2,4-pyrimidinediamine compounds can decrease the response of Type I hypersensitivity reactions. Thus, in a specific embodiment, 2,4-pyrimidinediamine compounds can be used to treat such reactions and therefore , diseases associated with, mediated by, or caused by such hypersensitivity reactions (eg, allergies), prophylactically. For example, an allergy sufferer may take one or more of the selective JAK compounds described herein prior to expected exposure to the allergens to delay the onset or progression of, or completely eliminate, an allergic response.
[000259] When used to treat or prevent such diseases, the 2,4-pyrimidinediamine Compounds can be administered alone, as mixtures of one or more 2,4-pyrimidinediamine compounds, or in mixture or combination with other agents useful to treat such diseases and/or the symptoms associated with such diseases. The 2,4-pyrimidinediamine Compounds can also be administered in admixture or in combination with agents useful to treat other disorders or ailments, such as steroids, membrane stabilizers, 5-lipoxygenase (5LO) inhibitors, leukotriene synthesis, and receptor inhibitors , IgE isotype switching or IgE synthesis inhibitors, IgG isotype switching or IgG synthesis, β agonists, tryptase inhibitors, aspirin, cyclooxygenase (COX) inhibitors, methotrexate, anti-TNF drugs, rituximab, inhibitors of PD4, p38 inhibitors, PDE4 inhibitors, and antihistamines, to name a few. The 2,4-pyrimidinediamine compounds can be administered by themselves in the form of prodrugs or as pharmaceutical compositions, which include an active compound.
[000260] In another embodiment, this invention provides a method of treating a Type IV hypersensitivity reaction, which includes administering to a patient an amount of a compound effective to treat or prevent the hypersensitivity reaction, where the compound is in accordance. with formula I as described herein. In one embodiment, the method is practiced prophylactically. In some embodiments, the compound is administered prior to exposure to an allergen.
[000261] In another embodiment, this invention provides a method of inhibiting a signal transduction cascade in which JAK3 kinase plays a role, which includes contacting a cell expressing a receptor involved in such a signaling cascade with a compound, where the compound is in accordance with formula I as described herein.
[000262] In another embodiment, this invention provides a method of treating a disease mediated by JAK kinase, which includes administering to a patient an amount compound effective to treat or prevent the disease mediated by JAK kinase, where the compound is of according to formula I as described herein.
[000263] In another embodiment, this invention provides a method of treating a JAK kinase mediated disease, wherein the JAK mediated disease is HVGR or GVHR, which includes administering to a patient an amount of compound effective to treat or prevent JAK kinase mediated disease, wherein the compound conforms to formula I as described herein.
[000264] In another embodiment, ocular disorders are treated using an effective amount of a compound of formula I as described herein. In one aspect of the disclosed method for treating ocular disorders, administration of one or more of the presently disclosed 2,4-pyrimidinediamine compounds is effective in increasing tear production volume as compared to untreated tear production volume, improving the latter. so a symptom of dry eye syndrome. In one aspect, tear production volume is increased within five days, such as within less than four days, and in some instances within less than two days. In one embodiment, the tear production volume is increased by at least about 25% over the initial tear production within two days of initial treatment with a presently disclosed 2,4-pyrimidinediamine compound. In other embodiments, tear production is increased by at least about 30%, such as at least about 50% over initial tear production within less than two days. Increases in tear production on administration of the present compounds result, in some cases, in tear production volume comparable to normal tear production. Typically the disclosed compounds, when used to treat eye disorders topically, are administered at least once a day and typically at most twice a day.
[000265] As mentioned, another embodiment provides a method of treating a disease and/or disorder of the eye, which includes administering to a patient an amount of a compound effective to treat the disease and/or disorder of the eye wherein the compound is in accordance with formula I as described herein. Diseases and disorders of the eye include, but are not limited to, dry eye syndrome, uveitis, allergic conjunctivitis, glaucoma, and rosacea (of the eye). Dry eye syndrome (DES), otherwise known as keratoconjunctivitis sicca (KCS), keratitis sicca, dry eye syndrome, or xerophthalmia, is an eye disease caused by decreased tear production or increased tear film evaporation commonly found in human beings. humans and some animals. Uveitis or iridocyclitis refers to inflammation of the middle layer of the eye (the “uvea”) and in common usage it can refer to any inflammatory process that involves the interior of the eye. Allergic conjunctivitis is inflammation of the conjunctiva (the membrane that covers the white part of the eye) due to an allergy. Glaucoma refers to a group of diseases that affect the optic nerve and involve a loss of retinal ganglion cells in a characteristic pattern, ie, a type of optic neuropathy. Elevated ocular pressure is a significant risk factor for the development of glaucoma (above 22 mmHg or 2.9 kPa), and inflammatory processes, eg uveitis, can cause this elevation in intraocular pressure. Rosacea is a chronic inflammatory condition characterized by facial erythema, but it can affect the eyes. As mentioned, the compounds described herein can be used to treat inflammatory responses. While not wishing to be bound by theory, the compounds described herein are believed to be effective treatments for these ocular disorders due, at least in part, to their JAK inhibitory activity.
[000266] In one embodiment, for the treatment of diseases and/or disorders of the eye, the compounds described herein, or their pharmaceutically acceptable salt forms are administered in combination or adjunct with at least one of an antihistamine, an antibiotic, an anti-inflammatory, an antiviral, and a glaucoma medication. Examples of common antibiotics used in the eye are sulfacetamide, erythromycin, gentamicin, tobramycin, ciprofloxacin and ofloxacin. Corticosteroids (sometimes referred to as “steroids”) are similar to a natural substance produced by the adrenal gland and are very effective anti-inflammatory drugs for a wide variety of eye problems. Corticosteroids can be safely used in the eye, and do not carry most of the risks associated with oral steroids like prednisone. Corticosteroids used to treat the eye include, but are not limited to, prednisolone, fluorometholone, and dexamethasone. Nonsteroidal anti-inflammatory drugs for the eye include, but are not limited to, ibuprofen, diclofenac, ketorolac, and flurbiprofen. Common antihistamines include livostin, patanol, cromolin, alomide. There are also over-the-counter antihistamines for the eye, which are less potent but can be very helpful in milder cases, such as pheniramine. Common antiviral eye medications include, but are not limited to, triflurthymidine, adenine, arabinoside, and idoxuridine. Glaucoma medications all try to reduce the eye's intraocular pressure, the fluid pressure within the eye, to prevent damage to the optic nerve that results in loss of vision. These medications can lower the pressure by decreasing the amount of fluid produced in the eye, by increasing the amount of fluid that leaves the eye's natural drain, or by providing additional pathways for fluid to leave the eye. Often more than one glaucoma medication will be used simultaneously, as these effects can combine to lower blood pressure even more than possible with just one medication. Medications against common glaucoma include, but are not limited to, beta blockers such as timolol, metipranolol, carteolol, betaxolol and levobunolol; prostaglandin analogues such as latanoprost; cholinergic agonists such as pilocarpine and carbachol; alpha agonists such as bromonidine and iopidine; carbonic anhydrase inhibitors such as dorzolamide; and adenergic agonists such as epinephrine and dipivefrin.
[000267] The active compounds described herein typically inhibit the JAK/Stat pathway. The activity of a specified compound as an inhibitor of a JAK kinase can be assessed in vitro or in vivo. In some embodiments, the activity of a specified compound can be tested in a cellular assay. Suitable assays include assays that determine the inhibition of the phosphorylation activity or ATPase activity of a JAK kinase. Thus, a compound is said to inhibit a JAK kinase activity if it inhibits the phosphorylation or ATPase activity of a JAK kinase with an IC50 of about 20 µM or less.
[000268] "Cell proliferative disorder" refers to a disorder characterized by abnormal proliferation of cells. A proliferative disorder does not imply any limitation with respect to the rate of cell growth, but merely indicates the loss of normal controls that affect cell growth and division. Thus, in some embodiments, cells of a proliferative disorder can have the same rates of cell division as normal cells but do not respond to signals that limit such growth. Within the scope of “cell proliferative disorder” is neoplasm or tumor, which is an abnormal growth of tissue. Cancer refers to any of a number of malignant neoplasms characterized by the proliferation of cells that have the ability to invade surrounding tissue and/or metastasize to new sites of colonization.
[000269] "Hematopoietic neoplasm" refers to a cell proliferative disorder arising from cells of the hematopoietic lineage. In general, hematopoiesis is the physiological process by which undifferentiated cells or stem cells develop into various cells found in peripheral blood. In the early developmental phase, hematopoietic stem cells, typically found in the bone marrow, undergo a series of cell divisions to form multipotent progenitor cells that engage in two main developmental pathways: the lymphoid lineage and the myeloid lineage. The involved progenitor cells of the myeloid lineage differentiate into three major sub-branches that include the developmental pathways of erythroid, megakaryocyte, and granulocyte/monocyte. An additional pathway leads to the formation of dendritic cells, which are involved in antigen presentation. The erythroid lineage gives rise to red blood cells while the megakaryocytic lineage gives rise to blood platelets. The involved cells of the granulocyte/monocyte lineage divide into developmental granulocyte or monocyte tracts, the first tract leading to the formation of neutrophils, eosinophils, and basophils and the last tract giving rise to blood monocytes and macrophages.
[000270] The involved progenitor cells of the lymphoid lineage develop in the B-cell pathway, the T-cell pathway, or the non-T/B-cell pathway. Similar to the myeloid lineage, an additional lymphoid pathway appears to give rise to dendritic cells involved in antigen presentation. The B cell progenitor cell develops into a precursor B cell (pre-B), which differentiates into B cells responsible for producing immunoglobulins. Progenitor cells of the T cell lineage differentiate into precursor T cells (pre-T) which, based on the influence of certain cytokines, develop into cytotoxic or helper/suppressor T cells involved in cell-mediated immunity. The non-T/B cell pathway leads to the generation of natural killer (NK) cells. Hematopoietic cell neoplasms can involve cells from any stage of hematopoiesis, which include hematopoietic stem cells, multipotent progenitor cells, oligopotent involved progenitor cells, precursor cells, and mature differentiated cells. The categories of hematopoietic neoplasms can generally follow the descriptions and diagnostic criteria used by those of skill in the art (see, for example, International Classification of Disease and Related Health Problems (ICD 10), World Health Organization (2003)). Hematopoietic neoplasms can also be characterized based on molecular characteristics, such as cell surface markers and gene expression profiles, the cell phenotype exhibited by aberrant cells, and/or chromosomal aberrations (eg, deletions, translocations, insertions, etc.) features of certain hematopoietic neoplasms, such as the Philadelphia chromosome found in chronic myelogenous leukemia. Other classifications include National Cancer Institute Working Formulation (Cancer, 1982, 49: 2112-2135) and Revised European-American Lymphoma Classification (REAL).
[000271] "Lymphoid neoplasm" refers to a proliferative disorder involving cells of the hematopoiesis lymphoid lineage. Lymphoid neoplasms may arise from hematopoietic stem cells as well as progenitor cells involved with lymphoid, precursor cells, and terminally differentiated cells. These neoplasms can be subdivided based on the phenotypic attributes of the aberrant cells or the differentiated state from which the abnormal cells arise. Subdivisions include, but are not limited to, B cell neoplasms, T cell neoplasms, NK cell neoplasms, and Hodgkin's lymphoma.
[000272] "Myeloid neoplasm" refers to the proliferative disorder of cells of the myeloid lineage hematopoiesis. Neoplasms may arise from hematopoietic stem cells, myeloid-involved progenitor cells, precursor cells, and terminally differentiated cells. Myeloid neoplasms can be subdivided based on the phenotypic attributes of the aberrant cells or the differentiated state from which the abnormal cells arise. Subdivisions include, but are not limited to, myeloproliferative disorders, myelodysplastic/myeloproliferative disorders, myelodysplastic syndromes, acute myeloid leukemia, and acute biphenotypic leukemia.
In general cell proliferative disorders treatable with the compounds disclosed herein refer to any disorder characterized by aberrant cell proliferation. These include various tumors and cancers, benign or malignant, metastatic or non-metastatic. Cancer-specific properties, such as tissue invasiveness or metastasis, can be targeted using the methods described herein. Cellular proliferative disorders include a variety of cancers, which include, but are not limited to, breast cancer, ovarian cancer, kidney cancer, gastrointestinal cancer, kidney cancer, bladder cancer, pancreatic cancer, squamous lung carcinoma, and adenocarcinoma. More specifically, related to particular tissues, organs or areas of the body, Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratomoa; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hanlartoma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), small intestine (adenocarcinoma, tumors carcinoids, Karposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor [nephroblastoma], lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratomoa, carcinoma embryonic, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor, chordoma, osteochronphoma (osteocartilaginous exostosis), chondroma chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma [ineblastaloma], glioblastoma, retinoglioma, schomandroma congenital tumors), spinal cord (neurofibroma, meningioma, glioma, sarcoma); Gynecological: uterus (endometrial carcinoma), cervix (cervical carcinoma, pretumor cervical dysplasia), ovaries (ovarian carcinoma [serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma], granulosa-thecal cell tumors, Sertoli-Leydig cell tumors , dysgerminoma, malignant teratomea), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonic rhabdomyosarcoma), fallopian tubes (carcinoma); Hematologic: blood (myeloid leukemia [acute and chronic], acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma [malignant lymphoma]; Skin: malignant melanoma, carcinoma of carcinoma basal cell, squamous cell carcinoma, Karposi's sarcoma, soft dysplastic nerve, lipoma, angioma, dermatofibroma, keloids, psori iasis; and Adrenal glands: neuroblastoma. The term "cancer cell" as provided herein includes a cell afflicted with any of the conditions identified above.
[000274] In some embodiments, the cell proliferative disorder treated is a hematopoietic neoplasm, which is the aberrant growth of cells of the hematopoietic system. Hematopoietic malignancies may originate from pluripotent stem cells, multipotent progenitor cells, involved oligopotent progenitor cells, precursor cells, and terminally differentiated involved cells in hematopoiesis. It is believed that some hematologic malignancies arise from hematopoietic stem cells, which have the capacity for self-renewal. For example, cells capable of developing specific subtypes of acute myeloid leukemia (AML) in transplantation demonstrate cell surface markers of hematopoietic stem cells, implying hematopoietic stem cells as the source of leukemic cells. Blast cells that do not have a hematopoietic stem cell marker characteristic appear to be unable to establish tumors on transplant (Blaire et al., 1997, Blood 89: 3104-3112). The stem cell origin of certain hematologic malignancies is also supported by the observation that specific chromosomal abnormalities associated with particular types of leukemia can be found in normal cells of the hematopoietic lineage as well as leukemic blast cells. For example, the reciprocal t(9q34;22q11) translocation associated with approximately 95% of chronic myelogenous leukemias appears to be present in cells of the myeloid, erythroid, and lymphoid lineage, suggesting that chromosomal aberration originates in hematopoietic stem cells. A subset of cells in certain types of CML demonstrate the hematopoietic stem cell marker cell phenotype.
[000275] Although hematopoietic neoplasms often originate from stem cells, involved progenitor cells or more terminally differentiated cells of a developmental lineage may also be the source of some leukemias. For example, the forced expression of the Bcr/Abl fusion protein (associated with chronic myelogenous leukemia) in common myeloid progenitor cells or granulocyte/macrophage progenitors produces a leukemic-equivalent condition. Furthermore, some chromosomal aberrations associated with leukemia subtypes are not found in the cell population with a hematopoietic stem cell marker phenotype, but are found in a cell population that demonstrates markers of one or more differentiated states of the hematopoietic pathway (Turhan et al. al., 1995, Blood 85: 2154-2161). Thus, although the involved progenitor cells and other differentiated cells may have only limited potential for cell division, leukemic cells may have acquired the ability to grow unregulated, in some cases mimicking the self-renewing characteristics of hematopoietic stem cells (Pass et al., Proc. Natl. Acad. Sci. USA, 2003, 100: 11842-9).
In some embodiments, the treated hematopoietic neoplasm is a lymphoid neoplasm, where the abnormal cells are derived from and/or demonstrate the characteristic phenotype of cells of the lymphoid lineage. Lymphoid neoplasms can be subdivided into B cell neoplasms, T and NK cell neoplasms, and Hodgkin's lymphoma. B cell neoplasms can be further subdivided into precursor B cell neoplasm and mature/peripheral B cell neoplasm. Exemplary B cell neoplasms are precursor B lymphoblastic leukemia/lymphoma (precursor B cell acute lymphoblastic leukemia) whereas exemplary mature/peripheral B cell neoplasms are B cell chronic lymphocytic leukemia/small lymphocytic lymphoma, B cell prolymphocytic leukemia , lymphoplasmacytic lymphoma, splenic marginal zone B cell lymphoma, hairy cell leukemia, plasma cell myeloma/plasmacytoma, MALT-type extranodal marginal zone B cell lymphoma, nodal marginal zone B cell lymphoma, follicular lymphoma, lymphoma of mantle cell, diffuse large B cell lymphoma, mediastinal large B cell lymphoma, primary effusion lymphoma, and Burkitt's lymphoma/Burkitt's cell leukemia. The presently disclosed compounds are particularly useful in the treatment of T cell and Nk cell neoplasms, which are further subdivided into precursor T cell neoplasm and mature (peripheral) T cell neoplasms. An exemplary precursor T cell neoplasm is precursor T lymphoblastic lymphoma/leukemia (precursor T cell acute lymphoblastic leukemia) while exemplary mature (peripheral) T cell neoplasms are T cell prolymphocytic leukemia, T cell granular lymphocytic leukemia, T cell leukemia aggressive NK cell, adult T cell lymphoma/leukemia (HTLV-1), extranodal NK/T cell lymphoma, nasal type T cell lymphoma, enteropathy type, hepatosplenic gamma-delta T cell lymphoma, T cell-equivalent lymphoma subcutaneous panniculitis, Mycosis fungoid/Sezary's syndrome, anaplastic large cell lymphoma, T cell/null, primary skin type, peripheral T cell lymphoma, not otherwise characterized, angioimmunoblastic T cell lymphoma, anaplastic large cell lymphoma, cell T/null, primary systemic type. The third limb of lymphoid neoplasms is Hodgkin's lymphoma, also referred to as Hodgkin's disease. Exemplary diagnoses of this class that can be treated with the compounds include, but are not limited to, nodular lymphocyte-predominant Hodgkin's lymphoma, and various classical forms of Hodgkin's disease, exemplary members of which are nodular sclerosis Hodgkin's lymphoma (grades 1 and 2 ), classical lymphocyte-rich Hodgkin's lymphoma, mixed cellularity Hodgkin's lymphoma, and lymphocyte-depleting Hodgkin's lymphoma. In various embodiments, any of the lymphoid neoplasms that are associated with aberrant JAK activity can be treated with the JAK inhibitor compounds.
[000277] In some embodiments, the treated hematopoietic neoplasm is a myeloid neoplasm. This group includes a large class of cell proliferative disorders involving or demonstrating the characteristic phenotype of myeloid lineage cells. Myeloid neoplasms can be subdivided into myeloproliferative diseases, myelodysplastic/myeloproliferative diseases, myelodysplastic syndromes, and acute myeloid leukemias. Exemplary myeloproliferative diseases are chronic myelogenous leukemia (eg, Philadelphia chromosome positive (t(9;22)(qq34;q11)), chronic neutrophilic leukemia, chronic eosinophilic leukemia/hypereosinophilic syndrome, chronic idiopathic myelofibrosis, polycythemia vera, and essential thrombocythemia Exemplary myelodysplastic/myeloproliferative diseases are chronic myelomonocytic leukemia, atypical chronic myelogenous leukemia, and juvenile myelomonocytic leukemia. refractory (myelodysplastic syndromes) with excess vlastics, 5q syndrome, and myelodysplastic syndromes In various embodiments, any of the myeloid neoplasms that are associated with aberrant JAK activity can be treated with the JAK inhibitor compounds.
[000278] In some embodiments, JAK inhibitor compounds can be used to treat acute myeloid leukemias (AML), which represent a large class of myeloid neoplasms having their own subdivision of disorders. These subdivisions include, but are not limited to, AMLs with recurrent cytogenetic translocations, AMLs with multilineage dysplasia, and other AMLs not otherwise categorized. Exemplary AMLs with recurrent cytogenetic translocations include, among others, AML with t(8;21)(q22;q22), AML1(CBF-alpha)/ETO, acute promyelocytic leukemia (AML with t(15;17)(q22;q11) -12) and variants, PML/RAR-alpha), AML with abnormal bone marrow eosinophils (inv(16)(p13q22) or t(16;16)(p13;q11), CBFb/MYH11X), and AML with abnormalities 11q23 (MLL). Exemplary AML with multilineage dysplasia are those that are associated with or without anterior myelodysplastic syndrome. Other acute myeloid leukemias not classified into any definable group include minimally differentiated AML, non-matured AML, maturing AML, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroid leukemia, acute megakaryocytic leukemia, acute basophilic leukemia, and panmyelosis.
[000279] One means of testing for such inhibition is to detect the effect of 2,4-pyrimidinediamine compounds on up-regulation of downstream gene products. In the Ramos/IL4 assay, B cells are stimulated with the cytokine Interleukin-4 (IL-4) leading to activation of the JAK/Stat pathway through phosphorylation of JAK family kinases, JAK1 and JAK3, which in turn phosphorylate and activate the transcription factor Stat-6. One of the genes upregulated by activated Stat-6 is the low-affinity IgE receptor, CD23. To study the effect of inhibitors (for example, the 2,4-substituted pyrimidinediamine compounds described herein) on JAK1 and JAK3 kinases, human Ramos B cells are stimulated with human IL-4. 20 to 24 hours after stimulation, cells are stained for CD23 upregulation and analyzed using FACS. A reduction in the amount of CD23 present compared to control conditions indicates that the test compound actively inhibits the JAK kinase pathway. An exemplary assay of this type is described in more detail in Example 2.
[000280] The activity of the compounds described herein can be further characterized by testing the effect of the 2,4-pyrimidinediamine compounds described herein on the proliferative responses of primary human T cells. In this assay, primary human T cells derived from peripheral blood and preactivated through stimulation of the T cell receptor and CD28, proliferate in culture in response to the cytokine Interleukin-2 (IL-2). This proliferative response is dependent on the activation of JAK1 and JAK3 tyrosine kinases, which phosphorylate and activate the transcription factor Stat-5. Primary human T cells are incubated with the 2,4-pyrimidinediamine compounds in the presence of IL-2 for 72 hours, and at the assay endpoint, intracellular ATP concentrations are measured to assess cell viability. A reduction in cell proliferation compared to control conditions is indicative of inhibition of the JAK kinase pathway.
[000281] The activity of the compounds described herein can be further characterized by testing the effect of the 2,4-pyrimidinediamine compounds described herein on A549 lung epithelial cells and U937 cells. A549 lung epithelial cells and U937 cells upregulate ICAM-1 (CD54) surface expression in response to a variety of different stimuli. Therefore, using ICAM-1 expression as readout, the effects of the test compound on different signaling pathways can be evaluated in the same cell type. Stimulation with IL-1β through IL-1β receptor activities activates the TRAF6/NFKB pathway resulting in upregulation of ICAM-1. IFNY induces ICAM-1 upregulation through activation of the JAK1/JAK2 pathway. ICAM-1 upregulation can be quantified by flow cytometry through a compound dose curve and EC50 values are calculated.
[000282] Active compounds as described herein generally inhibit the JAK kinase pathway with an IC50 in the range of about 1 mM or less, as measured in the assays described herein. Of course, a person of ordinary skill in the art would assess that compounds that exhibit lower IC50s, (in the order, for example, of 100 μM, 75 μM, 50 μM, 40 μM, 30 μM, 20 μM, 15 μM, 10 μM, 5 µM, 1 µM, 500 nM, 100 nM, 10 nM, 1 nM, or even lower) may be particularly useful in therapeutic applications. In cases where activity specific to a particular cell type is desired, the compound can be assayed for activity with the desired cell type and counter-screened for a lack of activity against other cell types. The desired degree of “inactivity” in such counter screens, or the desired ratio of activity vs. inactivity, can vary for different situations and can be selected by the user.
[000283] Active 2,4-pyrimidinediamine compounds also typically inhibit IL-4 stimulated expression of CD23 in B cells with an IC50 in the range of about 20 µM or less, typically in the range of about 10 µM, 1 µM, 500 nM, 100 nM, 10 nM, 1 nM, or even lower. A suitable assay that can be used is the assay described in Example 2, "Assay for IL-4 Stimulated Ramos B Cell Line." In certain embodiments, active 2,4-pyrimidinediamine compounds have an IC50 of less than or equal to 5 µM, greater than 5 µM but less than 20 µM, greater than 20 µM, or greater than 20 µM but less than 50 µM in the assay described in Example 2.
[000284] Additionally, active 2,4-pyrimidinediamine compounds typically inhibit a primary human T cell activity with an IC50 in the range of about 20 µM or less, typically in the range of about 10 µM, 1 µM, 500 nM, 100 nM, 10 nM, 1 nM, or even lower. The IC50 against human primary T cells can be determined in a standard in vitro assay with isolated human primary T cells. A suitable assay that can be used is the assay described above, "IL-2 Stimulated Primary Human T Cell Proliferation Assay." In some embodiments, active 2,4-pyrimidinediamine compounds have an IC50 of less than or equal to 5 µM, greater than 5 µM but less than 20 µM, greater than 20 µM, or greater than 20 µM but less than 50 µM in the assay described above.
[000285] Active 2,4-pyrimidinediamine compounds also typically inhibit ICAM1 (CD54) expression induced by exposure to IFNY in U937 or A549 cells with an IC50 in the range of about 20 µM or less, typically in the range of about of 10 μM, 1 μM, 500 nM, 100 nM, 10 nM, 1 nM, or even lower. The IC50 against ICAM (CD54) expression in IFNY-stimulated cells can be determined in a functional cell assay with an isolated A549 or U937 cell line. Active 2,4-pyrimidinediamine compounds typically have an IC50 of less than or equal to 20 μM, greater than 20 μM, or greater than 20 μM but less than 50 μM in the assay. Usefulness of compounds as research tools
[000286] A person of skill in the art would understand that certain crystallized protein-ligand complexes, in particular JAK-ligand complexes, and their corresponding X-ray structure coordinates can be used to reveal new structural information useful for understanding the biological activity of kinases as described herein. Also, the key structural features of the aforementioned proteins, particularly the shape of the ligand binding site, are useful in methods for designing or identifying selective kinase modulators and in resolving the structures of other proteins with similar characteristics. Such protein-ligand complexes, having compounds described herein as their ligand components, are an aspect of the invention.
[000287] Also, a person of skill in the art would appreciate that such suitable X-ray quality crystals can be used as part of a method of identifying a candidate agent capable of binding and modulating kinase activity. Such methods can be characterized by the following aspects: a) inputting into a suitable computer program information that defines a ligand binding domain of a kinase in a conformation (eg as defined by the X-ray structural coordinates obtained from crystals X-ray quality parameters as described above) where the computer program creates a model of the three-dimensional structures of the ligand binding domain, b) input a model of the three-dimensional structure of a candidate agent into the computer program, c) overlay the model of the candidate agent on the ligand binding domain model, and d) assessing whether the candidate agent model spatially fits into the ligand binding domain. Aspects a-d are not necessarily performed in the order mentioned above. Such methods can also entail: carrying out rational drug planning with the three-dimensional structure model, and selecting a potential candidate agent in conjunction with computer modeling.
[000288] Additionally, a person skilled in the art would appreciate that such methods may further entail: the use of a candidate agent, thus determined to spatially fit within the ligand binding domain, in a biological activity assay for kinase modulation, and determining whether said candidate agent modulates kinase activity in the assay. Such methods may also include administering the candidate agent, determined to modulate kinase activity, to a mammal suffering from a condition treatable by modulating the kinase, such as that described above.
[000289] Also, a person skilled in the art would appreciate that the compounds described herein can be used in a method of evaluating the ability of a test agent to associate with a molecule or molecular complex that includes a ligand binding domain of a kinase . Such a method can be characterized by the following aspects: a) creating a computer model of a kinase binding pocket using structural coordinates obtained from suitable x-ray quality crystals of the kinase, b) using computational algorithms to perform an adjustment operation between the test agent and the binding pouch computer model, and c) analyzing the results of the fitting operation to quantify the association between the test agent and the binding pouch computer model. Utility of compounds as screening agents
[000290] To use the compounds described herein in a method of screening for candidate agents that bind to, for example, a JAK protein, the protein is attached to a support, and a compound described herein is added to the assay. Alternatively, the compound described herein is attached to the support, for example via a linker that does not prohibitively affect biological activity, and the protein is added. Classes of candidate agents among which new binding agents can be searched include specific antibodies, unnatural binding agents identified in chemical library screens, peptide analogs, etc. Of particular interest are screening assays for candidate agents that have low toxicity to human cells. A wide variety of assays can be used for this purpose, which include labeled in vitro protein-protein binding assays, electrophoretic mobility shift assays, immunoassays for protein binding, functional assays (phosphorylation assays, etc.) and others .
[000291] The determination of candidate agent binding, for example, to a JAK protein can be done in several ways. In one example, the candidate agent (the compound described herein) is labeled, for example, with a fluorescent or radioactive moiety and binding directly determined. For example, this can be done by attaching all or a portion of the JAK protein to a solid support, adding a labeled agent (for example a compound described herein in which at least one atom has been replaced by a detectable isotope), removing the reagent. in excess by washing, and determining whether the amount of the label is that present on the solid support. Various blocking and washing steps can be used as is known in the art. "Labeled" means that the compound is directly or indirectly labeled with something that provides a detectable signal, for example, radioisotope, fluorescent label, enzyme, antibodies, particles such as magnetic particles, chemiluminescent labels, or specific binding molecules, etc. specific binding molecules include pairs such as biotin and streptavidin, digoxin and antidigoxin etc. For specific binding members, the complementary member would normally be labeled with a molecule that provides detection, in accordance with known procedures, as outlined above. The label can directly or indirectly provide a detectable signal.
[000292] In some embodiments, only one of the components is labeled. For example, a JAK protein can be labeled at tyrosine positions using125I, or with fluorophores. Alternatively, more than one component can be labeled with different labels; using 125I for proteins, for example, and a fluorophore for candidate agents.
[000293] The compounds described herein can also be used as competitors to screen for additional drug candidates. "Candidate bioactive agent" or "drug candidate" or grammatical equivalents as used herein describe any molecule, e.g., protein, oligopeptide, small organic molecule, polysaccharide, polynucleotide, etc., to be tested for bioactivity. They may be able to directly or indirectly alter the cell proliferation phenotype or the expression of a cell proliferation sequence, which includes both nucleic acid sequences and protein sequences. In other cases, alteration of cell proliferation protein binding and/or activity is screened for. In the case where protein binding or activity is screened, some embodiments exclude molecules already known to bind to that particular protein. Exemplary embodiments of assays described herein include candidate agents, which do not bind to the target protein in its native endogenous state, referred to herein as "exogenous" agents. In one example, exogenous agents further exclude antibodies to JAK proteins.
[000294] Candidate agents can span numerous chemical classes, although typically they are organic molecules having a molecular weight of more than about 100 daltons and less than about 2,500 daltons. Candidate agents include functional groups necessary for structural interaction with proteins, particularly hydrogen bonding and lipophilic bonding, and typically include at least one amine, carbonyl, hydroxyl, ether, or carboxyl group, for example at least two of the functional chemical groups. Candidate agents often include cyclic carbon or heterocyclyl structures and/or aromatic or polyaromatic structures substituted with one or more of the above functional groups. Candidate agents are also found among biomolecules that include peptides, saccharides, fatty acids, steroids, purines, pyrimidines, derivatives, structural analogs, or combinations thereof.
[000295] Candidate agents are obtained from a wide variety of sources including libraries of synthetic or natural compounds. For example, numerous means are available for the random and targeted synthesis of a wide variety of organic compounds and biomolecules, which include the expression of randomized oligonucleotides. Alternatively, libraries of natural compounds in the form of bacterial, fungal, plant and animal extracts are available or easily produced. Additionally, libraries of natural or synthetically produced compounds are easily modified through conventional chemical, physical and biochemical means. Known pharmacological agents can be subjected to targeted or random chemical modifications such as acylation, alkylation, esterification, amidification to produce structural analogs.
[000296] In one example, candidate agent binding is determined through the use of competitive binding assays. In this example, the competitor is a binding moiety known to bind to a JAK protein, such as an antibody, peptide, binding partner, ligand, etc. Under certain circumstances, there may be competitive binding as between the candidate agent and the binding portion, with the binding portion displacing the candidate agent.
[000297] In some embodiments, the candidate agent is labeled. The candidate agent, or the competitor, or both, are first added for example to a JAK protein for a time sufficient to allow binding, if present. Incubations can be performed at any temperature that facilitates optimal activity, typically between 4°C and 40°C. Incubation periods are selected for optimal activity, but can also be optimized to facilitate rapid high-performance screening. Excess reagent is usually removed or washed away. The second component is then added, and the presence or absence of the labeled component is tracked to indicate binding.
[000298] In one example, the competitor is added first, followed by the candidate agent. Competitor displacement is an indication that the candidate agent is bound to a JAK protein and thus is capable of binding to, and potentially modulating, the activity of the JAK protein. In this embodiment, the component can be labeled. So, for example, if the competitor is labeled, the presence of a label in the wash solution indicates displacement by the agent. Alternatively, if the candidate agent is labeled, the presence of the label on the support indicates displacement.
[000299] In an alternative embodiment, the candidate agent is added first, with incubation and washing, followed by the competitor. The absence of binding by the competitor may indicate that the candidate agent is bound to a JAK protein with a higher affinity. Thus, if the candidate agent is labeled, the presence of the label on the support, linked with a lack of competitor binding, may indicate that the candidate agent is capable of binding to the JAK protein.
[000300] It may be of value to identify the binding site of a JAK protein. This can be done in a variety of ways. In one embodiment, once the JAK protein has been identified as binding to the candidate agent, the JAK protein is fragmented or modified and the assays repeated to identify components required for binding.
[000301] Modulation is tested by screening for candidate agents capable of modulating the activity of a JAK protein which includes the steps of combining a candidate agent with the JAK protein, as above, and determining an alteration in the biological activity of the JAK protein. Thus, in this embodiment, the candidate agent must either bind (although this may not be necessary), or alter its biological or biochemical activity as defined herein. Methods include both in vitro screening methods and in vivo screening of cells for changes in cell viability, morphology, and others.
[000302] Alternatively, differential screening can be used to identify drug candidates that bind to a native JAK protein but cannot bind to a modified JAK protein.
[000303] A variety of other reagents can be included in screening assays. These reagents include as salts, neutral proteins, eg albumin, detergents, etc. which can be used to facilitate optimal protein-protein binding and/or reduce non-specific or background interactions. Also reagents that otherwise improve the efficiency of the assay, such as protease inhibitors, nuclease inhibitors, antimicrobial agents, etc., can be used. The component mix can be added in any order that provides the necessary bond. Administration Methods
[000304] The 2,4-pyrimidinediamine compound(s) or prodrug(s) described herein, or compositions thereof, in general will be used in an effective amount to obtain the desired result, for example, in an amount effective to treat or prevent the particular condition being treated. The compound(s) may be administered therapeutically to obtain therapeutic benefit or prophylactically to obtain prophylactic benefit. By therapeutic benefit is intended the eradication or amelioration of the underlying disorder being treated and/or eradication or amelioration of one or more of the symptoms associated with the underlying disorder such that the patient reports an improvement in the sensation or condition, notwithstanding the patient may be still afflicted with the underlying disorder. For example, administering a compound to a patient suffering from an allergy provides therapeutic benefit not only when the underlying allergic response is eradicated or ameliorated, but also when the patient reports a decrease in the severity or duration of symptoms associated with the allergy to following exposure to the allergen. As another example, therapeutic benefit in the context of asthma includes an improvement in breathing following the onset of an asthmatic attack or a reduction in the frequency or severity of asthmatic episodes. As another specific example, the therapeutic benefit in the context of transplant rejection includes the ability to alleviate an episode of acute rejection, such as, for example, HVGR or GVHR, or the ability to prolong the time period between the onset of episodes. of acute rejection and/or onset of chronic rejection. Therapeutic benefit also includes halting or slowing disease progression, regardless of whether improvement is achieved.
[000305] The amount of compound administered will depend on a variety of factors, including, for example, the particular condition being treated, the mode of administration, the severity of the condition being treated, the age and weight of the patient, the bioavailability of the particular active compound. Determining an effective dosage is well within the capabilities of those skilled in the art.
[000306] As known to those of skill in the art, the preferred dosage of 2,4-pyrimidinediamine compounds will also depend on the age, weight, general health, and severity of the condition of the individual being treated. The dosage may also need to be adapted to the individual's sex and/or the individual's lung capacity, where administered by inhalation. The dosage can also be adapted for individuals who suffer from more than one condition or those individuals who have additional conditions that affect lung capacity and the ability to breathe normally, for example, emphysema, bronchitis, pneumonia, and respiratory infections. The dosage, and frequency of administration of the compounds or prodrugs thereof, will also depend on whether the compounds are formulated for the treatment of acute episodes of a condition or for the prophylactic treatment of a disorder. For example, acute episodes of allergic conditions, which include allergy-related asthma, transplant rejection, etc. A skilled technician will be able to determine the optimal dose for a particular individual.
[000307] For prophylactic administration, the compound can be administered to a patient at risk for developing one of the conditions described above. For example, if it is unknown whether a patient is allergic to a particular drug, the compound can be administered prior to administration of the drug to prevent or ameliorate an allergic response to the drug. Alternatively, prophylactic administration can be applied to prevent the onset of symptoms in a patient diagnosed with the underlying disorder. For example, a compound can be administered to an allergy sufferer before expected exposure to the allergen. The compounds can also be administered prophylactically to healthy individuals who are repeatedly exposed to agents known to one of the ailments described above to prevent the onset of the disorder. For example, a compound can be administered to a healthy individual who is repeatedly exposed to an allergen known to induce allergies, such as latex, in an effort to prevent the individual from developing an allergy. Alternatively, a compound can be administered to a patient suffering from asthma before participating in activities that trigger asthma attacks to lessen the severity of, or completely prevent, an asthmatic episode.
[000308] In the context of transplant rejection, the compound can be administered while the patient is not having an acute rejection reaction to prevent the onset of rejection and/or before the appearance of clinical indications of chronic rejection. The compound can be systemically administered to the patient as well as administered to the tissue or organ prior to transplanting the tissue or organ into the patient.
[000309] The amount of compound administered will depend on a variety of factors, including, for example, the particular indication being treated, the mode of administration, whether the benefit desired is prophylactic or therapeutic, the severity of the indication being treated and the age and weight of the patient, and the bioavailability of the particular active compound. Determining an effective dosage is well within the capabilities of those skilled in the art.
[000310] Effective dosages can be estimated initially from in vitro assays. For example, an initial dosage for use in animals can be formulated to achieve a circulating blood or serum concentration of active compound, that is, at or above an IC50 of the particular compound as measured in an in vitro assay. Calculating dosages to obtain such circulating blood or serum concentrations taking into account the bioavailability of the particular compound is well within the capabilities of skilled practitioners. For guidance, the reader is referred to Fingl & Woodbury, “General Principles,” In: Goodman and Gilman's The Pharmaceutical Basis of Therapeutics, Chapter 1, pp. 1-46, last edition, Pergamagon Press, and references cited therein.
[000311] Initial dosages can also be estimated from in vivo data such as animal models. Animal models useful for testing the effectiveness of compounds to treat or prevent the various diseases described above are well known in the art. Suitable animal models of hypersensitivity or allergic reactions are described in Foster, (1995) Allergy 50(21Suppl): 6-9, debate 34-38 and Tumas et al., (2001), J. Allergy Clin. Immunol. 107(6): 1025-1033. Suitable animal models of allergic rhinitis are described in Szelenyi et al., (2000), Arzneimittelforschung 50(11): 1037-42; Kawaguchi et al., (1994), Clin. Exp. Allergy 24(3): 238-244 and Sugimoto et al., (2000), Immunopharmacology 48(1): 1 - 7. Suitable animal models of allergic conjunctivitis are described in Carreras et al., (1993) , Br.J. Ophthalmol. 77(8): 509-514; Saiga et al., (1992), Ophthalmic Res. 24(1): 45-50; and Kunert et al., (2001), Invest. Ophthalmol. Vis. Sci. 42(11): 2483-2489. Suitable animal models of systemic mastocytosis are described in O'Keefe et al., (1987), J. Vet. Intern. Med. 1(2): 75-80 and Bean-Knudsen et al., (1989), Vet. Pathol. 26(1): 90-92. Suitable animal models of hyper IgE syndrome are described in Claman et al., (1990), Clin. Immunol. Immunopathol. 56(1): 46-53. Suitable animal models of B-cell lymphoma are described in Hough et al., (1998), Proc. Natl. Academic Sci. USA 95: 13853-13858 and Hakim et al., (1996), J. Immunol. 157(12): 5503-5511. Suitable animal models of atopic disorders such as atopic dermatitis, atopic eczema, and atopic asthma are described in Chan et al., (2001), J. Invest. Dermatol. 117(4):977-983 and Suto et al., (1999), Int. Arch. Allergy Immunol. 120 (Suppl 1): 70 - 75. Suitable animal models of transplant rejection, such as HVGR models, are described in O'Shea et al., (2004), Nature Reviews Drug Discovery 3: 555564; Cetkovic-Curlje & Tibbles, (2004), Current Pharmaceutical Design 10: 1767-1784; and Chengelian et al., (2003), Science 302:875-878. Ordinarily skilled technicians may routinely adapt such information to determine dosages suitable for human administration.
[000312] Dosage amounts will typically be in the range of about 0.0001 or 0.001 or 0.01 mg/kg/day to about 100 mg/kg/day, but may be higher or lower, depending, between other factors, the activity of the compound, its bioavailability, the mode of administration, and various factors discussed above. The dosage amount and interval can be adjusted individually to provide plasma levels of the compound(s) which is/are sufficient to maintain the therapeutic or prophylactic effect. For example, the compounds can be administered once a week, several times a week (e.g., every second day), once a day, or multiple times a day, depending on, among other things, the mode of administration, the specific indication that is treated, and the judgment of the prescribing physician. In cases of local administration or selective uptake, such as local topical administration, the effective local concentration of active compound(s) may not be related to the plasma concentration. A person of ordinary skill in the art would be able to optimize effective local dosages without undue experimentation.
[000313] Preferably, the compound(s) will provide therapeutic or prophylactic benefit without causing substantial toxicity. The toxicity of the compound(s) can be determined using standard pharmaceutical procedures. The dose ratio between toxic and therapeutic (or prophylactic) effect is the therapeutic index. Compound(s) which exhibit(s) high therapeutic indices are preferred.
[000314] The foregoing disclosure regarding the dosage requirements for 2,4-substituted pyrimidinediamine compounds is pertinent to the dosages required for prodrugs, with the finding, evident to skilled technicians, that the prodrug amount( s) administered will also depend on a variety of factors, which include, for example, the bioavailability of the particular prodrug(s) and the rate of conversion and efficiency into an active drug compound under the chosen administration. Determination of an effective dosage of prodrug(s) for a particular use and mode of administration is well within the ability of those skilled in the art.
[000315] Effective dosages can be estimated initially from assays of in vitro activity and metabolism. For example, an initial dosage of prodrug for use in animals can be formulated to obtain a circulating blood or serum concentration of the active metabolite compound i.e. at or above an IC50 of the particular compound as measured in an in vitro assay, such as the in vitro CHMC or BMMC and other in vitro assays described in US Serial Application No. 10/355,543 filed January 31, 2003 (US2004/0029902A1), International Application Serial No. PCT/US03/03022 filed January 31, 2003 (WO 03/063794), US Serial Application No. 10/631,029 filed July 29, 2003, International Serial Application No. PCT/US03/24087 (WO2004/014382), US Serial Application No. 10/903,263 filed July 30 2004, and International Serial Application No. PCT/US2004/24716 (WO005/016893). Calculating dosages to obtain such circulating blood or serum concentrations, taking into account the bioavailability of the particular prodrug via the desired route of administration, is well within the capabilities of a person of ordinary skill in the art. For guidance, the reader is referred to Fingl & Woodbury, “General Principles,” In: Goodman and Gilman's The Pharmaceutical Basis of Therapeutics, Chapter 1, pp. 1-46, last edition, Pagamonon Press, and references cited therein.
[000316] Also provided are kits for the administration of 2,4-pyrimidinediamine, prodrug thereof, or pharmaceutical formulations which include the compound which may include a dosage amount of at least one 2,4-pyrimidinediamine or a composition which includes at least one minus one 2,4-pyrimidinediamine, as disclosed herein. Kits may also include appropriate packaging and/or instructions for using the compound. Kits may also include a means for delivering the at least one 2,4-pyrimidinediamine or compositions that include at least one 2,4-pyrimidinediamine, such as an inhaler, spray dispenser (e.g., nasal spray), syringe for injection, or pressurized packaging for capsules, tablets, suppositories, or other device as described herein. A kit can also provide the compound and reagents to prepare a composition for administration. The composition may be in a dry or lyophilized form or in a solution, particularly a sterile solution. When the composition is in a dry form, the reagent can include a pharmaceutically acceptable diluent to prepare a liquid formulation. The kit can contain a device for administering or dispensing the compositions, which include, but are not limited to, a syringe, pipette, transdermal patch, or inhalant.
[000317] Kits may include other therapeutic compounds for use in conjunction with the compounds described herein. In one embodiment, the therapeutic agents are immunosuppressive or anti-allergen compounds. These compounds can be provided in a separate form or mixed with the compounds of the present invention.
[000318] The kits will include appropriate instructions for the preparation and administration of the composition, the side effects of the compositions, and any other relevant information. Instructions may be in any suitable format, including, but not limited to, printed matter, videotape, computer-readable disc, or optical disc.
[000319] One embodiment is a kit that includes a compound of formula I, or a prodrug thereof, packaging, and instructions for use.
[000320] In another embodiment, this invention provides a kit that includes a pharmaceutical formulation that includes a compound of formula I or a prodrug thereof and at least one pharmaceutically acceptable excipient, diluent, preservative, stabilizer, or mixture thereof , packaging, and instructions for use.
[000321] Another embodiment a kit for treating an individual suffering from or susceptible to the conditions described herein are provided, which includes a container that includes a dosage amount of a 2,4-pyrimidinediamine or composition as disclosed herein. , and instructions for use. The container can be any of those known in the art and suitable for the storage and release of oral, intravenous, topical, rectal, urethral, or inhaled formulations.
Kits may also be provided containing sufficient dosages of the 2,4-pyrimidinediamine or composition to provide effective treatment to an individual for an extended period, such as 1 week, 2 weeks, 3, weeks, 4 weeks, 6 weeks, or 8 weeks or more.
[000323] It will be appreciated by a person of skill in the art that the embodiments summarized above may be used together in any suitable combination to generate additional embodiments not expressly stated above, and that such embodiments are considered to be part of the present invention. Manufacturing Methods
[000324] The 2,4-pyrimidinediamine compounds described herein can be synthesized via a variety of different synthetic routes using commercially available starting materials and/or starting materials prepared by conventional synthetic methods. Suitable exemplary methods that can be routinely adapted to synthesize the 2,4-pyrimidinediamine compounds and prodrugs described herein are found in U.S. Patent No. 5,958,935, the disclosure of which is incorporated herein by reference. Specific examples describing the synthesis of numerous 2,4-pyrimidinediamine compounds and prodrugs, as well as intermediates thereafter, are described in US Serial No. 10/355,543, filed January 31, 2003 (US2004/0029902A1), the contents of which are incorporated herein by reference. Suitable exemplary methods that can be routinely used and/or adapted to synthesize active 2,4-substituted pyrimidinediamine compounds can also be found in international application Serial No. PCT/US03/03022 filed January 31, 2003 (WO 03/063794 ), US Serial Application No. 10/631,029 filed July 29, 2003, International Serial Application No. PCT/US03/24087 (WO2004/014382), US Serial Application No. 10/903,263 filed July 30, 2004, and International Application Serial No. PCT/US2004/24716 (WO005/016893), the disclosures of which are incorporated herein by reference. All compounds described herein (which include prodrugs) can be prepared by routine adaptation of these methods.
[000325] Exemplary synthetic routes that can be used to synthesize the 2,4-pyrimidinediamine compounds described herein are depicted in Schemes (I) through (II), below. These methods can be routinely adapted to synthesize the 2,4-substituted pyrimidinediamine compounds described herein. After each reaction step, the product can be purified or can, depending on the chemistry, be used in the next step without purification. Exemplary methods for making the 2,4-substituted pyrimidinediamines described herein are also included in the examples below. Those of skill in the art will also be able to readily adapt these examples for the synthesis of additional 2,4-substituted pyrimidinediamines as described herein.
[000326] The compounds disclosed herein can be synthesized for example from substituted or unsubstituted uracils as illustrated in Scheme (I) below. In Scheme (I), ring A, R5, (R2)p, X, Y, Z1, and Z2 are as defined herein. According to Scheme (I), uracil A-1 is dialyzed at positions 2 and 4 using a standard halogenating agent such as POCl 3 (or other standard halogenating agent) under standard conditions to produce 2,4-dichloropyrimidine A -two. Depending on the R5 substituent, in pyrimidinediamine A-2, the chloride at the C4 position may be more reactive towards the nucleophiles than the chloride at the C2 position. This differential reactivity can be exploited to synthesize 2,4-pyrimidinediamines I, for example when R5 is F, by first reacting 2,4-dichloropyrimidine A-2 with an equivalent of amine A-3, producing 2-chloro 4N-substituted-4-pyrimidineamine A-4, followed by amine A-5 to produce a 2,4-pyrimidinediamine of formula A-6 (compounds of formula I, where each of R3 and R4 are H). Compounds of formula I, where either or both of R3 and R4 are not H, can be manufactured, for example, by alkylating the NH groups at C2 or C4 of pyrimidine-2,4-diamine. Scheme (I)

[000327] Typically, the C4 halide is more reactive towards the nucleophiles. However, as will be recognized by one of ordinary skill in the art, the identity of the R5 substituent can alter this reactivity. For example, when R5 is trifluoromethyl, a 50:50 mixture of 4N-substituted 4-pyrimidineamine A-4 and the corresponding 2N-substituted 2-pyrimidineamine is obtained (not shown). The regioselectivity of the reaction can also be controlled by adjusting the solvent and other synthetic conditions (such as temperature), as is well known in the art. Alternative synthetic methods allow for complete regioselectivity, for example, when manufacturing A-6 molecules. One such method is described in relation to Scheme (II) below.
[000328] The reactions depicted in Scheme (I) to make A-4 and A-6 (and A-6 in Scheme (II) below) can process more quickly when the reaction mixtures are heated via microwave. When heating in this manner, the following conditions can be used: heat up to 175°C in ethanol for 5 to 20 min. in a Smith Reactor (Personal Chemistry, Uppsala, Sweden) in a sealed tube (at 20 bar pressure).
[000329] Uracil A-1 starting materials can be purchased from commercial sources or prepared using standard techniques of organic chemistry. Commercially available uracils that can be used as starting materials in Scheme (I) include, By way of example and not limitation, uracil (Aldrich #13.078-8; CAS Registry 66-22-8); 5- bromouracil (Aldrich #85.247-3; CAS Registry 51-20-7; 5-fluorouracil (Aldrich #85.847-1; CAS Registry 51-21-8); 5-iodouracil (Aldrich #85,785-8; CAS Registry 696 -07-1); 5-nitrouracil (Aldrich #85.276-7; CAS Registry 611-08-5); 5-(trifluoromethyl)-uracil (Aldrich #22.327-1; CAS Registry 5420-6). additional substitutes are available from General Intermediates of Canada, Inc., Edmonton, CA and/or Interchim, Cedex, France, or can be prepared using standard techniques.A large number of textbook references disclosing suitable synthetic methods are provided below.
[000330] Amines A-3 and A-5 can be purchased from commercial sources or, alternatively, can be synthesized using standard techniques. For example, suitable amines can be synthesized from nitro precursors using standard chemistry. Specific exemplary reactions used to make anilines are given in the Examples section. See also Vogel, 1989, Practical Organic Chemistry, Addison Wesley Longman, Ltd. and John Wiley & Sons, Inc.
[000331] One of ordinary skill in the art would recognize that in some cases, the amines A-3 and A-5 and/or the X substituent on the A-1 uracil may include functional groups that require protection during synthesis. The exact identity of any protection group(s) used will depend on the identity of the functional group that is protected, and will be evident to those of skill in the art. Guidance on the selection of appropriate protecting groups, as well as synthetic strategies for their attachment and removal, can be found, for example, in Green & Wuts.
[000332] Thus, protecting group refers to a group of atoms that, when attached to a reactive functional group in a molecule, masks, reduces or prevents the reactivity of the functional group. Typically, a protecting group can be selectively removed as desired during the course of a synthesis. Examples of protecting groups can also be found in Harrison et al., Compendium of Synthetic Organic Methods, Vols. 1-8, 1971-1996, John Wiley & Sons, NY. Representative amino protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl ("CBZ"), tert-butoxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-trimethylsilyl- ethanesulfonyl ("TES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl ("FMOC"), nitro-veratryloxycarbonyl ("NVOC") and others. Representative hydroxyl protecting groups include, but are not limited to those where the hydroxyl group is acylated to form acetate and benzoate esters or alkylated to form benzyl and trityl ethers, as well as alkyl ethers, tetrahydropyranyl ethers, ethers. trialkylsilyl (eg TMS or TIPPS groups) and allyl ethers.
[000333] A large number of references disclosing useful methods for synthesizing pyrimidines in general, as well as starting materials described in the Schemes are known in the art. For specific guidance, the reader is referred to Brown, DJ, “The Pirimidines,” in The Chemistry of Heterocyclic Compounds, Volume 16 (Weissberger, A., Ed.), 1962, Interscience Publishers, (A Division of John Wiley & Sons ), New York (“Brown I”); Brown, DJ, "The Pyrimidines", in The Chemistry of Heterocyclic Compounds, Volume 16, Supplement I (Weissberger, A. and Tailor, EC, Ed.), 1970, Wiley-Interscience, (A Division of John Wiley & Sons) , New York (Brown II”); Brown, DJ, “The Pirimidines,” in The Chemistry of Heterocyclic Compounds, Volume 16, Supplement II (Weissberger, A. and Tailor, EC, Ed.), 1985, A Interscience Publication (John Wiley & Sons), New York ( “Brown III”); Brown, D.J., "The Pyrimidines" in The Chemistry of Heterocyclic Compounds, Volume 52 (Weissberger, A. and Tailor, E.C., Ed.), 1994, John Wiley & Sons, Inc., New York, pp. 1-1509 (Brown IV"); Kenner, G.W. and Todd, A., in Heterocyclic Compounds, Volume 6, (Elderfield, R.C., Ed.), 1957, John Wiley, New York, Chapter 7 (pyrimidines); Paquette, L.A., Principles of Modern Heterocyclic Chemistry, 1968, W.A. Benjamin, Inc., New York, pp. 1 - 401 (synthesis of uracil pp. 313, 315; synthesis of pyrimidinediamine pp. 313-316; synthesis of amino pyrimidinediamine pp. 315); Joule, J.A., Mills, K. and Smith, G.F., Heterocyclic Chemistry, 3rd Edition, 1995, Chapman and Hall, London, UK, pp. 1 - 516; Vorbrüggen, H. and Ruh-Pohlenz, C., Handbook of Nucleoside Synthesis, John Wiley & Sons, New York, 2001, pp. 1-631 (protection of pyrimidines by acylation pp. 90-91; silylation of pyrimidines pp. 91-93); Joule, J.A., Mills, K. and Smith, G.F., Heterocyclic Chemistry, 4th Edition, 2000, Blackwell Science, Ltd, Oxford, UK, pp. 1 - 589; and Comprehensive Organic Synthesis, Volumes 1-9 (Trost, B.M. and Fleming, I., Ed.), 1991, Pergamon Press, Oxford, UK.
[000334] A specific embodiment of Scheme (I) which uses 5-fluorouracil (Aldrich #32,937-1) as a starting material is illustrated in Scheme (Ia), below. In scheme (Ia), ring A, (R2)p, X, Y, Z1, and Z2 are as previously defined for Scheme (I) and formula I. Compound A-10, a 5-fluoro-2, 2N,4N-disubstituted 4-pyrimidinediamine, can be obtained by reacting 2,4-dichloro-5-fluoropyrimidine A-8 (commercially available or manufactured from A-7 as shown for example, starting with an uracil and dehydrohalogenation with for example POCl3) with optimally one equivalent of the amine A-3 to produce the 2-chloro-N4-substituted-5-fluoro-4-pyrimidineamine A-9 followed by reaction with one or more equivalents of the amine A-5 , typically between about 1.1 equivalents of A-5 and about 2 equivalents of A-5. The reaction of A-9 with A-5 can be carried out, for example, by traditional heating, by microwave irradiation and by Buchwald type bonding using metal catalyst, eg palladium bonding. The last of the metal-catalyzed bonding reactions is typically used when the nucleophilic partner, A-5, is insufficiently nucleophilic, and so reactions where bond insertions and reductive bonds work well are used to effect the transformation. Scheme (Ia)

[000335] Although many of the synthetic schemes discussed above do not illustrate the use of protecting groups, a person of ordinary skill in the art would recognize that in some cases certain substituents, such as, for example, R2 and/or other groups, may include functionality that requires protection. The exact identity of the protecting group used will depend, among other things, on the identity of the functional group that is protected and the reaction conditions used in the particular synthetic scheme, and will be evident to those of skill in the art. Guidance on the selection of protective groups, their attachment and removal suitable for a particular application can be found, for example, in Green & Wuts.
Schemes I and Ia above describe the synthesis of the 2,4-pyrimidinediamines described herein by the addition of two amine nucleophiles to a pyrimidine dielectrophil, for example a 2,4-dialopyrimidine. The compounds disclosed herein, including those according to Formula I as described herein and including compounds of Formulas IA, IB, IC, II, III and IV can be synthesized according to Schemes I and Ia. can also be synthesized according to Scheme II below describes a new synthesis of the compounds described herein starting from the A ring anilines, building the pyrimidine system into the aniline via the guanyl analogues, and then installing the 4N-ring system heteroaryl. First, amines A-5 are converted to the corresponding guanidine derivatives A-11. For example, amine A-5 is reacted with an appropriately protected electrophilic guanyl partner, for example N,N-bis-Boc-1-guanylpyrazole, for example, in the presence of a base, for example a tertiary amine base. The reaction is typically, but not necessarily, heated to between about 40°C and about 60°C for between about 24 h and about 48 h. After this step, any of the protecting groups are removed. Scheme (II)

[000337] For example, when N,N-bis-Boc-1-guanylpyrazole is used, an acid is added to the mixture (optionally volatiles are removed from the first reaction) to remove the Boc protecting groups and provide A-11. The guanidines A-11 are reacted with the appropriate 1,3-dielectrophils, for example β-aldehyde esters, A-12 (where Re is defined as in formula I), for example by heating the guanidine in an alcoholic solvent in the presence of base. In one example, sodium ethoxide in ethanol is used. The reaction is heated to between about 40°C and about 80°C for between about 24 h and about 48 h to give the corresponding 4-hydroxy-pyrimidinediamino-2-amine, A-13. A person of ordinary skill in the art would appreciate that other 1,3-dielectrophils can be used to make the A-13 pyrimidine intermediates, for example, 2-substituted cyanoacetaldehydes and α,β-unsaturated esters such as 2-(acetoxymethyl)acrylates of alkyl (Bailis-Hillman acetates, for example as reported in Bull. Korean Chemical Soc. 2007, Vol. 28, No. 12, 2505-2507, which is incorporated herein by reference for all purposes), and the like. The intermediates of A-13 can be purified in some cases or used "as such" in the next reaction, where they are converted to the corresponding 4-leaving group-pyrimidinediamino-2-amines, A-14. For example, intermediates A-13 can be reacted with, for example, POCl3 to provide the corresponding 4-chloro-pyrimidine-2-amine, i.e., amines A-14 where LG is chlorine. A person of ordinary skill in the art would appreciate what other starting groups would work, for example, the hydroxy group of intermediate A-13 can be converted to a mesylate and then used for the next step, for example, in place of a 4-halo intermediate . Compounds A-6 (compounds of formula I where R3 and R4 are H) are formed by reacting intermediate A-14 with amines A-3, for example, as described above in Schemes I and Ia in connection with the reaction with amines. intermediates A-2 and/or A-8, respectively. Compounds of formula I, where R3 and/or R4 are not H, are manufactured, for example, by the alkylation of compounds A-6 with the appropriate alkylating reagents, as would be appreciated by a person of ordinary skill in the art. Alternatively, guanidines like A-11, but where the ring A bearing NH is instead NR4 (where R4 is for example alkyl) can be used to make the compounds of formula I where R4 is other than H.
[000338] Thus, one embodiment is a method of making compounds of formula A-6, where the variables are defined as with respect to formula I, including: (i) reacting a guanidine of formula A-11 with a 1, 3-dielectrophile to make a 4-hydroxy-pyrimidin-2-yl-amine of the formula A-13; (ii) converting A-13 to a 4-pyrimidin-2-yl-amine leaving group of the formula A-14; and (iii) reacting the 4-leaving group-pyrimidin-2-yl-amine A-14 with an aryl or heteroaryl amine, A-3.
[000339] In one embodiment the 1,3-dielectrophil is an ester of β-aldehyde, A-12. In one embodiment, the leaving group of A-14 is a halo, in a more specific embodiment a chloro group. In one embodiment, the heteroaryl amine A-3, is a 5-aminobenzo[d]oxazol-2(3H)-one, substituted with groups according to formula I. In another embodiment, the heteroaryl amine A-3 is a 6-aminobenzo[d]oxazol-2(3H)-one, substituted with groups according to formula I. In another embodiment, heteroaryl amine A-3 is a 5-amino-1H - benzo[d]imidazole-2(3H)-one, substituted with groups according to formula I. EXAMPLES
[000340] The invention is further understood by reference to the following examples, which are not intended to be limiting. Any of the synthetic methods that are functionally equivalent are within the scope of the invention. Various modifications of the embodiments described herein would be apparent to a person of skill in the art from the foregoing description and accompanying figures. Such modifications fall within the scope of the appended claims.
[000341] One embodiment is a compound, according to formula I, as described in the examples below.
[000342] In the examples below as well as throughout the application, the following abbreviations have the following meanings. If not defined, terms have their generally accepted meanings.


[000343] The anilines used to manufacture the compounds described herein can be purchased commercially and/or synthesized by the methods disclosed herein and by those of skill in the art. Schemes (III) to (VII) below describe how some of the anilines were synthesized.
[000344] Scheme (III) describes a general synthetic route to anilines of formula A-16, where the group "OR" is used as an abbreviation for R2, where R2 is -ORe substituted with one or more of the same or different Ra and/or Rb as defined in relation to formula I. First, 3-fluorocatechol, is alkylated with the appropriate alkylating agent, for example an alkyl halide in the presence of a base, for example K2CO3 (typically 2.5 eq. of alkyl halide and base). In one example, the mixture is heated in acetone under reflux overnight. Half of the solvent is removed under reduced pressure and the salts are filtered off. The remaining solvent is removed in vacuo and the crude product is further purified by flash chromatography eluting with hexanes/ethyl acetate (0% ethyl acetate to 15% ethyl acetate gradient) to provide intermediate A-16.
[000345] Product A-16 is dissolved in dry THF and the vial is sealed with a rubber septum. N,N,N',N',N"-pentamethyldiethylenetriamine (PMDTA, 1.2 eq) is added via syringe and the vial is cooled to -75°C using an acetone/dry ice bath. n-Butyllithium (1.2 eq) is added via syringe and the reaction mixture is stirred for 90 minutes. The reaction mixture is subsequently poured into a beaker containing crushed dry ice and allowed to warm to room temperature. A 2M NaOH solution is added to the crude product and the mixture is washed with Et2O (2x) and EtOAc (1x). The aqueous phase is neutralized using HCl. The precipitated product, A-17, is collected by filtration and dried under vacuum. Scheme (III)

[000346] The carboxylic acid A-17 is combined azide (DPPA, 1.3 eq), triethylamine (1.3 eq) and dissolved in toluene. The resulting clear solution is refluxed for six hours and then concentrated under reduced pressure to obtain a Curtius Rearrangement product. The crude product, A-18, is further purified by flash chromatography eluting with dichloromethane/MeOH (0% to 5% MeOH gradient).
[000347] Urea A-18 is placed in a pressure flask together with a solution of NH4OH and NaBH4 (5.0 eq). The vessel is sealed and heated to 140°C for two days. The solvent is removed under reduced pressure and the crude product is purified using flash chromatography eluting with CHCl3/MeOH [with 2M NH3] (0% to 5% gradient) to furnish the aniline A-19. This procedure was used to make the anilines (ring A) for compounds I-50, I-77, I-101 and I-102.
[000348] Scheme (IV) describes a general synthetic route to anilines of formula A-25, where the group "R" is used as an abbreviation for R2, where R2 is -Re substituted with one or more of the same or different Ra and/or Rb as defined in relation to formula I. In an analogous manner as described above in relation to Scheme (III), ortho-fluoro phenol was alkylated to provide intermediates A-20. Ortho-lithiation and carbon dioxide quenching gave A-21 acids. The Curtius rearrangement provided the corresponding anilines, A-22, which were released from the urea carbonyl using aminolysis as described above to provide the anilines, A-23. Anilines A-23 have been used to manufacture, for example, compounds I-19 through I23. Scheme (IV)

[000349] Anilines A-23 were also brominated using N-bromosuccinimide (NBS), dissolving them in MeCN, adding NBS (1.0 eq) and stirring the reaction mixture for two hours at room temperature. Bromination occurs regio-selectively for the NH2 group. The solvent is removed under reduced pressure and the crude product is purified by column chromatography eluting with hexanes/ethyl acetate (9/1) to give the anilines A-24. Anilines A-24 were used, for example, to make compound I-27.
[000350] Anilines A-24 were also subjected to Kumada Bonding by dissolving them in THF and adding the solution to a pressure vessel. The desired alkyl magnesium halide (3M in THF, 3.3 eq) is added dropwise via syringe. Since the first two AlkylMgX equivalents abstract protons from aniline, gas evolution and heating are observed. After the addition of the Grignard reagent, the Pd-catalyst ([1,1'-Bis(diphenylphosphino)ferrocene]-dichloropalladium(II), 5% by mol) is added in small portions. The vial is sealed and heated to 80°C overnight. to quench the reaction, water is added very carefully. The mixture is subsequently extracted with EtOAc (3x) and the combined organic layers are passed through a MgSO4 buffer to remove residual water. After evaporation of the solvents the crude product is further purified by flash chromatography eluting with CHCl3/MeOH [2M NH3] (0% to 5% gradient) to give the anilines, A-25. Anilines A-25 were used to make compounds I-29, I-53 and I-68.
[000351] Scheme (V) illustrates another example of using a Kumada bonding reaction to install alkyl groups onto an aniline ring. specifically, Scheme (V) describes the synthesis of 3,4(d3),5-trimethylaniline. First, 3,5-dimethyl aniline (10.0 g, 82.5 mmol) was dissolved in MeCN. NBS (15.4 g, 86.6 mmol) was added and the reaction mixture was stirred for two days at room temperature. Analysis by TLC and HPLC indicated that in addition to the desired product several by-products were formed (regio-isomers and di-brominated by-products). Silica gel was added to the reaction mixture and the solvent was evaporated under reduced pressure. The resulting dry crude product was loaded onto a chromatography column and purified by elution with hexanes/ethyl acetate (gradient from 0% to 15% ethyl acetate). The product, 4-bromo-3,5-dimethylaniline, was obtained in 50% yield (8.21 g). MS (ES) 200/202 Scheme (V)

[000352] (M + H). The bromide (5.00 g, 24.9 mmol) was dissolved in dry THF (50 ml) and placed in a pressure vessel. Methyl-d3 magnesium chloride (82.2 ml, 82.2 mmol, 1 M in Bu2O) was added dropwise via syringe. During the addition of the first two equivalents a strong reaction, gas evolution and heating were observed as the aniline protons reacted with the Grignard reagent. After the addition was complete, the Pd catalyst ([1,1'-Bis(diphenylphosphino)ferrocene]-dichloropalladium(II), 0.911 g, 5% by mol) was added in small portions. The vial was sealed and heated to 80°C overnight. The reaction was carefully quenched with water. The mixture is subsequently extracted with EtOAc (3x) and the combined organic layers were passed through a MgSO4 plug to remove residual water. After evaporation of solvents the crude product was purified by flash chromatography eluting with DCM/MeOH (0% to 1% MeOH gradient). The 4-perterated 3,4,5-trimethylaniline was obtained in 43% yield (1.48 g). 1H NMR (300 MHz, DMSO) δ 6.19 (s, 2H), 4.57 (s, 2H), 2.05 (s, 6H) ppm; MS (ES) 139 (M + H). Compound I-35 was manufactured using this aniline.
[000353] Scheme (VI) shows the synthesis of 4-amino-2,6-dimethylbenzonitrile. First, 3.5 dimethyl-4-iodo aniline (5.0 g, 20.2 mmol), Cu(I)CN (2.17 g, 24.3 mmol) and DMF (80 ml) were placed in a vessel depression. The sealed bottle was heated in an oil bath to 185°C. After 10 minutes the bottle was removed from the oil bath and cooled to room temperature. LCMS analysis of the reaction mixture indicated a peak-to-peak reaction. Most of the DMF was removed under high vacuum using a rotary evaporator. The remaining crude product was loaded onto a scintillating column chromatography and purified by eluting with CHCl3/MeOH [2M NH3] (0% to 2% gradient). The product was obtained in 62% yield (2.21 g). 1H NMR (300 MHz, DMSO) δ 6.29 (s, 2H), 5.92 (br s, 2H), 2.23 (s, 6H) ppm; MS (ES) 147 (M + H). Compound I-64 was manufactured using this aniline. Scheme (VI)

[000354] Scheme (VII) describes the synthesis of 4-fluoro-3-methoxy-5-methylaniline. First, at 0°C, in a CH2Cl2 solution (150 ml) of 2-fluoro-3-methylphenol (1.12.6 g, 100 mmol) and HOAc (12 ml, 210 mmol), a solution in CH2Cl2 ( 50 ml) of bromine (10.5 ml, 205 mmol) was added dropwise in 45 minutes. After 15 minutes, the reaction was monitored to completion by LCMS, and was quenched by the addition of H2O (200 mL). Two layers were mixed well and then separated, the aqueous layer was extracted with CH 2 Cl 2 (50 ml), the organic layers were combined, dried (MgSO 4 ), filtered, the solvent removed in vacuo. 4,6-Dibromo-2-fluoro-3-methylphenol was obtained as a solid: 28.4 g (99% yield); 1H NMR (300 MHz, CDCl3) δ 7.48 (d, J = 2.1, 1H), 5.49 (d, J = 2.8, 1H), 2.30 (d, J = 2 .8, 3H); LRMS (M-) m/z 282.94.
[000355] To a homogeneous solution in CH3CN (200 ml) of 4,6-di-bromo-2-fluoro-3-methylphenol (2, 28.4 g, 100 mmol) at room temperature K2CO3 (16.59 g, 120 mmol) was added, followed by MeI (9.3 ml, 150 mmol). The cloudy solution was stirred at 30°C overnight. The reaction went to completion monitored by LC and TLC. The mixture was cooled to room temperature, quenched by the addition of H2O (100 ml) and sat. here (100 ml), two layers were mixed well then separated, the aqueous layer was extracted with EtOAc (50 ml), the organic layers were combined, dried (MgSO4 ), filtered, the solvent removed in vacuo. The crude product, an oil, was dissolved in CH2Cl2 (~80 ml) and the solution was passed through a silica gel pad (~50 g silica gel), washing with CH2Cl2 (~100 ml). The filtrate was collected, and the solvent was removed in vacuo. 1,5-Dibromo-3-fluoro-2-methoxy-4-methyl-benzene was obtained as a solid: 31.62 g (>99% yield); 1H NMR (300 MHz, CDCl 3 ) δ 7.53 (d, J = 2.1, 1H), 3.92 (s, 3H), 2.29 (d, J = 2.8, 3H). Scheme (VII)

[000356] Referring to Scheme VII, at 0°C, in a CH 2 Cl 2 (600 ml) solution of 2-fluoro-3-methylphenol (commercially available from Wonda Science, Montreal, Canada) (100 g, 792.8 mmol ) and HOAc (95 ml, 1.66 mol), a CH 2 Cl 2 (200 ml) solution of bromine (83 ml, 1.625 mol) was added dropwise in 2 hours. The reaction was >99% complete by LC-MS, and was quenched by the addition of H2O (500 ml). Two layers were mixed well and then separated, the aqueous layer was extracted with CH 2 Cl 2 (200 ml), the organic layers were combined, dried (MgSO 4 ), filtered, the solvent removed in vacuo. 4,6-Dibromo-2-fluoro-3-methylphenol was obtained as a yellowish white solid: 247.96 g (>99% yield); 1H NMR (300 MHz, CDCl3) δ 7.48 (d, J = 2.1, 1H), 5.42 (br s, 1H), 2.31 (d, J = 2.8, 3H) ; LRMS (M-) m/z 282.92.
[000357] 1,5-dibromo-3-fluoro-2-methoxy-4-methylbenzene was prepared from a homogeneous solution in CH3CN (1 L) of 4,6-dibromo-2-fluoro-3-methylphenol (~ 225.1 g, 792.8 mmol) by the addition at room temperature of K2CO3 (131.5 g, 1.18 mol) followed by MeI (74 ml, 951 mmol). The cloudy solution was stirred at 30°C and, as monitored by LC, the reaction went to completion at 21 hours. The reaction mixture was cooled to room temperature, the solid was filtered off, washed with CH2 Cl2 (~100 ml x 2). The filtrate was collected, the solvent was removed in vacuo. The crude product was suspended in CH 2 Cl 2 (200 ml) and the solution was passed through a silica gel pad (~50 g silica gel) (note, to remove some of the darker colored baseline impurities and the solid initially dissolved in CH3CN), washed with CH2Cl2 (~100 ml x 3). The filtrate was collected, the solvent was removed in vacuo. 1,5-Dibromo-3-fluoro-2-methoxy-4-methylbenzene was obtained as a yellowish white solid: 249.46 g (>99% yield); 1H NMR (300 MHz, CDCl 3 ) δ 7.53 (d, J = 2.1, 1H), 3.92 (s, 3H), 2.29 (d, J = 2.8, 3H).
[000358] To a suspension in H 2 SO 4 (100 ml) of 1,5-dibromo-3-fluoro-2-methoxy-4-methylbenzene (3.30 g, 100 mmol), HNO 3 (90% aq, 5.0 ml, 110 mmol) was added in 10 minutes, with the occasional ice bath cooling. Stirring was continued at 30°C for another 30 minutes. The reaction went to completion as monitored by the LC. The thick pasty mixture was cooled to room temperature and poured onto ice (~600 ml solid volume). A precipitate was collected by filtration, washed with water (~100 ml, final aqueous volume: ~600 ml), and dried further under vacuum. Crude 1,3-dibromo-5-fluoro-4-methoxy-6-methyl-2-nitrobenzene was obtained as a yellow solid (33.7 g), and was stirred in 90 ml of EtOH at 50°C during night. The cloudy solution was then cooled to room temperature, the solid was collected by filtration, washed with ice-cold EtOH. A first crop of 1,3-dibromo-5-fluoro-4-methoxy-6-methyl-2-nitro-benzene was obtained as a pale yellow solid: 22.6 g; a second crop was obtained from the precursor liquid as a yellow solid: 3.90 g (77% combined yield); 1H NMR (300 MHz, CDCl 3 ) δ 4.00 (s, 3H), 2.39 (d, J = 3.0, 3H).
[000359] In a Parr Flask with 1,3-dibromo-5-fluoro-4-methoxy-6-methyl-2-nitrobenzene (4, 102.9 g, 300 mmol) and sodium carbonate (33.4 g) , 315 mmol), under N 2 atmosphere, MeOH (600 ml) was added, followed by Pd-C (10% on activated carbon, 50% moisture, 5.0 g). The Parr flask was placed on a Parr shaker under 30 to 45 psi (310 kPa) (207 to 310 kPa) of hydrogen. after 21 hours the reaction went to completion as monitored by LCMS. The solid was filtered off to a pad of celite, and washed with MeOH. The filtrate was collected and the solvent was removed in vacuo. A light beige solid was obtained and treated with EtOAc (200 ml) and H 2 O (200 ml), two layers were mixed well and then separated, the aqueous layer was extracted with EtOAc (150 ml). The organic layers were combined and added to a flask with 25 ml conc. aq., with agitation. The precipitated solid was collected by filtration, washed with EtOAc, and dried further under high vacuum. A fluffy white solid was obtained as 1st crop: 29.99 g as an HCl salt; more white ppt was collected from the filtrate as 2nd crop: 17.12 g as an HCl salt; 3rd crop was obtained from the filtrate after removal of solvent and the solid resuspended in EtOAc: yellowish white solid, 8.90 g as HCl salt. Combined 4-fluoro-3-methoxy-5-methylbenzenamine as the HCl salt: 56.01 g (97% yield); 1H NMR (300 MHz, CD3OD) δ 6.92 (dd, J = 6.9, 2.5, 1H), 6.82 (dd, J = 5.3, 2.5, 1H), 3 .90 (s, 3H), 2.30 (d, J = 2.4, 3H); LRMS (M+) m/z 156.21. Compounds I-58, I80, I-105 through I-107, I-114, I-118, I-127, I-181, I-190 and I-191 were made using this aniline.
[000360] 3-Methoxy-4,5-dimethylaniline was prepared in an analogous manner as described in relation to Scheme (VII), starting with 2,3-dimethylphenol (61.08 g, 500 mmol). On bromination reaction, 4,6-dibromo-2,3-dimethylphenol was obtained as a solid: 140.74 g (>99% yield); 1H NMR (300 MHz, CDCl 3 ) δ 7.52 (s, 1H), 5.50 (s, 1H), 2.33 (s, 3H), 2.28 (s, 3H); LRMS (M-) m/z 278.97. On the alkylation reaction, 1,5-dibromo-2-methoxy-3,4-dimethylbenzene was obtained as a pale yellow oil at room temperature and resulted in a yellow solid after cooling with dry ice: 144.97 g (98, 6% yield in 2 steps); 1H NMR (300 MHz, CDCl 3 ) δ 7.59 (s, 1H), 3.76 (s, 3H), 2.32 (s, 3H), 2.30 (s, 3H). On nitration, 1,3-dibromo-4-methoxy-5,6-dimethyl-2-nitrobenzene was obtained as a yellowish white solid: 58.3 g (35% combined yield from two cycles of crystallization); 1H NMR (300 MHz, CDCl 3 ) δ 3.81 (s, 3H), 2.42 (s, 3H), 2.37 (s, 3H). In the reduction reaction, 3-methoxy-4,5-dimethylbenzenamine was obtained as a white solid (HCl salt) in a combined yield of in three recrystallization runs of 24 g (77%): 1H NMR (300 MHz, DMSO) δ 10.04 (s, 3H), 6.81 (s, 1H), 6.77 (s, 1H), 3.81 (s, 3H), 2.27 (s, 3H), 2 .09 (s, 3H); LRMS (M+) m/z 152.20. Compounds I-52, I-59, I-81, I-108 through I-113, I-115, I-125 and I-195 were manufactured using this aniline.
Scheme (VIII) describes the synthesis of 4-chloro-3-methoxy-5-methylaniline analogues to those reported in Journal of Medicinal Chemistry, 44(12), 1866-1882; 2001. A suspension of 4-bromo-2-methoxy-6-methylaniline (1 g, 4.63 mmol) in 28% HCl (2 ml) was cooled in a dry ice bath. To this a cooled solution of sodium nitrite (323 mg, 4.68 mmol) in water (1 ml) was added dropwise with vigorous stirring. In a separate flask a solution of cuprous chloride (2.29 g, 23.1 mmol) in concentrated HCl (2 ml) was prepared which was also cooled in a dry ice bath. The aniline mixture was quickly poured into this reaction vessel. The vial was allowed to warm to room temperature overnight. An additional 1 ml of 28% HCl was added and the reaction heated to 60°C for 3 hours monitored by LCMS. On cooling to room temperature, the reaction was diluted with water and extracted three times with ethyl acetate. The combined organic layers were dried over magnesium sulfate. Crude 5-bromo-2-chloro-1-methoxy-3-methylbenzene was purified by silica gel chromatography (0 to 20% ethyl acetate/hexanes) to yield 0.9 g (83%) of yellow solid clear 3. 1H NMR (300 MHz, CDCl 3 ) δ 7.01 (s, 1H), 6.91 (s, 1H), 3.88 (s, 3H), 2.35 (s, 3H).
[000362] The catalyst, (CyPF-t-Bu)PdCl2, was prepared according to Q. Shen and JF Hartwig in J. Am. Chem. Soc. 2006, 128(31), including supplementary information 10028-29. In one vial were combined 5-bromo-2-chloro-1-methoxy-3-methylbenzene (235 mg, 1.0 mmol), lithium amide (95%, 230 mg, 10 mmol), and (CyPF-t- Bu)PdCl2 (7.3 mg, 0.01 mmol) in 1,2-dimethoxyethane (2 ml) purged under argon. The reaction was heated to 80°C for 24 hours. On cooling to room temperature, the reaction was poured into ice water (2 ml), 1 M HCl (1 ml) was added, and stirred for 5 minutes. This was then neutralized with saturated NaHCO3 solution and extracted 3 x 10 ml with dichloromethane. Organic layers dried over MgSO4 and purified by silica gel chromatography (0 to 100% ethyl acetate/hexanes). This produced 30 mg of 4-chloro-3-methoxy-5-methylaniline. 1H NMR (300 MHz, CDCl 3 ) δ 6.20 (s, 1H), 6.15 (s, 1H), 3.83 (s, 3H), 2.27 (s, 3H); LCMS (m/z): 172 (MH+). Compound I-124 was manufactured using this aniline. Scheme (VIII)

[000363] Scheme (IX) describes the synthesis of 3-fluoro-5-methoxy-4-methylaniline. In a round bottom flask a solution of 1-chloro-4-fluoro-2-methoxybenzene (500 mg, 3.11 mmol) dissolved in anhydrous THF (10 ml) was prepared. On cooling to -78°C, N,N,N',N',N” - pentamethyldiethylenetriamine (PMDTA, 715 µl, 3.42 mmol) was added. After 20 minutes, n-butyllithium was added dropwise keeping the internal temperature at -78°C. After 3 hours, iodomethane was added, the cooling bath removed, and the reaction allowed to warm to room temperature overnight. TLC in 1:1 ethyl acetate/hexanes showed no starting material. The solvent volume was reduced via rotary evaporation, then the reaction was diluted with water and ethyl acetate. The organic phase was washed twice with 1N HCl, once with saturated NaHCO3 solution, once with brine, then dried over Na2SO4, and concentrated. This yielded 474 mg (87%) of 1-chloro-4-fluoro-2-methoxy-3-methylbenzene as a colorless oil. 1H NMR (300 MHz, CDCl3) δ 7.16 (dd, J = 8.9, 5.9 Hz, 1 H), 6.77 (t, J = 8.7 Hz, 1 H), 3.83 (s, 3H), 2.23 (s, 3H).
[000364] To a cooled to -78°C solution of 1-chloro-4-fluoro-2-methoxy-3-methylbenzene (470 mg, 2.69 mmol) in THF (5.5 ml) was added sodium amide (95%, 276 mg, 6.72 mmol). After 30 minutes, the solution was warmed to -20 to -30°C and then benzylamine (441 µl, 4.05 mmol) was added dropwise within 1 minute. The reaction was stirred overnight, and LCMS showed no starting material. The reaction was quenched by cooling in an ice bath followed by dropwise addition of saturated NH4Cl and water. After stirring for 1 hour, the reaction mixture was extracted twice with ethyl acetate, and the combined organic phases dried over MgSO4 and concentrated by rotary evaporation. The crude product was purified by silica gel chromatography (0 to 30% ethyl acetate/hexanes) to yield 223 mg (34%) of N-benzyl-3-fluoro-5-methoxy-4-methylaniline as an oil Orange. 1H NMR (300 MHz, CDCl 3 ) δ 7.40 - 7.27 (m, 5H), 6.06 - 6.00 (m, 1H), 6.00 (s, 1H), 4.29 ( s, 2H), 3.74 (s, 3H), 2.00 (s, 3H); LCMS (m/z): 246 (MH+). Scheme (IX)

[000365] To a solution of N-benzyl-3-fluoro-5-methoxy-4-methylaniline (223 mg, 0.91 mmol) in methanol (25 ml) prepared in a Parr shaker flask was added palladium hydroxide ( 20% in carbon). This was subjected to 45 psi (310 kPa) of hydrogen gas overnight until LCMS showed complete conversion. The catalyst was filtered and the solution concentrated in vacuo to yield 117 mg (83%) of 3-fluoro-5-methoxy-4-methylaniline as an oil. 1H NMR (300 MHz, CDCl 3 ) δ 6.03 (d, J = 12.0 Hz, 1H), 6.01 (s, 1H), 3.77 (s, 3H), 2.09 (s , 3H); LCMS (m/z): 156 (MH+). Compound I-120 was manufactured using this aniline. Example 2: Synthesis of 2-halo-pyrimidine-4-amines
[000366] Synthesis of 5-(2-chloro-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one:
[000367] To a vial with 5-aminobenzo[d]oxazol-2(3H)-one (300.1 mg, 2.0 mmol) and 2,4-dichloro-5-methylpyrimidine (423.8 mg, 2, 6 mmol), MeOH (8 ml) and H2O (2 ml) were added. The turbid mixture was stirred at room temperature for 64 hours. The precipitate from the reaction mixture was collected by filtration, washed with EtOAc (3 ml x 2), and was further dried in vacuo. 5-(2-Chloro-5-methylpyrimidin-4-ylamino)benzo[d]-oxazol-2(3H)-one was obtained as a yellowish white solid: 394 mg (71% yield); 1H NMR (300 MHz, DMSO) δ 11.68 (br s, 1H), 8.62 (s, 1H), 7.94 (d, J = 0.8, 1H), 6.97 ( d, J = 2.0, 1 H), 6.82 (d, J = 8.1, 1 H), 6.74 (dd, J = 2.0, 8.1, 1 H), 2, 15 (s, 3H); LCMS (M+) m/z 277.10.
[000368] Synthesis of 5-(2-chloro-5-fluoropyrimidin-4-ylamino)-1H-benzo[d]imidazol-2(3H)-one:
[000369] To a vial with 5-amino-1H-benzo[d]imidazol-2(3H)-one (298.3 mg, 2.0 mmol) and 2,4-dichloro-5-fluoropyrimidine (434.1 mg, 2.6 mmol), MeOH (8 ml) and H2O (2 ml) were added. The turbine solution was stirred at room temperature for 3 days. The precipitate from the reaction mixture was collected by filtration, and washed with EtOAc (3 ml x 2), and was further dried in vacuo. 5-(2-Chloro-5-fluoropyrimidin-4-ylamino)-1H-benzo-[d]imidazol-2(3H)-one was obtained as a yellowish white solid: 390.3 mg (70% yield) ; 1H NMR (300 MHz, DMSO) δ 10.69 (s, 1H), 10.63 (s, 1H), 9.87 (s, 1H), 8.27 (d, J = 3.6 , 1H), 7.35 (d, J = 1.9, 1H), 7.18 (dd, J = 1.9, 8.3, 1H), 6.93 (d, J = 8 .3, 1H); LCMS (M+) m/z 279.80.
[000370] Synthesis of 6-(2-chloro-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one:
[000371] To a vial with 6-aminobenzo[d]oxazol-2(3H)-one (1.0 g, 6.7 mmol) and 2,4-dichloro-5-methylpyrimidine (1.4 g, 8, 7 mmol), solvents MeOH (20 ml) and H2O (5 ml) were added. The turbid mixture was stirred at room temperature for 2 days. The precipitate from the reaction mixture was collected by filtration, washed with H 2 O (3 ml x 2) and EtOAc (3 ml x 2), and dried further in vacuo. 6-(2-Chloro-5-methylpyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one was obtained as a light brown solid: 1.59 g (86% yield); 1H NMR (300 MHz, DMSO) δ 11.59 (s, 1H), 8.87 (s, 1H), 7.99 (s, 1H), 7.56 (s, 1H), 7 .28 (d, J = 8.3, 1H), 7.06 (d, J = 8.3, 1H), 2.14 (s, 3H). Example 3: Synthesis of 2,4-pyrimidinediamines using 2-halo-pyrimidine-4-amines.

[000372] 5-Fluoro-N2-(3,4,5-trimethyl)phenyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine
[000373] 2-Chloro-5-fluoro-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-4-pyrimidineamine (50 mg) and 3,4,5-trimethylaniline (50 mg) were suspended in isopropanol (1 ml) and TFA (5 drops). The solution was heated at 100°C overnight in a sealed tube, then cooled to room temperature. LCMS showed complete product conversion. The reaction solution was diluted with 2.0 M NH3 in methanol (5 ml). The solution was sonicated. The precipitation was filtered off and washed with methanol (10 ml), dried to give the desired product. 1H NMR (300 MHz, DMSO) δ 9.98 (br, 1H), 9.27 (s, 1H), 8.92 (s, 1H), 8.04 (d, J = 3.6 , 1H), 7.42 (d, J = 8.7, 1H), 7.33 (s, 1H), 7.23 (s, 2H), 7.18 (d, J = 8. 7.1H), 2.05 (s, 6H), 2.00 (s, 3H); 19F NMR (282 MHz, DMSO) δ - 181.06; LCMS: purity: 100%; MS (m/e): 380.13 (MH+). N2-(3,4,5-trimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine

[000374] 2-Chloro-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-4-pyrimidineamine (300 mg) and 3,4,5-trimethylaniline (300 mg) were suspended in isopropanol (3 ml) and TFA (10 drops). The solution was heated with microwaves at 160°C for 30 minutes in a sealed tube, then cooled to room temperature. LCMS showed complete product conversion. The reaction solution was diluted with 2.0 M NH3 in methanol (5 ml). The solution was sonicated. The precipitation was filtered off and washed with methanol (10 ml), dried to give the desired product. 1H NMR (300 MHz, DMSO) δ 11.39 (br, 1H), 8.70 (s, 1H), 8.29 (s, 1H), 7.83 (s, 1H), 7 .30 (d, J = 6.3, 2H), 7.21 (m, 3H), 2.06 (s, 3H), 1.99 (s, 6H), 1.98 (s, 3H); LCMS: purity: 96.73%; MS (m/e): 376.27 (MH+).

[000375] N2-(2-methoxypyridin-4-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine-chloro -5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-4-pyrimidineamine (50 mg), 2-methoxy-4-aminopyridine (50 mg), acetate of palladium (30 mg), BINAP (30 mg) and cesium carbonate (30 mg) were suspended in dioxane (1 ml) and NMP (0.5 ml). The reaction mixture was heated with microwaves at 180°C for 30 minutes in a sealed tube, then cooled to room temperature. The reaction solution was diluted with methanol (5 ml) and sonicated. It was filtered through celite and washed with methanol. The filtrate was evaporated and purified by HPLC to give the desired product. 1H NMR (300 MHz, DMSO) δ 11.65 (s, 1H), 10.16 (br, 1H), 9.11 (br, 1H), 7.98 (s, 1H), 7 .89 (d, J = 6.0Hz, 1H), 7.31 - 7.10 (m, 5H), 3.72 (s, 3H), 2.14 (s, 3H); LCMS: purity: 97.39%; MS (m/e): 365.35 (MH+). 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one

[000376] In a three-necked round bottom flask, 5-(2-chloro-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one (33.2 g, 120 mmol) and 4-fluoro-3-methoxy-5-methylbenzenamine hydrochloride (32.19 g, 168 mmol) was added, followed by i-PrOH (750 ml) and TFA (23.1 ml, 300 mmol). The reaction mixture was stirred under nitrogen atmosphere with gentle reflux (note: with overhead stirrer at internal temperature 75 to 80°C). After 22 hours, by LC-MS, the reaction was >95% complete (note: when <3% of the 2-chloropyrimidine starting material remained, the reaction did not progress much further even after another day or two). The reaction mixture was cooled to room temperature, the precipitate was collected by filtration, washed with i-PrOH (~50 ml x 2). 5-(2-(4-Fluoro-3-methoxy-5-methylphenyl-amino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one Trifluoroacetate Compound was obtained as a light brown solid (still slightly moist) and added to an aqueous solution of NaHCO3 (15.12 g (180 mmol) of NaHCO3 dissolved in 1 liter of H2O). The aqueous suspension was stirred at room temperature overnight. The solid was collected by filtration, washed with H2O (~150 ml x 3), and dried further under high vacuum. The free base compound 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)benzo-[d]oxazol-2(3H)-one was obtained as a solid yellowish white: 44.51 g (93.8% yield).
[000377] The benzenesulfonic salt (besylate) of the illustrated product was obtained as follows: In a suspension of MeOH (400 ml) of 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin- 4-ylamino)benzo[d]-oxazol-2(3H)-one (free base; 39.5 g, 100 mmol), with stirring, benzenesulfonic acid (16.61 g, 105 mmol) was added (note: a reaction mixture turned to an almost homogeneous light brown solution for a short time, ppts appeared shortly thereafter). The cloudy solution was stirred at 50°C for 90 minutes, cooled to room temperature, solid was collected by filtration, washed with EtOAc (~150 ml x 2), and dried further under high vacuum. 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one besylate Compound was obtained as a very light brown solid: 52.09 g (94% yield); 1H NMR (300 MHz, DMSO) δ 11.99 (br s, 1H), 11.80 (s, 1H), 10.05 (s, 1H), 9.81 (s, 1H), 7.87 (s, 1H), 7.65 - 7.62 (m, 2H), 7.39 - 7.33 (m, 4H), 7.29 - 7.26 (m, 2H), 6 .96 (br d, J = 7.4 Hz, 1 H), 6.91 (br d, J = 5.6 Hz, 1 H), 3.66 (s, 3H), 2.20 (s, 3H), 2.06 (s, 3H); LRMS (M+) m/z 396.15 Example 4: Synthesis of 2,4-pyrimidinediamines where the pyrimidine nucleus is installed via guanyl A ring analogues.
[000378] Scheme (X) describes the synthesis of N2-(3,5-dimethyl-4-fluoro)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5 -yl)-2,4-pyrimidinediamine starting from 3,5-dimethyl-4-fluoroaniline. First, 3,5-dimethyl-4-fluoroaniline (500 mg), N,N-bis-Boc-1-guanylpyrazole (1.7 g, 1.5 eq.) and triethylamine (0.75 ml, 1.5 eq.) were dissolved in anhydrous THF (10 ml). The reaction solution was heated at 50°C for two days, then evaporated to dryness. The residue was dissolved in dichloromethane (1 ml), then trifluoroacetic acid was added to the solution (9 ml). The solution was stirred at room temperature overnight. LCMS confirmed guanidine formation. The solution was evaporated and the residue recrystallized from ethyl acetate and hexanes to give a beige solid as the TFA salt of 3,5-dimethyl-4-fluorophenylguanidine (740 mg, 70%). 1H NMR (300 MHz, DMSO) δ 9.48 (s, 1H), 7.58 (s, 1H), 7.28 (s, 3H), 6.96 (d, J = 6.3, 2H), 2.20 (s, 6H); 19F NMR (282 MHz, DMSO) δ - 139.45.
3,5-Dimethyl-4-fluorophenylguanidine TFA salt (700 mg) and acid ethyl ester (724 mg) were dissolved in anhydrous ethanol (10 ml). to this solution, sodium ethoxide (1.18 g) was added. The reaction solution was heated at 70°C for two days. LCMS showed 10% guanidine starting material remaining. The solution was cooled to room temperature and diluted with water (100 ml). The mixture was extracted with ethyl acetate (3 x 100 ml) and evaporated. The residue was purified by column chromatography (methanol in dichloromethane = 0 to 30% in 35 minutes) to give N2-(3,5-dimethyl-4-fluoro)phenyl-4-hydroxy-5-methyl-2-pyrimidineamine (320mg, 60%). 1H NMR (300 MHz, DMSO) δ 10.76 (br, 1H), 8.42 (s, 1H), 7.57 (s, 1H), 7.23 (d, J = 6.0 , 2H), 2.16 (s, 6H), 1.79 (s, 3H); LCMS: purity: 93.45%; MS (m/e): 248.07 (MH+). Scheme (X)

[000380] N2-(3,5-dimethyl-4-fluoro)phenyl-4-hydroxy-5-methyl-2-pyrimidineamine (120 mg) was suspended in anhydrous THF (2 ml). Then POCl3 (0.2 ml) was added to the mixture. The solution was heated to 60°C for one hour. LCMS showed complete conversion of the 4-hydroxy starting material to the corresponding 4-chloro product. The reaction was quenched with water (40 ml) and extracted with ethyl acetate (40 ml). The organic layer was evaporated and purified by column chromatography (EtOAc in hexanes = 0 to 60% in 35 minutes) to give 4-chloro-N2-(3,5-dimethyl-4-fluoro)phenyl-5-methyl-2 -pyrimidine-amine (100 mg, 77%). 1H NMR (300 MHz, DMSO) δ 9.69 (s, 1H), 8.63 (s, 1H), 7.34 (d, J = 6.6, 2H), 2.16 (s, 6H), 2.14 (s, 3H); 19F NMR (282 MHz, DMSO) δ - 145.51; LCMS: purity: 97.40%; MS (m/e): 266.06 (MH+).
[000381] 4-Chloro-N2-(3,5-dimethyl-4-fluoro)phenyl-5-methyl-2-pyrimidine-amine (80 mg) and 5-amino-2(3H)-benzoxazolone (80 mg) were dissolved in isopropanol (1 ml) and trifluoroacetic acid (5 drops). The solution was heated at 75°C overnight and then diluted with 2.0 M ammonia in methanol (10 ml). The reaction mixture was sonicated and the precipitation was filtered off, washed with methanol, dried to give N2-(3,5-dimethyl-4-fluoro)phenyl-5-methyl-N4-(2-oxo-2,3 -dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine. 1H NMR (300 MHz, DMSO) δ 11.59 (s, 1H), 8.86 (s, 1H), 8.33 (s, 1H), 7.84 (s, 1H), 7 .27 (m, 4H), 7.22 (d, J = 9.0, 1H), 2.06 (s, 3H), 1.98 (s, 6H); 19F NMR (282 MHz, DMSO) δ - 147.88; LCMS: purity: 99.86%; MS (m/e): 380.21 (MH+). The free base compound was converted to a besylate salt, compound I-146, and a sulfate salt, compound I-151. Example 5: Synthesis of Exemplary Prodrug Compounds
[000382] The exemplary drug compounds disclosed herein were synthesized as illustrated in Scheme (XI) Scheme (XI)

[000383] Referring to Scheme (XI), a suspension in DMF of 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d] trifluoroacetate oxazol-2(3H)-one (prepared as described in Example 3) (40.38 g, 79 mmol) and Cs2CO3 (78.3 g, 238 mmol) was stirred at room temperature under a nitrogen atmosphere, after 2, 5 hours, di-tert-butyl chloromethyl phosphate (26.6 g, 103 mmol) was added and stirring was continued at room temperature. After 49 hours, by LC-MS, the reaction was >90% complete (note: due to the insufficient quality of the alkylating agent from this batch, an additional ~7 g of di-tert-butyl chloromethyl phosphate was added). With stirring, the reaction mixture was poured into 1.2 liter of H2O, ppt formed and was somewhat lumpy, an additional 500 ml of H2O was added to ensure formation of the non-sticky solid. The precipitate was collected by filtration, washed with water, and dried further under high vacuum. The compound (5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)-2-oxobenzo[d]-oxazol-3(2H)-yl)methyl phosphate di-tert-butyl was obtained as a light brown solid and was used directly in the next reaction.
[000384] Ao (5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)-2-oxobenzo[d]oxazol-3(2H)-yl) crude di-tert-butyl methyl phosphate, HOAc (280 ml) and H2O (70 ml) were added, the resulting pale brown homogeneous solution was stirred at 65°C (note: a lighter colored ppt was formed between 15 to 45 minutes). The reaction was monitored by LC-MS. After 1 hour the mixture was cooled to room temperature, the ppt was collected by filtration, washed with H2O (~50 ml x 2). The dihydrogen phosphate product of (5-(2-(4-fluoro-3-methoxy-5-methylphenyl-amino)-5-methylpyrimidin-4-ylamino)-2-oxobenzo[d]oxazol-3(2H)- il)methyl was obtained as a light pinkish beige solid and was used directly in salt formation.
[000385] With stirring, to a suspension of H2O (500 ml) of (5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)-dihydrogen phosphate Crude 2-oxobenzo[d]oxazol-3(2H)-yl)methyl, 1N NaOH (aq., 163 ml, 163 mmol) was added dropwise in 15 minutes, with cooling in an ice bath. remove ice bath, add more H2O until final volume reaches ~1 liter. The solid was removed by filtration through a filter paper, washed with H2O. The solid (~3 g) appeared as a 1:1 mixture of prodrug and precursor compound (I-105) which was formed during the previous hydrolysis of the reaction.). The filtrate was collected, and more H2O was removed by lyophilization (~20 mL of H2O left as solid ice). With stirring, i-PrOH (800 ml) was added to the solid, stirring was continued for 1 hour until the solid was evenly dispersed in the solution. The solid was collected by filtration, washed with i-PrOH (~100 ml x 3), and dried further under high vacuum. The compound of (5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)-2-oxobenzo[d]oxazol-3(2H)-yl)methyl phosphate disodium was obtained as a light beige solid: 38.87 g (89% yield over 3 steps, starting from the TFA salt described above); 1H NMR (300 MHz, D2O) δ 7.63 (s, 1H), 7.39 - 7.33 (m, 2H), 7.13 - 7.09 (m, 1H), 6.77 - 6.71 (m, 2H), 5.41 (d, J = 6.1 Hz, 2H), 3.62 (s, 3H), 2.01 (br s, 3H), 1.99 (br s , 3H); LRMS (M-) m/z 504.12. Example 5: Exemplary Synthesized Compounds
[000386] The following compounds were manufactured in a manner similar to the above examples or by methods described herein or known to a person of ordinary skill in the art. Analytical data and/or exemplary experimental procedure to manufacture selected compounds follow their name below. I-1: 5-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-2-methylbenzonitrile formate salt I-2: 4-(4-(2-Oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide Trifluoroacetate Salt I-3: 3-(4-(2) Trifluoroacetate Salt -oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide I-4: 5-(5-Chloro-2-(phenylamino)pyrimidin-4-ylamino) trifluoroacetate salt benzo[d]oxazol-2(3H)-one I-5: 5-(5-Chloro-2-(3,4-dimethylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2 trifluoroacetate salt (3H)-one I-6: 5-(5-Chloro-2-(3,4,5-trimethylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt I-7: 5-(5-Chloro-2-(2,4-difluoro-3-methoxyphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt I- 8: 5-(5-Chloro-2-(3-chloro-5-fluorophenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt I-9: 5-(5 -chloro-2-(4-methyl-3-(trifluoromethyl)phenylamino)pyrimidin-4-ylamino)-benzo[d ]oxazol-2(3H)-one I-10: 5-(5-Chloro-2-(3,5-dimethylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) trifluoroacetate salt -one I-11: 4-(5-Chloro-4-(2-oxo-2,3-dihydrobenzo[d]-oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide trifluoroacetate salt I-12: 5-(5-chloro-2-(3-methoxy-5-(trifluoromethyl)phenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-13: 5-(Trifluoroacetate) salt 2-(3,5-difluorophenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-14: 5-(5-Chloro-2-(3,5-difluorophenylamino) trifluoroacetate salt )-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-15: 5-(5-Chloro-2-(4-(trifluoromethyl)phenylamino)-pyrimidin-4-trifluoroacetate salt ylamino)benzo[d]oxazol-2(3H)-one I-16: 5-(5-Bromo-2-(3,4,5-trimethylphenylamino)pyrimidin-4-ylamino)benzo[d]formate salt oxazol-2(3H)-one I-17: 5-[2-(3-Dimethylamino-4-methyl-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one 1H NMR ( 300 MHz, DMSO) δ 11.57 (s, 1H), 8.32 (d, J = 13.0, 2H), 7.82 (s, 1H), 7.35 (dd, J = 8.3, 16.0, 3H), 7.10 (d, J = 8.7, 1H), 6.92 (t, J = 8.3, 1H), 6, 78 (t, J = 8.0, 1H), 3.79 (s, 3H), 3.34 (s, 2H), 2.07 (s, 3H) ppm; MS (ES) 391 (M + H). I-18: 5-[5-Ethynyl-2-(3,4,5-Trimethyl-phenylamino)-pyrimidin-4-ylamino]-3H-benzooxazol-2-one trifluoroacetate salt MS (ES) 386 (M + H). I-19: 5-[2-(2-Fluoro-3-methoxy-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one 1H NMR (300 MHz, DMSO) δ 11, 75 (s, 1H), 9.91 (s, 1H), 9.65 (s, 1H), 7.87 (s, 1H), 7.26 - 7.06 (m, 4H) , 6.99 (d, J = 5.6, 2H), 3.81 (s, 3H), 2.12 (s, 3H) ppm; MS (ES) 382 (M + H). I-20: 5-(5-Fluoro-2-(2-fluoro-3-methoxyphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt I-21: Salt of 5-(5-Chloro-2-(2-fluoro-3-methoxyphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate I-22: 5-Trifluoroacetate salt (5-bromo-2-(2-fluoro-3-methoxyphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-23: 5-(2-(2-fluoro-3) -methoxyphenylamino)pyrimidin-4-ylamino)benzo[d]-oxazol-2(3H)-one I-24: 5-[2-(3-Dimethylamino-phenylamino)-5-methyl-pyrimidin-4 trifluoroacetate salt -ylamino]-3H-benzooxazol-2-one 1H NMR (300 MHz, DMSO) δ 11.82 (s, 1H), 10.41 (s, 1H), 9.76 (s, 1H), 7.86 (s, 1H), 7.24 (s, 3H), 7.03 (t, J = 8.1, 1H), 6.73 (d, J = 8.1, 1H) , 6.66 (s, 1H), 6.48 (d, J = 8.2, 1H), 2.70 (s, 6H), 2.12 (s, 3H); ppm; MS (ES) 377 (M + H). I-25: 5-[2-(4-Dimethylamino-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one 1H NMR (300 MHz, DMSO) δ 11.63 (br br , 1H), 8.59 (s, 1H), 8.24 (s, 1H), 7.78 (s, 1H), 7.40 (d, J = 8.9, 2H), 7.34 (d, J = 7.9, 2H), 7.18 (d, J = 9.0, 1H), 6.57 (d, J = 9.0, 2H), 2.77 ( s, 6H), 2.05 (s, 3H) ppm; MS (ES) 377 (M + H). I-26: 5-[5-Methyl-2-(methyl-phenyl-amino)-pyrimidin-4-ylamino]-3H-benzo-oxazol-2-one 1H NMR (300 MHz, DMSO) δ 8.16 ( s, 1H), 7.76 (s, 1H), 7.43 - 7.22 (m, 5H), 7.13 (t, J = 7.1, 1H), 6.96 (d , J = 8.7, 1H), 3.37 (s, 3H), 2.03 (s, 3H) ppm; MS (ES) 348 (M + H). I-27: 5-[2-(4-Bromo-2-fluoro-3-methoxy-phenylamino)-5-methyl-pyrimidin-4-yl-amino]-3H-benzooxazol-2-one 1H NMR (300 MHz , DMSO) δ 8.39 (s, 1H), 8.26 (s, 1H), 7.81 (s, 1H), 7.66 (t, J = 8.5, 1H), 7.24 - 7.08 (m, 2H), 6.99 (d, J = 8.3, 1H), 3.77 (s, 2H), 2.06 (s, 2H) ppm; MS (ES) 460/462 (M + H). I-28: 5-[2-(4-Bromo-2-fluoro-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one 1H NMR (300 MHz, DMSO) δ 11, 80 (s, 1H), 10.11 (s, 1H), 9.69 (s, 1H), 7.91 (s, 1H), 7.63 (d, J = 10.3, 1H), 7.51 (t, J = 8.6, 1H), 7.38 - 7.02 (m, 4H), 2.12 (s, 3H) ppm; MS (ES) 430/432 (M + H). I-29: 5-[2-(2-Fluoro-3-methoxy-4-methyl-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one trifluoroacetate salt 1H NMR ( 300 MHz, DMSO) δ 11.79 (s, 1H), 10.21 (s, 1H), 9.81 (s, 1H), 7.88 (s, 1H), 7.18 ( s, 2H), 7.12 (d, J = 7.9, 1H), 6.92 (d, J = 8.4, 1H), 3.70 (s, 3H), 2.18 ( s, 3H), 2.13 (s, 3H) ppm; MS (ES) 396 (M + H). I-30: {4-[5-Methyl-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidin-2-ylamino]-phenyl}-acetaldehyde 1H NMR (300 MHz, DMSO δ 9.79 (s, 1H), 8.43 (s, 1H), 7.91 (s, 1H), 7.75 (q, J = 8.9, 3H), 7.28 (m, 2H), 2.51 - 2.37 (m, 2H), 2.10 (s, 3H) ppm; MS (ES) 376 (M + H). I-31: 5-[2-(3-Ethynyl-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzo-oxazol-2-one 1H NMR (300 MHz, DMSO) δ 9.10 ( s, 1H), 8.31 (s, 1H), 7.84 (d, J = 14.0, 1H), 7.68 (d, J = 9.3, 1H), 7. 28 (s, 1H), 7.20 (d, J = 8.6, 1H), 7.17 - 7.03 (m, 2H), 6.89 (d, J = 7.5, 1 H), 3.94 (s, 1H), 2.07 (s, 3H) ppm; MS (ES) 358 (M + H). I-32: 5-(5-Chloro-2-(3-methoxy-4-methylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt I-33: 5 -(5-chloro-2-(3-(dimethylamino)-4-methylphenylamino)pyrimidin-4-yl-amino)benzo[d]oxazol-2(3H)-one I-34: 5-[2-(3 -Amino-4-methoxy-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one 1H NMR (300 MHz, DMSO) δ 8.44 (s, 1H), 8.04 (s, 1H), 7.75 (s, 1H), 7.32 (s, 1H), 7.11 (d, J = 8.6, 1H), 7.03 (d, J = 2.2, 1H), 6.98 (d, J = 8.4, 1H), 6.71 (dd, J = 2.3, 8.6, 1H), 6.57 (d , J = 8.7, 1H), 3.66 (s, 3H), 2.04 (s, 3H) ppm; MS (ES) 379 (M + H). I-35: 5-[5-Methyl-2-(3,5-dimethyl-4-d3-methyl-phenylamino)-pyrimidin-4-ylamino]-3H-benzooxazol-2-one trifluoroacetate salt 1H NMR ( 300 MHz, DMSO) δ 11.84 (s, 1H), 10.36 (s, 1H), 9.89 (s, 1H), 7.90 (s, 1H), 7.32 ( d, J = 8.3, 1H), 7.22 (s, 1H), 6.97 (s, 2H), 2.13 (s, 3H), 1.99 (s, 6H) ppm; MS (ES) 379 (M + H). I-36: 5-(5-Chloro-2-(3,5-dimethyl-4-d3-methyl)phenyl-amino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) trifluoroacetate salt -one I-37: 5-(5-Fluoro-2-(3,5-dimethyl-4-d3-methyl)phenyl-amino)pyrimidin-4-ylamino)benzo[d]oxazol-2(trifluoroacetate salt) 3H)-one I-38: 5-(2-(3,5-dimethyl-4-d3-methyl)phenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-trifluoroacetate salt One I-39: 4-(2-Oxo-2,3-dihydro-benzooxazol-5-ylamino)-2-phenylamino-pyrimidine-5-carboxylic acid methyl ester trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.71 (s, 1H), 9.97 (d, J = 30.6, 2H), 8.70 (d, J = 3.4, 1H), 7.62 (s, 2H) , 7.43 - 7.08 (m, J = 20.8, 5H), 6.96 (s, 1H), 3.83 (s, J = 3.4, 3H), 3.40 (s , 2H) ppm; MS (ES) 378 (M + H). I-40: 4-(2-Oxo-2,3-dihydro-benzooxazol-5-ylamino)-2-(3,4,5-trimethyl-phenylamino)-pyrimidine-5-acid methyl ester trifluoroacetate salt carboxylic 1H NMR (300 MHz, DMSO) δ 11.75 (s, 1H), 10.13 (s, 1H), 9.82 (s, 1H), 8.22 (d, J=4, 8.1H), 7.49 - 7.19 (m, 3H), 7.12 (s, 2H), 6.95 (s, 1H), 3.75 (s, 3H), 2.24 (s, 3H), 2.05 (s, 6H) ppm; MS (ES) 420 (M + H). I-41: 5-(5-Nitro-2-(phenylamino)pyrimidin-4-yl-amino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt I-42: 5-trifluoroacetate salt (5-nitro-2-(3,4,5-trimethylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-43: 5-(2-(2) Trifluoroacetate salt ,4-Difluoro-3-methoxyphenylamino)-5-nitropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-44: 5-(2-(3-Methoxy-4-trifluoroacetate) salt methylphenylamino)-5-nitro-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-45: 5-(2-(3-(dimethylamino)-4-methylphenylamino)-trifluoroacetate salt 5-nitropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-46: 5-(5-methyl-2-(3,5-dimethyl-4-d3-methyl)phenylamino)pyrimidin- 4-ylamino)-benzo[d]oxazol-2(3H)-one I-47: 5-(2-(3-Methoxy-5-(trifluoromethyl)phenylamino)-5-nitropyrimidin-4-ylamino trifluoroacetate salt )benzo[d]oxazol-2(3H)-one I-48: 2-(2,4-Difluoro-3-methoxy-phenylamino)-4-(2-oxo-2, acid methyl ester trifluoroacetate salt, 3-dihydro-benzooxazol-5-ylamino)-pyrimidine-5-carboxylic MS (E S) 444 (M + H). I-49: 4-(2-Oxo-2,3-dihydro-benzooxazol-5-ylamino)-2-(3,4,5-trimethyl-phenylamino)-pyrimidine-5-carboxylic acid trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.68 (s, 1H), 10.33 (s, 1H), 9.67 (s, 1H), 8.65 (s, 1H), 7.71 - 7.49 (m, 1H), 7.36 - 7.21 (m, 2H), 3.42 (s, 12H), 2.48 (s, 5H), 2.02 (s, 3H) ppm; MS (ES) 406 (M + H). I-50: 5-[2-(2-Fluoro-3,4-dimethoxy-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one 1H NMR (300 MHz, DMSO) δ 8.17 (d, J = 5.3, 2H), 7.76 (s, 1H), 7.37 - 7.19 (m, 3H), 7.00 (d, J = 8.4, 1H), 6.74 (d, J = 9.2, 1H), 3.78 (s, 3H), 3.72 (s, 3H), 2.04 (s, 3H) ppm; MS (ES) 412 (M + H). I-51: 5-(5-Chloro-2-(2-fluoro-3,4-dimethoxyphenyl-amino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt I- 52: 5-(5-chloro-2-(3-methoxy-4,5-dimethylphenylamino)pyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one I-53: 5-{2- [2-Fluoro-3-(2-methoxy-ethoxy)-4-methyl-phenylamino]-5-methyl-pyrimidin-4-ylamino}-3H-benzooxazol-2-one 1H NMR (300 MHz, DMSO) δ 8 .27 (s, 1H), 8.21 (s, 1H), 7.80 (s, 1H), 7.46 - 7.25 (m, 3H), 7.09 (d, J = 8.6, 1H), 6.80 (d, J = 8.5, 1H), 4.01 - 3.93 (m, 2H), 3.57 - 3.52 (m, 2H), 3.27 (s, 3H), 2.16 (s, 3H), 2.05 (s, 3H) ppm; MS (ES) 440 (M + H). I-54: 2-(4-Carbamoyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine-5-carboxylic acid methyl ester trifluoroacetate salt 1H NMR ( 300 MHz, DMSO) δ 11.73 (s, 1H), 10.16 (s, 1H), 10.05 (s, 1H), 8.86 - 8.62 (m, 1H), 7.81 (s, 1H), 7.69 (m, 2H), 7.32 (m, 3H), 3.84 (s, 3H) ppm; MS (ES) 421 (M + H). I-55: 2-(3-Methoxy-5-trifluoromethyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine-5-acid methyl ester trifluoroacetate salt carboxylic 1H NMR (300 MHz, DMSO) δ 11.62 (s, 1H), 10.16 (s, 1H), 9.98 (s, 1H), 8.73 (s, 1H), 7.51 (s, 2H), 7.24 (d, J = 8.4, 2H), 7.20 - 7.03 (m, 1H), 6.77 (s, 1H), 3. 84 (s, 3H), 3.66 (s, 3H) ppm; MS (ES) 473 (M + H). I-56: 2-(3-Methoxy-4-methyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine-5-acid methyl ester trifluoroacetate salt carboxylic MS (ES) 422 (M + H). I-57: 2-(3-Dimethylamino-4-methyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine-5-acid methyl ester trifluoroacetate salt carboxylic 1H NMR (300 MHz, DMSO) δ 11.78 (s, 1H), 10.25 - 9.90 (m, 2H), 8.79 - 8.64 (m, 1H), 7.81 - 7.44 (m, 2H), 7.44 - 7.20 (m, 3H), 7.10 (s, 1H), 3.83 (s, 3H), 2.83 (s, 6H) , 2.30 (s, 3H) ppm; MS (ES) 435 (M + H). I-58: 2-(4-Fluoro-3-methoxy-5-methyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-acid methyl ester trifluoroacetate salt pyrimidine-5-carboxylic 1H NMR (300 MHz, DMSO) δ 11.69 (s, 1H), 10.03 (s, 1H), 9.98 - 9.66 (m, 1H), 8. 68 (d, J = 2.8, 1H), 7.44 - 7.01 (m, J = 37.0, 5H), 3.81 (d, J = 2.7, 3H), 3. 55 (s, 3H), 1.96 (s, 3H) ppm; MS (ES) 440 (M + H). I-59: 2-(3-Methoxy-4,5-dimethyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine acid methyl ester trifluoroacetate salt - 5-carboxylic 1H NMR (300 MHz, DMSO) δ 11.68 (s, 1H), 10.04 (s, 1H), 9.89 - 9.68 (m, 1H), 8.68 ( s, 1H), 7.44 - 7.17 (m, 3H), 7.13 (s, 1H), 7.06 - 6.86 (m, 1H), 3.82 (s, 3H) ), 3.50 (s, 3H), 2.48 (s, 3H), 1.95 (s, 3H) ppm; MS (ES) 436 (M + H). I-60: 4-(5-Nitro-4-(2-oxo-2,3-dihydrobenzo[d]-oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide trifluoroacetate salt I-61: Salt of 5-(5-fluoro-2-(2-fluoro-3-(2-methoxyethoxy)-4-methylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate I-62: 5-(2-(2-fluoro-3-(2-methoxyethoxy)-4-methylphenylamino)pyrimidin-4-yl-amino)benzo[d]oxazol-2(3H)-one I-63: 5-[5 -Hydroxymethyl-2-(3,4,5-trimethyl-phenylamino)-pyrimidin-4-ylamino]-3H-benzooxazol-2-one 1H NMR (300 MHz, DMSO) δ 11.75 - 11.40 (m, 1H), 8.87 (s, 1H), 8.34 (s, 1H), 7.92 (s, 1H), 7.25 (dd, J = 8.3, 15.4, 3H), 5.12 (t, J = 5.6, 1H), 4.43 (d, J = 5.3, 2H), 2.48 (d, J = 1.7, 6H), 2 0.01 (s, 3H) ppm; MS (ES) 392 (M + H). I-64: 2,6-Dimethyl-4-[5-methyl-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidin-2-ylamino]-benzonitrile trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.75 (s, 1H), 10.47 - 9.99 (m, 1H), 9.55 - 9.23 (m, 1H), 7.96 ( s, 1H), 7.33 (s, 2H), 7.29 - 7.07 (m, 1H), 2.15 (s, 6H), 2.13 (s, 3H) ppm; MS (ES) 387 (M + H). I-65: 5-[5-Methyl-2-(3-vinyl-phenylamino)-pyrimidin-4-ylamino]-3H-benzooxazol-2-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11. 76 (s, 1H), 10.45 (s, 1H), 9.74 (s, 1H), 7.92 (s, 1H), 7.50 (s, 1H), 7. 40 - 7.04 (m, 5H), 6.37 (dd, J = 10.9, 17.6, 1H), 5.50 (d, J = 17.7, 1H), 5.09 (d, J = 10.9, 1H), 2.13 (s, 3H) ppm; MS (ES) 360 (M + H). I-66: 5-(5-Chloro-2-(2-fluoro-3-(2-methoxyethoxy)-4-methylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) trifluoroacetate salt - one I-67: 5-(5-chloro-2-(2-fluoro-3-methoxy-4-methylphenylamino)pyrimidin-4-yl-amino)benzo[d]oxazol-2(3H)-a I- 68: 5-[2-(4-Ethyl-2-fluoro-3-methoxy-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one trifluoroacetate salt 1H NMR (300 MHz , DMSO) δ 11.73 (s, 1H), 10.11 - 9.84 (m, 1H), 9.70 (s, 1H), 7.94 - 7.79 (m, 1H) ), 7.34 - 7.05 (m, 3H), 6.92 (d, J = 8.6, 1H), 3.72 (s, 3H), 2.55 (dd, J = 6. 6, 14.2, 2H), 2.12 (s, 3H), 1.10 (t, J = 7.5, 2H) ppm; MS (ES) 410 (M + H). I-69: 4-(5-Chloro-4-(2-oxo-2,3-dihydrobenzo[d]-oxazol-5-ylamino)pyrimidin-2-ylamino)-2,6-dimethylbenzonitrile] trifluoroacetate salt I-70: 4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-2,6-dimethylbenzonitrile I-71: Salt of 5-(5-chloro-2-(4-ethyl-2-fluoro-3-methoxyphenyl-amino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate I-72: Salt of 2-Fluoro-3-(5-methyl-4-(2-oxo-2,3-dihydro-benzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzonitrile trifluoroacetate I-73: Trifluoroacetate salt of 3-(5-chloro-4-(2-oxo-2,3-dihydrobenzo[d]-oxazol-5-ylamino)pyrimidin-2-ylamino)-2-fluorobenzonitrile I-74: 2-Trifluoroacetate salt of 2- Methyl (4-fluoro-3,5-dimethylphenylamino)-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidine-5-carboxylate I-75: 5-Trifluoroacetate salt (2-(2-fluoro-4-methoxyphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-76: 5-(5-chloro-trifluoroacetate salt) 2-(2-fluoro-4-methoxyphenylamino)-pyrimidin-4-yl amino)benzo[d]oxazol-2(3H)-one I-77: 5-(2-(2-fluoro-3,4-bis(2-methoxyethoxy)phenyl-amino)-5-methylpyrimidine trifluoroacetate salt -4-ylamino)benzo[d]oxazol-2(3H)-one I-78: 5-(5-Chloro-2-(2-fluoro-3,4-bis(2-methoxy-ethoxy) trifluoroacetate salt )phenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-79: 5-(2-(4-fluoro-3,5-dimethylphenylamino)-5-(hydroxymethyl) trifluoroacetate salt )pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-80: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-(hydroxymethyl) trifluoroacetate salt )pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-81: Trifluoroacetate salt 5-(5-(hydroxymethyl)-2-(3-methoxy-4,5-dimethylphenyl-amino) pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-82: 5-(2-(3-(dimethylamino)-4,5-dimethylphenyl-amino)-5-methylpyrimidine trifluoroacetate salt -4-ylamino)benzo[d]oxazol-2(3H)-one I-83: 5-(5-Chloro-2-(3-(dimethylamino)-4,5-dimethyl-phenylamino)pyrimidine trifluoroacetate salt -4-ylamino)benzo[d]oxazol-2(3H)-one I-84: formate salt of 5-(2-(3-(diethylamino)-4,5-dimethylphenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-85: 5-(formate salt) 2-(3-(ethylamino)-4,5-dimethylphenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-86: 5-(5-chloro formate salt) -2-(3-(diethylamino)-4,5-dimethylphenyl-amino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-87: 5-(5-chloro formate salt) -2-(3-(ethylamino)-4,5-dimethylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-88: 5-(2-(3,4,5) -trimethylphenylamino)-5-methylpyrimidin-4-ylamino)-7-(benzo-[d][1,3]dioxol-6-yl)benzo[d]oxazol-2(3H)-one C28H25N5O4. MS (ESI) m/z 496.27 (M+1)+. I-89: 5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-((dimethylamino)methyl)benzo[d]oxazol-2(ditrifluoroacetate salt) 3H)-one C24H28N6O2. MS (ESI) m/z 433.19 (M+1)+. I-90: 7-((diethylamino)methyl)-5-(2-(3,4,5-trimethyl-phenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(ditrifluoroacetate salt) 3H)-one C26H32N6O2. MS (ESI) m/z 461.24 (M+1)+. I-91: 5-(2-(3,4,5-Trimethylphenylamino)-5-methylpyrimidin-4-ylamino)-7-((pyrrolidin-1-yl)methyl)benzo[d]oxazol-2(3H) -one C26H30N6O2. MS (ESI) m/z 459.23 (M+1)+. I-92: 5-(2-(3,4,5-Trimethylphenylamino)-5-methylpyrimidin-4-ylamino)-7-((piperidin-1-yl)methyl)benzo[d]oxazol-2(3H) -one C27H32N6O2. MS (ESI) m/z 473.22 (M+1)+. 1H NMR (300 MHz, DMSO) δ 8.70 (s, 1H, NH), 8.41 (s, 1H, NH), 7.84 (s, 1H, ArH), 7.31 (s , 1H, ArH), 7.21 (s, 3H, ArH), 3.32 (s, 2H, CH2), 2.52 (m, 4H, 2CH2), 2.06 (s, 3H, CH3) , 1.96 (s, 9H, 3CH3), 1.53 (m, 4H, 3CH2). I-93: 5-(2-(3,4,5-Trimethylphenylamino)-5-methylpyrimidin-4-ylamino)-7-((4-methylpiperazin-1-yl)methyl)benzo[d]oxazol-2( 3H)-one C27H33N7O2. MS (ESI) m/z 488.25 (M+1)+. 1H NMR (300 MHz, DMSO) δ 8.67 (s, 1H, NH), 8.34 (s, 1H, NH), 7.81 (s, 1H, ArH), 7.27 (s , 1H, ArH), 7.21 (s, 2H, ArH), 7.10 (s, 1H, ArH), 3.53 (s, 2H, CH2), 2.43 (m, 8H, 4CH2 ), 2.29 (s, 3H, CH3), 2.05 (s, 3H, CH3), 1.95 (s, 9H, 3CH3). I-94: 4-((5-(2-(3,4,5-Trimethylphenylamino)-5-methylpyrimidin-4-ylamino)-2,3-dihydro-2-oxobenzo[d]oxazol-7-yl) tert-butyl methyl)piperazine-1-carboxylate C31H39N7O4. MS (ESI) m/z 574.36 (M+1)+. I-95: 5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-((piperazin-1-yl)methyl)benzo[d] ditrifluoroacetate salt oxazol-2(3H)-one C26H31N7O2. MS (ESI) m/z 474.45 (M+1)+. I-96: 5-(2-(3,4,5-Trimethylphenylamino)-5-methylpyrimidin-4-ylamino)-7-((E)-3-chloroprop-1-enyl)benzo[d]oxazol-2 (3H)-one C24H24ClN5O2. MS (ESI) m/z 450.17 (M+1)+. I-97: 5-(5-((diethylamino)methyl)-2-(3,4,5-trimethylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one ditrifluoroacetate salt C25H30N6O2 . MS (ESI) m/z 447.41 (M+1)+. I-98: 5-(2-(3,4,5-Trimethylphenylamino)-5-((pyrrolidin-1-yl)methyl)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one C25H28N6O2 . MS (ESI) m/z 445.42 (M+1)+. 1H NMR (300 MHz, DMSO) δ 9.88 (s, 1H, NH), 8.89 (s, 1H, NH), 7.85 (s, 1H, ArH), 7.30 (m , 3H, ArH), 7.16 (m, 2H, ArH), 3.55 (s, 2H, CH2), 2.52 (m, 4H, 2CH2), 2.07 (s, 6H, 2CH3), 2.01 (s, 3H, CH3), 1.77 (m, 4H, 2CH2). I-99: 5-(2-(3,4,5-Trimethylphenylamino)-5-((piperidin-1-yl)methyl)pyrimidin-4-yl-amino)benzo[d]oxazol-2(3H)- one C26H30N6O2. MS (ESI) m/z 459.44 (M+1)+. 1H NMR (300 MHz, DMSO) δ 8.93 (s, 1H, NH), 7.85 (s, 1H, NH), 7.30-7.20 (m, 6H, ArH), 3. 32 (s, 2H, CH2), 2.52 (m, 4H, 2CH2), 2.07 (s, 6H, 2CH3), 2.01 (s, 3H, CH3), 1.57 (m, 4H, 2CH2), 1.46 (m, 4H, 2CH2). I-100: 5-(2-(3,4,5-Trimethylphenylamino)-5-((4-methylpiperazin-1-yl)methyl)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) -one C26H31N7O2. MS (ESI) m/z 474.48 (M+1)+. I-101: 5-(2-(3,4-diethoxy-2-fluorophenylamino)-5-methylpyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one C22H22FN5O4. MS (ESI) m/z 440.23 (M+1)+. 1H NMR (300 MHz, DMSO) δ 11.54 (s, 1H, NH), 7.85 (m, br, 2H, 2NH), 7.77 (s, 1H, ArH), 7.32 ( m, 1H, ArH), 7.21 (m, 2H, ArH), 7.09 (d, J = 10.0, 1H, ArH), 6.74 (d, J = 10.0, 1H, ArH ), 4.03 (q, J = 6.7, 2H, CH2), 3.94 (q, J = 6.7, 2H, CH2), 2.06 (s, 3H, CH3), 1.32 (t, J = 6.7, 3H, CH3), 1.19 (t, J = 6.7, 3H, CH3). I-102: 5-(2-(2-fluoro-3,4-d6-dimethoxyphenylamino)-5-methylpyrimidin-4-yl-amino)benzo[d]oxazol-2(3H)-one I-103: Salt of 5-(2-(3-Ethoxy-4,5-dimethylphenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one ditrifluoroacetate I-104: 5-Ditrifluoroacetate salt (5-chloro-2-(3-ethoxy-4,5-dimethylphenyl-amino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-105: 5-(2-(4- fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-yl-amino)benzo[d]oxazol-2(3H)-one I-106: 5-(2-(4-) Methanesulfonic acid salt fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one 1H NMR (300 MHz, DMSO) δ 12.03 (br s, 1H) , 11.80 (s, 1H), 10.05 (s, 1H), 9.78 (s, 1H), 7.87 (s, 1H), 7.36 - 7.25 (m , 3H), 6.97 (br d, J = 7.2 Hz, 1H), 6.92 (br d, J = 3.9 Hz, 1H), 3.65 (s, 3H), 2 .36 (s, 3H), 2.20 (s, 3H), 2.06 (s, 3H); LRMS (M+) m/z 396.14. I-107: Benzenesulfonic acid salt of 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one 1H NMR (300 MHz, DMSO) δ 11.99 (br s, 1H), 11.80 (s, 1H), 10.05 (s, 1H), 9.81 (s, 1H), 7. 87 (s, 1H), 7.65 - 7.62 (m, 2H), 7.39 - 7.33 (m, 4H), 7.29 - 7.26 (m, 2H), 6.96 (br d, J = 7.4 Hz, 1 H), 6.91 (br d, J = 5.6 Hz, 1 H), 3.66 (s, 3H), 2.20 (s, 3H) , 2.06 (s, 3H); LRMS (M+) m/z 396.15. I-108: 5-(2-(3-Methoxy-4,5-dimethylphenyl-amino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one benzenesulfonic acid salt 1H NMR (300 MHz, DMSO) δ 11.91 (s, 1H), 11.80 (s, 1H), 10.04 (s, 1H), 9.83 (s, 1H), 7.85 (s, 1H), 7.66 (br d, J = 3.3, 1H), 7.63 (br d, J = 1.5, 1H), 7.40 - 7.33 (m , 4H), 7.29 - 7.28 (m, 2H), 6.88 (s, 1H), 6.77 (s, 1H), 3.60 (s, 3H), 2.19 ( s, 3H), 2.04 (br s, 6H); LRMS (M+) m/z 392.26. I-109: 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-5-methylpyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 12.06 (br s, 1H), 11.73 (s, 1H), 9.99 (s, 1H), 9.58 (s, 1H), 7. 85 (s, 1H), 7.16 (s, 1H), 7.11 (s, 1H), 6.90 (s, 1H), 6.83 (s, 1H), 3. 58 (s, 3H), 2.28 (s, 3H), 2.18 (s, 3H), 2.04 (br s, 6H); LRMS (M+) m/z 410.09. I-110: 5-(5-fluoro-2-(3-methoxy-4,5-dimethylphenyl-amino)pyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)- trifluoroacetate salt One I-111: Trifluoroacetate Salt 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-5-methylpyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one 1H NMR (300 MHz, DMSO) δ 12.15 (s, 1H), 10.13 (s, 1H), 9.62 (s, 1H), 7.89 (s, 1H), 7.52 (d, J = 12.4Hz, 1H), 7.19 (s, 1H), 6.92 (s, 1H), 6.83 (s, 1H), 3.66 (s, 3H), 2.19 (s, 3H), 2.12 (s, 3H), 2.06 (s, 3H); LRMS (M+) m/z 414.05. I-112: 7-Fluoro-5-(5-fluoro-2-(3-methoxy-4,5-dimethylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 12.01 (s, 1H), 9.73 (s, 1H), 9.38 (s, 1H), 8.20 (d, J = 4.0 Hz, 1H), 7.78 (d, J = 13.0Hz, 1H), 7.26 (s, 1H), 7.16 (s, 1H), 7.06 (s, 1 H), 3.68 (s, 3H), 2.14 (s, 3H), 2.04 (s, 3H); LRMS (M+) m/z 414.05. I-113: 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one 1H NMR (300 MHz, DMSO) δ 11.62 (s, 1H), 8.81 (s, 1H), 8.36 (s, 1H), 7.91 (s, 1H), 7.39 - 7.37 (m, 2H), 7.26 - 7.22 (m, 2H), 7.11 (s, 1H), 3.56 (s, 3H), 2.13 (s, 3H), 2.03 (s, 3H), 1.99 (s, 3H); LRMS (M+) m/z 392.09. I-114: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one sulfuric acid salt 1H NMR (300 MHz, DMSO) δ 11.73 (s, 1H), 9.57 (s, 1H), 9.20 (s, 1H), 7.89 (s, 1H), 7.31 (s, 3H), 7.08 (br d, J = 7.5 Hz, 1H), 7.05 (br d, J = 6.0 Hz, 1H), 3.64 (s, 3H) , 2.17 (s, 3H), 2.05 (s, 3H); LRMS (M+) m/z 396.06. I-115: (5-(2-(3-methoxy-4,5-dimethylphenylamino)-5-methylpyrimidin-4-ylamino)-2-oxobenzo[d]oxazol-3(2H)-yl)methyl phosphate disodium 1H NMR (300 MHz, D2O) δ 7.61 (s, 1H), 7.45 (dd, J = 8.8, 1.8 Hz, 1H), 7.17 (d, J = 1, 8Hz, 1H), 6.91 (d, J = 8.8Hz, 1H), 6.71 (s, 1H), 6.57 (s, 1H), 5.34 (d, J = 5.8 Hz, 2H), 3.55 (s, 3H), 1.95 (br s, 6H), 1.93 (s, 3H); LRMS (M-) m/z 500.22. I-116: 5-(2-(4-fluoro-3-hydroxy-5-methylphenyl-amino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.80 (br s, 1 H), 10.27 (br s, 1 H), 9.85 (br s, 1 H), 9.76 (br s, 1 H ), 7.89 (br s, 1H), 7.35 (d, J = 8.8Hz, 1H), 7.31 - 7.27 (m, 2H), 6.84 (d, J =2.9Hz, 1H), 6.75 (d, J =7.4Hz, 1H), 2.18 (s, 3H), 1.98 (s, 3H); LRMS (M+) m/z 382.27. I-117: 5-(2-(3-hydroxy-4,5-dimethylphenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz , DMSO) δ 11.81 (s, 1H), 10.21 - 10.08 (m, 1H), 9.77 (br s, 1H), 9.37 (s, 1H), 7 .86 (s, 1H), 7.36 - 7.28 (m, 3H), 6.85 (s, 1H), 6.56 (s, 1H), 2.18 (s, 3H) , 2.01 (s, 3H), 1.96 (s, 3H); LRMS (M+) m/z 378.06. I-118: Calcium dihydrogen phosphate salt of (5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)-2-oxobenzo[d]oxazol-3( 2H)-yl)methyl I-119: 5-(5-fluoro-2-(2-fluoro-5-methoxyphenylamino)pyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one I-120 : 5-(2-(3-Fluoro-5-methoxy-4-methylphenyl-amino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.72 (s, 1H), 10.15 (s, 1H), 9.62 (s, 1H), 7.88 (s, 1H), 7.28 (d , J = 8.3 Hz, 1 H), 7.20, (s, 1 H), 7.19 (d, J = 9.8 Hz, 1 H), 7.03 (d, J = 12, 0Hz, 1H), 6.71 (s, 1H), 3.60 (s, 3H), 2.13 (s, 3H), 1.93 (s, 3H); LCMS (m/z): 396 (MH+). I-121: 5-(5-fluoro-2-(3-fluoro-5-methoxy-4-methyl-phenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt I-122: 5-(2-(2-Fluoro-5-methoxy-4-methyl-phenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.70 (s, 1H), 9.89 (s, 1H), 9.59 (s, 1H), 7.83 (s, 1H), 7.28 - 7.12 (m, 4H), 6.98 (d, J = 6.7Hz, 1H), 3.49 (s, 3H), 2.12 (s, 3H), 2.10 (s , 3H); LCMS (m/z): 396 (MH+). I-123: 5-(5-fluoro-2-(2-fluoro-5-methoxy-4-methylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt I- 124: 5-(2-(4-Chloro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz , DMSO) δ 11.72 (s, 1H), 9.94 (s, 1H), 9.47 (s, 1H), 7.86 (s, 1H), 7.33 - 7. 17 (m, 3H), 7.09 (s, 1H), 6.97 (s, 1H), 3.58 (s, 3H), 2.12 (s, 3H), 2.04 (s , 3H); LCMS (m/z): 412 (MH+). I-125: 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-5-methylpyrimidin-4-ylamino)-3-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 10.10 (s, 1H), 9.78 (s, 1H), 7.84 (s, 1H), 7.44 (s, 1H), 7.34 (d, J = 8.5 Hz, 1 H), 7.24 (d, J = 8.4 Hz, 1 H), 6.80 (s, 1 H), 6.73 (s, 1 H) , 3.48 (s, 3H), 3.13 (s, 3H), 2.14 (s, 3H), 1.97 (s, 6H); LCMS (m/z): 406 (MH+). I-126: 5-(2-(3,4,5-Trimethylphenylamino)-5-methylpyrimidin-4-ylamino)-3-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 9.79 (s, 1H), 9.64 (s, 1H), 7.82 (s, 1H), 7.44 (s, 1H), 7.35 (d , J = 8.4 Hz, 1H), 7.24 (d, J = 8.6Hz, 1H), 6.98 (s, 2H), 3.16 (s, 3H), 2.13 (s, 3H), 2.01 (s, 3H), 2.00 (s, 6H); LCMS (m/z): 390 (MH+). I-127: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methylpyrimidin-4-ylamino)-3-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 10.14 (s, 1H), 9.73 (s, 1H), 7.86 (s, 1H), 7.43 (s, 1H), 7 .34 (d, J = 8.6 Hz, 1 H), 7.24 (d, J = 8.6 Hz, 1 H), 6.93 (d, J = 7.3 Hz, 1 H), 6.85 (d, J = 5.4 Hz, 1H), 3.55 (s, 3H), 3.18 (s, 3H), 2.14 (s, 3H), 1.99 (s, 3H); LCMS (m/z): 410 (MH+). I-128: Ethyl 4-(4-(2,3-dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-methylpyrimidin-2-ylamino)-2-methoxy-6-methylbenzoate 1H NMR ( 300 MHz, DMSO) δ 11.58 (s, 1H), 9.07 (s, 1H), 8.38 (s, 1H), 7.90 (s, 1H), 7.32 - 7.13 (m, 5H), 4.19 (q, J = 7.1 Hz, 2H), 3.49 (s, 3H), 2.08 (s, 3H), 1.95 (s, 3H ), 1.22 (t, J = 7.1 Hz, 3H); LCMS (m/z): 450 (MH+). I-129: 4-(4-(2,3-Dihydro-2-oxobenzo[d]-oxazol-5-ylamino)-5-methylpyrimidin-2-ylamino)-2-methoxy-6-alcohol trifluoroacetate salt methylbenzyl 1H NMR (300 MHz, DMSO) δ 11.79 (s, 1H), 10.10 (s, 1H), 9.68 (s, 1H), 7.90 (s, 1H), 7.39 - 7.24 (m, 3H), 6.93 (s, 1H), 6.83 (s, 1H), 4.46 (s, 2H), 3.57 (s, 3H) , 2.19 (s, 3H), 2.15 (s, 3H); LCMS (m/z): 408 (MH+). I-130: Arginine salt of 5-Methyl-N4-[3-(phosphonooxy)methyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl]-N2-(3,4, 5-trimethyl)phenyl-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 10.10 (br, 1H), 8.88 (s, 1H), 8.24 (s, 1H), 7.83 (s, 1H), 7.52 (s, 2H), 7.38 (br, 2H), 7.28 - 7.19 (m, 4H), 5.41 (d, J = 7 .2 Hz, 2H), 3.04 (q, 2H), 2.12 (s, 6H), 2.09 (s, 3H), 2.01 (s, 3H), 1.64 (m, 2H ), 1.52 (m, 2H); LCMS: purity: 91.62%; MS (m/e): 486.33 (MH+). I-131: Benzenesulfonic acid salt of 5-Methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(3,4,5-trimethyl)phenyl- 2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.76 (s, 1H), 9.80 (s, 1H), 9.75 (s, 1H), 7.79 (s, 1H) 1H), 7.57 (d, J = 3.3Hz, 2H), 7.30 (m, 4H), 7.20 (d, J = 6.6Hz, 2H), 6.96 (s , 2H), 2.13 (s, 3H), 2.03 (s, 9H); LCMS: purity: 95.77%; MS (m/e): 376.39 (MH+). I-132: Tris salt of 5-methyl-N4-[3-(phosphonooxy)methyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl]-N2-(3,4, 5-trimethyl)phenyl-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 10.43 (br, 1H), 9.14 (br, 1H), 8.11 (s, 1H), 7.84 (s, 1H), 7.59 (s, 2H), 7.20 (s, 2H), 5.42 (d, J = 7.8 Hz, 2H), 3.44 (s, 6H), 2.14 (s, 6H), 2.09 (s, 3H), 2.02 (s, 3H); LCMS: purity: 97.81%; MS (m/e): 486.30 (MH+). I-133: N4-{3-[2-(N,N-dimethylamino)ethoxyphosphinyloxymethyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl}-5-methyl-N2-(3 ,4,5-trimethyl)phenyl-2,4-pyrimidine-diamine 1H NMR (300 MHz, DMSO) δ 8.90 (br, 1H), 7.81 (s, 1H), 7.35 (br , 1H), 7.29 (d, J = 6.3 Hz, 3H), 7.14 (s, 1H), 6.97 (s, 1H), 5.51 (d, J = 9 0.0Hz, 2H), 3.93 (m, 2H), 3.15 (m, 2H), 2.69 (s, 6H), 2.14 (s, 6H), 2.11 (s, 3H ), 2.04 (s, 3H); LCMS: purity: 97.97%; MS (m/e): 557.40 (MH+). I-134: N4-{3-bis[2-(N,N-dimethylamino)ethoxyphosphinyloxymethyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl}-5-methyl-N2-( 3,4,5-trimethyl)phenyl-2,4-pyrimidine-diamine 1H NMR (300 MHz, DMSO) δ 10.20 (br, 1H), 8.86 (br, 1H), 8.50 ( br, 1H), 7.82 (s, 1H), 7.38 (s, 2H), 7.28 (s, 2H), 5.51 (d, J = 9.3Hz, 2H), 4.08 (br, 2H), 3.52 (br, 4H), 3.01 (s, 8H), 2.14 (s, 6H), 2.10 (s, 3H), 2.04 (s , 3H); LCMS: purity: 98.27%; MS (m/e): 628.63 (MH+). I-135: 5-Methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(3,4,5-trimethyl)phenyl-pamoic acid salt 2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.64 (s, 1H), 9.11 (br, 1H), 8.77 (br, 1H), 8.39 (s, 2H), 8.12 (d, J = 8.4 Hz, 2H), 7.82 (m, 3H), 7.26 (m, 5H), 7.14 (m, 4H), 4.75 ( s, 2H), 2.08 (s, 3H), 2.00 (s, 6H), 1.99 (s, 3H); LCMS: purity: 95.55%; MS (m/e): 376.37 (MH+). I-136: Dipotassium salt of 5-Methyl-N4-[3-(phosphonooxy)methyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl]-N2-(3,4, 5-trimethyl)phenyl-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 7.82 (s, 1H), 7.64 (s, 2H), 7.18 (br, 4H), 5, 39 (d, 2H), 2.14 (s, 6H), 2.08 (s, 3H), 2.02 (s, 3H); LCMS: purity: 99.19%; MS (m/e): 486.38 (MH+). I-137: Methanesulfonic acid salt of 5-Methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(3,4,5-trimethyl)phenyl- 2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.75 (s, 1H), 9.81 (s, 1H), 9.74 (s, 1H), 7.79 (s, 1H), 7.31 (d, J = 9.3Hz, 1H), 7.20 (m, 2H), 6.96 (s, 2H), 2.29 (s, 3H), 2, 13 (s, 3H), 2.02 (s, 9H); LCMS: purity: 99.23%; MS (m/e): 376.24 (MH+). I-138: N2-(3,5-dimethyl-4-methoxycarbonyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.67 (br, 1H), 7.88 (s, 1H), 7.34 (s, 1H), 7.24 (m, 4H), 3.77 (s, 3H), 2.11 (s, 3H), 2.00 (s, 6H); LCMS: purity: 99.74%; MS (m/e): 420.27 (MH+). I-139: N2-ethyl-N2-(3,4,5-trimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2 ,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.80 (s, 1H), 7.53 (s, 1H), 7.42 (s, 1H), 7.28 (s, 2H) ), 6.99 (s, 2H), 3.71 (q, J = 7.2 Hz, 2H), 2.25 (s, 6H), 2.16 (s, 3H), 2.11 (s , 3H), 1.06 (t, J = 6.6 Hz, 3H); LCMS: purity: 93.99%; MS (m/e): 404.33 (MH+). I-140: N2-(4-carboxy-3,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine N2-(4-tert-butoxycarbonyl-3,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 - pyrimidinediamine (500 mg) was suspended in methanol (5 ml). 4.0 M HCl in dioxane (1.0 ml) was added. The solution was heated to 40°C for 2 hours and 50°C for 6 hours. The solution was evaporated and diluted with water (10 ml). NaHCO3 was added to the solution until pH 3. The precipitation was filtered off and washed with water, dried to give the desired acid (440 mg). 1H NMR (300 MHz, DMSO) δ 11.72 (s, 1H), 7.87 (s, 1H), 7.30 (d, J = 8.1Hz, 1H), 7.22 ( s, 2H), 7.15 (s, 2H), 2.12 (s, 3H), 2.05 (s, 6H); LCMS: purity: 99.92%; MS (m/e): 406.32 (MH+). I-141: N2-(4-benzyloxycarbonyl-3,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.73 (s, 1H), 9.86 (br, 1H), 9.50 (br, 1H), 7.87 (s, 1H) , 7.41 - 7.29 (m, 6H), 7.20 (s, 1H), 7.14 (s, 3H), 5.28 (s, 2H), 2.13 (s, 3H) , 1.98 (s, 6H); LCMS: purity: 100%; MS (m/e): 496.33 (MH+). I-142: N2-(3,5-dimethyl-4-hydroxymethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine A N2-(4-carboxy-3,5-dimethyl)-phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 - pyrimidine-diamine (440 mg), BH3 (1.0 M in THF, 4 ml) was added. The mixture was stirred at 0°C to room temperature overnight, then quenched with methanol (10 ml) and 4.0 M HCl in dioxane was added to pH 7. The reaction solution was evaporated and purified by chromatography on column (methanol in dichloromethane = 0 to 30% in 30 minutes) to give the desired benzyl alcohol. 1H NMR (300 MHz, DMSO) δ 8.69 (s, 1H), 8.17 (s, 1H), 7.81 (s, 1H), 7.23 (s, 2H), 7. 14 (m, 2H), 7.06 (t, 1H), 4.34 (s, 2H), 2.11 (s, 6H), 2.06 (s, 3H); LCMS: purity: 88.12%; MS (m/e): 392.20 (MH+). I-143: N2-(3,5-dimethyl-4-methoxymethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine N2-(3,5-dimethyl-4-hydroxy-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 - pyrimidinediamine was dissolved in methanol and treated with 1.0 M HCl in dioxane (0.1 ml). The solution was evaporated and purified by column chromatography (methanol in dichloromethane = 0 to 30% in 30 minutes) to give the benzyl methyl ether. 1H NMR (300 MHz, DMSO) δ 8.82 (s, 1H), 8.28 (s, 1H), 7.84 (s, 1H), 7.26 (s, 2H), 7. 23 (m, 2H), 7.16 (m, 1H), 4.26 (s, 2H), 3.22 (s, 3H), 2.07 (s, 9H); LCMS: purity: 85.20%; MS (m/e): 406.25 (MH+). I-144: 2,3-Dimethyl-benzoic acid N2-(3,4-dimethyl-5-methoxycarbonyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol- 5-yl)-2,4-pyrimidinediamine (1 g) and K 2 CO 3 (1.1 g, 1.2 eq.) were suspended in DMF (10 ml). To the reaction mixture, iodomethane (0.5 ml, 1.2 eq.) was added. The reaction was stirred at room temperature overnight, then diluted with water (80 ml). The solution was extracted with ethyl acetate (80 ml) and evaporated to give the methyl ester. 1H NMR (300 MHz, DMSO) δ 7.49 (d, J = 7.8 Hz, 1 H), 7.33 (d, J = 7.5 Hz, 1 H), 7.15 (t, J = 7.8 Hz, 1H), 3.79 (s, 3H), 2.33 (s, 3H), 2.26 (s, 3H).
[000387] Methyl 2,3-dimethylbenzoate was dissolved in concentrated sulfonic acid (10 ml). KNO3 (808 mg, 1.2 eq.) was added to the solution at 0°C, then warmed slowly to room temperature overnight. The reaction was quenched with water (80 ml), extracted with ethyl acetate (2 x 80 ml). The organic layers were evaporated to give the mixture of ester and nitrated acids (1:1). The mixture was redissolved in DMF (10 ml). K2CO3 (1.1 g) and iodomethane (0.5 ml) were added to the solution. The reaction was stirred at room temperature for three days, then diluted with water (80 ml). It was extracted with ethyl acetate (3 x 80 ml) and evaporated. The residue was purified by column chromatography (EtOAc in hexanes = 0 to 30% in 45 minutes) to give desired nitrobenzoate ester. 1H NMR (300 MHz, DMSO) δ 8.31 (s, 1H), 8.24 (s, 1H), 3.86 (s, 3H), 2.46 (s, 3H), 2.40 (s, 3H).
[000388] Methyl 2,3-dimethyl-5-nitrobenzoate was dissolved in methanol and charged with 10% Pd-C. The mixture was reacted under hydrogen at 40 psi (276 kPa) for one hour. The catalyst was filtered off on celite, washed with methanol and evaporated to give aniline (500 mg). 1H NMR (300 MHz, DMSO) δ 6.74 (d, J = 2.1 Hz, 1H), 6.55 (d, J = 2.4Hz, 1H), 4.99 (s, 2H ), 3.74 (s, 3H), 2.15 (s, 3H), 2.11 (s, 3H).
[000389] 2-Chloro-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-4-pyrimidineamine (500 mg) and 5-amino-2,3 methyl -dimethylbenzoate (500 mg) were suspended in isopropanol (5 ml) and TFA (20 drops). The solution was heated at 100°C overnight in a sealed tube, then cooled to room temperature. LCMS showed complete product conversion. The reaction solution was diluted with 2.0 M NH3 in methanol (10 ml). The solution was sonicated. The precipitation was filtered off and washed with methanol (50 ml) until the filtered solution turned colorless, dried to give the desired product (630 mg). 1H NMR (300 MHz, DMSO) δ 11.55 (s, 1H), 9.07 (s, 1H), 8.40 (s, 1H), 7.86 (s, 1H), 7 .72 (s, 1H), 7.65 (s, 1H), 7.31 (d, J = 8.1 Hz, 2H), 7.17 (d, J = 8.4 Hz, 1H ), 3.67 (s, 3H), 2.20 (s, 3H), 2.08 (s, 3H), 2.04 (s, 3H); LCMS: purity: 97.93%; MS (m/e): 420.38 (MH+). I-145: N2-(3-carboxy-4,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine N2-(3,4-dimethyl-5-methoxycarbonyl)-phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4- pyrimidine-diamine (500 mg) was suspended in THF (6 ml). To the reaction mixture, 1.0 M aqueous solution (6 ml) was added. The reaction was heated at 60°C for two hours, then diluted with methanol (10 ml) and neutralized with 1 M aqueous solution of HCl until pH around 6. The reaction mixture was evaporated and then diluted with water (20 ml). The precipitate was collected by filtration and washed with water, dried to give the desired acid (420 mg). 1H NMR (300 MHz, DMSO) δ 11.58 (s, 1H), 9.14 (s, 1H), 8.53 (s, 1H), 7.86 (s, 1H), 7 .66 (s, 1H), 7.63 (s, 1H), 7.33 (d, J = 8.7Hz, 1H), 7.28 (s, 1H), 7.19 ( d, J = 8.7Hz, 1H), 2.24 (s, 3H), 2.08 (s, 3H), 2.00 (s, 3H); LCMS: purity: 96.84%; MS (m/e): 406.23 (MH+). I-146: N2-(3,5-dimethyl-4-fluoro)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl) benzenesulfonic acid salt )-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.75 (s, 1H), 9.87 (s, 1H), 9.75 (s, 1H), 7.81 ( s, 1H), 7.57 (dd, J = 2.4, 7.2 Hz, 2H), 7.33 - 7.27 (m, 4H), 7.19 (d, J = 6.0 Hz, 2H), 7.04 (d, J = 6.3 Hz, 2H), 2.13 (s, 3H), 2.02 (s, 6H); LCMS: purity: 97.04%; MS (m/e): 380.24 (MH+). I-147: N2-(3,4-dimethyl-5-hydroxymethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine N2-(3-carboxy-4,5-dimethyl)-phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4- pyrimidine-diamine (270 mg) was suspended in anhydrous THF (2 ml), then BH3 (1.0 M in THF, 2 ml) was added to the reaction. The mixture was stirred at room temperature for 3 days, then quenched with methanol (10 ml) and HCl in dioxane (1.0 M, 3 drops). The reaction solution was evaporated and purified by column chromatography (methanol in dichloromethane = 0 to 30% in 30 minutes) to give the desired benzyl alcohol (150 mg). 1H NMR (300 MHz, DMSO) δ 11.56 (s, 1H), 8.77 (s, 1H), 8.26 (s, 1H), 7.84 (s, 1H), 7 .44 (s, 1H), 7.36 (d, J = 9.0Hz, 1H), 7.33 (s, 1H), 7.29 (s, 1H), 7.20 ( d, J = 9.0 Hz, 1H), 4.90 (t, 1H), 4.32 (d, J = 5.4Hz, 2H), 2.06 (s, 3H), 2, 02 (s, 3H), 1.98 (s, 3H); LCMS: purity: 88.60%; MS (m/e): 392.24 (MH+). I-148: N2-(4-n-butyl-3-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.72 (s, 1H), 9.52 (br, 2H), 7.71 (s, 1H), 7.24 - 7.04 (m, 6H), 2.27 (s, 2H), 2.11 (s, 3H), 1.28 (m, 4H), 0.80 (m, 3H); LCMS: purity: 81.74%; MS (m/e): 390.19 (MH+). I-149: N2-(4-bromo-3,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.59 (s, 1H), 9.02 (s, 1H), 8.39 (s, 1H), 7.87 (s, 1H) , 7.43 (s, 2H), 7.27 (s, 1H), 7.24 (s, 2H), 2.09 (s, 6H), 2.07 (s, 3H); LCMS: purity: 97.80%; MS (m/e): 442.13 (MH+).
[000390] I-150: N2-(4-tert-butoxycarbonyl-3,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl )-2,4-pyrimidinediamine 3,5-Dimethylaniline (3 g) was dissolved in dichloromethane (50 ml). To the reaction solution, trifluoroacetic anhydride (4.8 ml, 1.4 eq.) was added in a water bath. After the reaction was stirred at room temperature for 15 minutes, bromine (1.27 ml, 1 eq) was added slowly in a water bath. The reaction was stirred at room temperature for three hours, then quenched with 10% Na2S2O3 (100 ml). The solution was extracted with dichloromethane (3 x 100 ml). The organic layers were dried over MgSO4. Then it was treated with activated charcoal, filtered and evaporated. The residue was crystallized from dichloromethane and hexanes to give N1-(4-bromo-3,5-dimethylphenyl)-2,2,2-trifluoroacetamide (6.2 g, 84% over two steps). 1H NMR (300 MHz, DMSO) δ 11.23 (br, 1H), 7.46 (s, 2H), 2.34 (s, 6H).
[000391] N1-(4-Bromo-3,5-dimethylphenyl)-2,2,2-trifluoroacetamide (3 g) was dissolved in THF (50 ml). At -78°C, MeLi in ether (1.6 M, 8.9 ml, 1.4 eq.) was added to the solution and stirred for 5 minutes. Then s-BuLi in cyclohexanes (1.4 M, 10 ml, 1.4 eq.) was added and stirred for 5 minutes. Boc anhydride (4 g, 1.8 eq.) was then added at -78°C. The reaction solution was allowed to warm to room temperature and stirred for 3.5 hours. The reaction was quenched with water (100 ml). The solution was extracted with dichloromethane (2 x 100 ml). The organic layers were evaporated and purified by column chromatography (EtOAc in hexanes = 0 to 50% in 45 minutes) to give the desired tert-butyl ester. 1H NMR (300 MHz, DMSO) δ 11.22 (s, 1H), 7.35 (s, 2H), 2.23 (s, 6H), 1.52 (s, 9H); 19F NMR (282 MHz, DMSO) δ - 89.51.
[000392] N1-[4-(tert-butoxy)carbonyl-3,5-dimethylphenyl]-2,2,2-trifluoro-acetamide was dissolved in methanol (50 ml) and aqueous NaOH solution (1.0 N, 50 ml). The solution was heated at 60°C for one hour and at room temperature overnight. It was extracted with ethyl acetate (2 x 100 ml), evaporated, purified by column chromatography (EtOAc in hexanes = 0 to 50% in 45 minutes) to give the desired aniline. 1H NMR (300 MHz, DMSO) δ 6.17 (s, 2H), 5.22 (br, 2H), 2.10 (s, 6H), 1.47 (s, 9H).
[000393] 2-Chloro-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-4-pyrimidineamine (500 mg) and 4-amino-2,6 tert-butyl dimethylbenzoate (500 mg) were suspended in isopropanol (5 ml) and TFA (15 drops). The solution was heated at 100°C overnight in a sealed tube, then cooled to room temperature. LCMS showed complete product conversion. The reaction solution was diluted with 2.0 M NH3 in methanol (20 ml). The solution was sonicated. The precipitation was filtered off and washed with methanol (50 ml) until the filtered solution turned colorless, dried to give the desired product (560 mg). 1H NMR (300 MHz, DMSO) δ 11.58 (br, 1H), 9.03 (s, 1H), 8.38 (s, 1H), 7.87 (s, 1H), 7 .30 (s, 2H), 7.24 (d, J = 5.1 Hz, 3H), 2.08 (s, 3H), 2.01 (s, 6H), 1.49 (s, 9H) ; LCMS: purity: 91.21 %; MS (m/e): 462.28 (MH+). I-151: N2-(3,5-dimethyl-4-fluoro)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl) sulfuric acid salt )-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.68 (s, 1H), 9.41 (br, 1H), 9.09 (br, 1H), 7.82 ( s, 1H), 7.29 - 7.23 (m, 3H), 7.15 (d, J = 6.3 Hz, 2H), 2.10 (s, 3H), 2.00 (s, 6H); LCMS: purity: 98.02%; MS (m/e): 380.26 (MH+). I-152: N2-(3-carboxy-4-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.66 (s, 1H), 9.92 (br, 1H), 9.50 (br, 1H), 7.83 (s, 1H), 7 .78 (s, 1H), 7.54 (d, 1H), 7.23 (s, 2H), 7.19 (s, 1H), 7.12 (d, J = 7.8 Hz , 1H), 2.43 (s, 3H), 2.13 (s, 3H); LCMS: purity: 99.90%; MS (m/e): 392.28 (MH+). I-153: N2-(4-fluoro-3-methoxycarbonyl-5-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2 ,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 9.16 (s, 1H), 8.37 (s, 1H), 7.88 (m, 2H), 7.79 (dd, 1H) ), 7.31 (s, 1 H), 7.26 (d, J = 6.0 Hz, 1 H), 7.19 (d, J = 8.4 Hz, 1 H), 3.72 ( s, 3H), 2.08 (s, 3H), 2.04 (s, 3H); 19F NMR (282 MHz, DMSO) δ - 140.96; LCMS: purity: 97.45%; MS (m/e): 424.28 (MH+). I-154: N2-(4-fluoro-3-hydroxymethyl-5-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2 ,4-pyrimidinediamine 1 H NMR (300 MHz, DMSO) δ 11.76 (s, 1 H), 10.29 (s, 1 H), 9.79 (s, 1 H), 7.86 (s, 1 H), 7.31 (d, J = 9.0 Hz, 1H), 7.19 (m, 3H), 7.13 (d, J = 6.3 Hz, 1H), 4.42 ( s, 2H), 2.13 (s, 3H), 1.94 (s, 3H); 19F NMR (282 MHz, DMSO) δ - 144.80; LCMS: purity: 90.56 %; MS (m/e): 396.31 (MH+). I-155: N2-(3-carboxy-4-fluoro-5-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2 ,4-pyrimidinediamine 1 H NMR (300 MHz, DMSO) δ 11.60 (s, 1 H), 9.46 (br, 1 H), 8.84 (br, 1 H), 7.86 (s, 1 H), 7.72 (t, 2H), 7.28 (d, J = 9.6 Hz, 2H), 7.20 (d, J = 8.4 Hz, 1H), 2.09 (s , 3H), 2.00 (s, 3H); LCMS: purity: 94.88%; MS (m/e): 410.21 (MH+). I-156: 5-Methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(3,4,5-trimethyl)phenyl-hydrochloric acid salt 2,4-pyrimidinediamine 1 H NMR (300 MHz, DMSO) δ 11.76 (s, 1H), 9.93 (s, 1H), 9.75 (s, 1H), 7.81 (s, 1H), 7.31 (d, J = 9.3Hz, 1H), 7.20 (s, 2H), 6.97 (s, 2H), 2.12 (s, 3H), 2, 02 (s, 9H); LCMS: purity: 98.50%; MS (m/e): 376.19 (MH+). I-157: 5-(2-(4-Acetylphenylamino)-5-fluoropyrimidin-4-yl-amino)benzo[d]oxazol-2(3H)-one formate salt I-158: 5-formate salt -(2-(4-(1-(cyclopropylamino)ethyl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one I-159: N-cyclobutyl-formate salt 4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo -[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide I-160: 4-(5-fluoro formate salt -4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-N-propylbenzamide I-161: N-cyclopropyl-4-(5-fluoro formate salt) -4-(2-oxo-2,3-dihydro-benzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide I-162: N-ethyl-4-(5-fluoro-formate) salt 4-(2-oxo-2,3-dihydrobenzo[d]-oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide I-163: 4-(5-fluoro-4-(2-oxo) formate salt -2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-N-isopropylbenzamide I-164: N-cyclobutyl-4-(5-fluoro-4-(2-oxo-2,3) -dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-2-methoxybenzamide Formate salt I-165: Salt of N-cyclopropyl-4-(5-fluoro-4-(2-oxo-2,3-dihydro-benzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-2-(trifluoromethyl)benzamide formate I-166: 4-(5-Fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-N-phenyl-2-( trifluoromethyl)benzamide I-167: 5-(2-(4-methyl-3-(methylsulfonyl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.64 (s, 1H), 9.62 (s, 1H), 9.54 (s, 1H), 8.19 - 8.17 (m, 2H) , 8.00 (dd, J = 8.3, 2.4 Hz, 1 H), 7.54 (dd, J = 8.7, 1.8 Hz, 1 H), 7.42 (s, 1 H), 7.32 - 7.26 (m, 2H), 3.19 (s, 3H), 2.58 (s, 3H); ); LRMS (M+) m/z 429.97. I-168: 5-(2-(4-fluoro-3-(methylsulfonyl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.63 (s, 1H), 9.71 (s, 1H), 9.58 (s, 1H), 8.19 - 8.15 (m, 2H), 8. 11 - 8.05 (m, 1H), 7.49 (d, J = 8.7Hz, 1H), 7.44 - 7.38 (m, 2H), 7.28 (d, J = 8.7 Hz, 1H), 3.32 (s, 3H); ); LRMS (M+) m/z 433.94. I-169: 5-(2-(3-Fluoro-5-morpholinophenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one ditrifluoroacetate salt 1H NMR (300 MHz, DMSO ) δ 11.64 (s, 1 H), 9.57 (s, 1 H), 9.38 (s, 1 H), 8.17 (d, J = 4.2 Hz, 1 H), 7 .41 (d, J = 8.6 Hz, 1 H), 7.38 (s, 1 H), 7.26 (d, J = 8.6 Hz, 1 H), 7.14 (d, J = 11.6 Hz, 1H), 6.95 (s, 1H), 6.35 (d, J = 12.4Hz, 1H), 3.69 - 3.66 (m, 4H), 3.01 - 2.98 (m, 4H); LRMS (M+) m/z 441.03. I-170: 5-(2-(3-Fluoro-5-(4-methylpiperazin-1-yl)-phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H) ditrifluoroacetate salt )-one 1H NMR (300 MHz, DMSO) δ 11.56 (s, 1H), 9.58 (s, 1H), 9.41 (s, 1H), 8.16 (d, J = 3.7Hz, 1H), 8.07 (s, 1H), 8.05 - 8.01 (m, 1H), 7.42 - 7.37 (m, 3H), 7.30 ( s, 2H), 7.23 (d, J = 1.7 Hz, 1H), 2.34 (s, 3H); LRMS (M+) m/z 430.99. I-171: 3-(5-fluoro-4-(7-methyl-2-oxo-2,3-dihydro-benzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzenesulfonamide trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.56 (s, 1 H), 9.58 (s, 1 H), 9.41 (s, 1 H), 8.16 (d, J = 3.7 Hz , 1H), 8.07 (s, 1H), 8.05 - 8.01 (m, 1H), 7.42 - 7.37 (m, 3H), 7.30 (s, 2H) , 7.23 (d, J = 1.7 Hz, 1H), 2.34 (s, 3H); LRMS (M+) m/z 430.99. I-172: 5-(2-(3-(Methylsulfonyl)phenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz , DMSO) δ 11.57 (s, 1H), 9.70 (s, 1H), 9.50 (s, 1H), 8.20 - 8.18 (m, 1H), 8. 15 (s, 1H), 8.10 (br d, J = 7.2Hz, 1H), 7.51 - 7.43 (m, 2H), 7.33 (s, 1H), 7 .22 (s, 1H), 3.16 (s, 3H), 2.34 (s, 3H); LRMS (M+) m/z 429.94. I-173: 5-(2-(4-fluoro-3-(methylsulfonyl)phenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.56 (s, 1H), 9.68 (s, 1H), 9.49 (s, 1H), 8.17 (d, J = 3.8 Hz , 1H), 8.14 - 8.10 (m, 2H), 7.38 (t, J = 9.7 Hz, 1H), 7.30 (s, 1H), 7.24 (s , 1H), 3.32 (s, 3H), 2.34 (s, 3H); LRMS (M+) m/z 447.95. I-174: 3-(5-fluoro-4-(7-fluoro-2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzenesulfonamide trifluoroacetate salt 1H NMR ( 300 MHz, DMSO) δ 11.97 (s, 1H), 9.71 (s, 1H), 9.61 (s, 1H), 8.23 (d, J = 3.6Hz, 1 H), 8.16 (br s, 1H), 8.02 (ddd, J = 7.7, 3.0, 3.0 Hz, 1H), 7.86 (dd, J = 13.1 , 1.8 Hz, 1 H), 7.44 (d, J = 7.8 Hz, 1 H), 7.40 (ddd, J = 7.6, 3.0, 3.0 Hz, 1 H ), 7.33 - 7.28 (m, 3H); LRMS (M+) m/z 434.89. I-175: 5-(5-fluoro-2-(3-d3-methoxy-5-methylphenyl-amino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.67 (s, 1H), 9.65 (s, 1H), 9.34 (s, 1H), 8.17 (d, J = 4.0Hz, 1H), 7.45 (d, J = 8.7Hz, 1H), 7.40 (s, 1H), 7.28 (d, J = 8.7Hz, 1H), 7, 08 (s, 1H), 7.05 (s, 1H), 6.36 (s, 1H), 2.16 (s, 3H); LRMS (M+) m/z 385.07. I-176: 5-(2-(3-(Trifluoromethyl)-5-methoxyphenyl-amino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.55 (s, 1H), 9.59 (s, 1H), 9.47 (s, 1H), 8.19 (d, J = 3.8 Hz, 1H), 7.62 (br s, 2H), 7.24 (s, 1H), 7.20 (s, 1H), 6.75 (s, 1H), 3.73 ( s, 3H), 2.32 (s, 3H); LRMS (M+) m/z 450.06. I-177: 5-(2-(3-Methoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.59 (s, 1H), 9.56 (s, 1H), 9.31 (s, 1H), 8.16 (d, J = 4.0Hz, 1H ), 7.30 (s, 1H), 7.21 (d, J = 1.5Hz, 1H), 7.09 (s, 1H), 7.03 (s, 1H), 6 .36 (s, 1H), 3.64 (s, 3H), 2.31 (s, 3H), 2.15 (s, 3H); LRMS (M+) m/z 396.08. I-178: 5-(2-(4-Methoxy-3-methylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.64 (s, 1H), 9.82 (s, 1H), 9.42 (s, 1H), 8.15 (d, J = 4.5Hz, 1H ), 7.37 - 7.31 (m, 3H), 7.20 (s, 1H), 6.87 (d, J = 8.8Hz, 1H), 3.78 (s, 3H) , 2.29 (s, 3H), 2.06 (s, 3H); LRMS (M+) m/z 396.08. I-179: 5-(2-(3-Methoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 12.00 (s, 1H), 9.73 (s, 1H), 9.42 (s, 1H), 8.21 (d, J = 3.9Hz, 1H ), 7.71 (d, J = 13.0Hz, 1H), 7.25 (s, 1H), 7.09 - 7.08 (m, 2H), 6.40 (s, 1H) ), 3.68 (s, 3H), 2.21 (s, 3H); LRMS (M+) m/z 400.07. I-180: 5-(2-(4-Methoxy-3-methylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.99 (s, 1H), 9.72 (s, 1H), 9.29 (s, 1H), 8.17 (br d, J = 4.1 Hz, 1 H), 7.77 (d, J = 13.0Hz, 1H), 7.38 - 7.36 (m, 2H), 7.24 (br s, 1H), 6.88 (d, J = 9.2 Hz, 1H), 3.78 (s, 3H), 2.11 (s, 3H); LRMS (M+) m/z 400.08. I-181: 5-(2-(4-fluoro-3-methoxy-5-methylphenyl-amino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.69 (s, 1H), 9.68 (s, 1H), 9.37 (s, 1H), 8.18 (d, J = 4.1 Hz , 1 H), 7.47 (dd, J = 8.6, 1.9 Hz, 1 H), 7.40 (s, 1 H), 7.28 (d, J = 8.6 Hz, 1 H), 7.21 - 7.16 (m, 2H), 3.67 (s, 3H), 2.11 (s, 3H); LRMS (M+) m/z 400.01. I-182: 5-(2-(3,4-Dimethoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz , DMSO) δ 11.68 (s, 1H), 9.67 (s, 1H), 9.30 (s, 1H), 8.16 (br d, J = 4.1 Hz, 1H ), 7.49 (dd, J = 8.6, 1.9 Hz, 1 H), 7.40 (s, 1 H), 7.27 (d, J = 8.6 Hz, 1 H), 7.11 (s, 1H), 7.09 (s, 1H), 3.65 (s, 3H), 3.63 (s, 3H), 2.07 (s, 3H); LRMS (M+) m/z 412.04. I-183: 5-(2-(3,4-Dimethoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR ( 300 MHz, DMSO) δ 11.62 (s, 1 H), 9.71 (s, 1 H), 9.39 (s, 1 H), 8.16 (d, J = 4.1 Hz, 1 H), 7.35 (s, 1H), 7.21 (s, 1H), 7.09 (s, 1H), 7.04 (s, 1H), 3.66 (s, 3H ), 3.62 (s, 3H), 2.28 (s, 3H), 2.07 (s, 3H); LRMS (M+) m/z 426.08. I-184: 5-(2-(3,4-Dimethoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.99 (s, 1H), 9.65 (s, 1H), 9.28 (s, 1H), 8.19 (br d, J = 3.8 Hz , 1H), 7.80 (d, J = 12.9Hz, 1H), 7.25 (s, 1H), 7.12 (br s, 2H), 3.70 (s, 3H) , 3.67 (s, 3H), 2.13 (s, 3H); LRMS (M+) m/z 430.04. I-185: 5-(2-(3-Chloro-4,5-dimethoxyphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz , DMSO) δ 11.66 (s, 1H), 9.54 (s, 1H), 9.36 (s, 1H), 8.18 (br d, J = 3.7Hz, 1H ), 7.56 (d, J = 1.9 Hz, 1 H), 7.44 (dd, J = 8.3, 1.2 Hz, 1 H), 7.38 (s, 1 H), 7.28 (d, J = 8.3 Hz, 1H), 7.21 (d, J = 1.9 Hz, 1H), 3.70 (s, 3H), 3.69 (s, 3H ); LRMS (M+) m/z 432.17. I-186: 5-(2-(4-(2-Morpholinoethoxy)-3,5-dimethylphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one 1H ditrifluoroacetate salt NMR (300 MHz, DMSO) δ 11.69 (s, 1H), 10.08 (s, 1H), 9.53 (s, 1H), 9.22 (s, 1H), 8. 15 (d, J = 4.0 Hz, 1 H), 7.48 (dd, J = 8.6, 1.6 Hz, 1 H), 7.37 (d, J = 1.6 Hz, 1 H), 7.31 - 7.27 (m, 2H), 4.07 - 3.57 (m, 10H), 3.32 - 3.21 (m, 2H), 2.15 (s, 3H) ; LRMS (M+) m/z 495.11. I-187: 5-(2-(3,5-Dimethylphenylamino)-5-fluoro-pyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 12.00 (s, 1H), 9.64 (s, 1H), 9.31 (s, 1H), 8.20 (d, J = 3.9Hz, 1H ), 7.75 (d, J = 12.7 Hz, 1H), 7.28 (br s, 2H), 7.24 (br s, 1H), 6.61 (s, 1H), 2.20 (s, 6H); LRMS (M+) m/z 384.03. I-188: 5-(2-(3,4,5-Trimethylphenylamino)-5-fluoro-pyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.99 (s, 1H), 9.61 (s, 1H), 9.20 (s, 1H), 8.18 (d, J = 3.9Hz, 1H), 7.76 (d, J = 13.2 Hz, 1H), 7.28 - 7.23 (m, 3H), 2.17 (s, 6H), 2.09 (s, 3H ); LRMS (M+) m/z 398.06. I-189: 5-(2-(4-fluoro-3,5-dimethylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 12.00 (s, 1H), 9.64 (s, 1H), 9.31 (s, 1H), 8.19 (d, J = 3.8Hz, 1H), 7.73 (d, J = 12.7Hz, 1H), 7.35 (s, 1H), 7.32 (s, 1H), 7.24 (br s, 1H ), 2.15 (br s, 6H); LRMS (M+) m/z 402.03. I-190: 5-(2-(4-fluoro-3-methoxy-5-methylphenyl-amino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H) trifluoroacetate salt -one 1H NMR (300 MHz, DMSO) δ 11.62 (s, 1H), 9.59 (s, 1H), 9.35 (s, 1H), 8.16 (d, J = 4 .1Hz, 1H), 7.31 (s, 1H), 7.26 - 7.20 (m, 2H), 7.12 (dd, J = 5.7, 2.0Hz, 1H ), 3.65 (s, 3H), 2.29 (s, 3H), 2.09 (d, J = 1.6 Hz, 3H); LRMS (M+) m/z 414.05. I-191: Trifluoroacetate salt of 5-(2-(4-fluoro-3-methoxy-5-methylphenyl-amino)-5-fluoropyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H) -one 1H NMR (300 MHz, DMSO) δ 12.01 (s, 1H), 9.73 (s, 1H), 9.42 (s, 1H), 8.21 (d, J = 3 0.9 Hz, 1H), 7.75 (dd, J = 13.0, 1.5 Hz, 1H), 7.26 (s, 1H), 7.22 (br d, J = 7. 6Hz, 1H), 7.18 (br d, J = 5.7Hz, 1H), 3.73 (s, 3H), 2.16 (d, J = 1.7Hz, 3H); LRMS (M+) m/z 418.02. I-192: 5-(2-(2,4-Difluoro-3-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.57 (s, 1H), 9.57 (s, 1H), 9.02 (s, 1H), 8.13 (d, J = 4.1Hz, 1H), 7.39 - 7.31 (m, 2H), 7.16 - 7.09 (m, 2H), 3.90 (br s, 3H), 2.22 (s, 3H); LRMS (M+) m/z 418.08. I-193: 5-(2-(2,4-Difluoro-3-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.94 (s, 1 H), 9.66 (s, 1 H), 9.00 (s, 1 H), 8.15 (t, J = 4.1 Hz, 1H), 7.70 (d, J = 13.2 Hz, 1H), 7.33 (br dd, J = 14.3, 8.7 Hz, 1H), 7.19 - 7.09 (m, 2H), 3.92 (s, 3H); 422.04. I-194: 5-(2-(3,5-Dimethoxy-4-methylphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz , DMSO) δ 11.68 (s, 1H), 9.65 (s, 1H), 9.36 (s, 1H), 8.17 (d, J = 4.0Hz, 1H) , 7.52 (dd, J = 8.7, 1.8 Hz, 1 H), 7.43 (d, J = 1.8 Hz, 1 H), 7.25 (d, J = 8. 7Hz, 1H), 7.01 (br s, 2H), 3.61 (s, 6H), 1.95 (s, 3H); LRMS (M+) m/z 412.40. I-195: 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz , DMSO) δ 11.70 (s, 1H), 9.71 (s, 1H), 9.35 (s, 1H), 8.17 (d, J = 4.1Hz, 1H) , 7.49 (dd, J = 8.6, 1.8 Hz, 1 H), 7.41 (d, J = 1.8 Hz, 1 H), 7.27 (d, J = 8.6 Hz, 1H), 7.15 (s, 1H), 7.04 (s, 1H), 3.61 (s, 3H), 2.09 (s, 3H), 2.02 (s, 3H); LRMS (M+) m/z 396.21. I-196: 5-(2-(4-Methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt 1H NMR (300 MHz, DMSO) δ 11, 61 (s, 1H), 9.30 (s, 1H), 8.99 (s, 1H), 8.03 (s, 1H), 7.53 - 7.41 (m, 3H) , 7.36 (s, 1H), 7.21 (d, J = 8.6 Hz, 1H), 6.77 (d, J = 8.9 Hz, 2H), 3.67 (s, 3H); LCMS (m/z): 368 (MH+). I-197: 5-(2-(3-(Difluoromethyl)-4-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt 1H NMR (300 MHz , DMSO) δ 9.31 (s, 1H), 9.16 (s, 1H), 8.07 (s, 1H), 7.74 (d, J = 10.5Hz, 2H), 7 .46 (d, J = 8.7 Hz, 1 H), 7.36 (s, 1 H), 7.19 (d, J = 8.7 Hz, 1 H), 6.98 (t, J = 61.4, 49.4 Hz, 1H), 6.97 (s, 1H), 3.77 (s, 3H); LCMS (m/z): 418 (MH+). I-198: 5-(2-(3-(Difluoromethyl)-5-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz , DMSO) δ 11.58 (s, 1H), 9.44 (s, 2H), 8.13 (s, 1H), 7.44 - 7.39 (m, 3H), 7.34 ( s, 1H), 7.21 (d, J = 8.6Hz, 1H), 6.98 (s, 1H), 6.79 (s, 1H), 6.79 (t, J = 57.0Hz 1H), 6.60 (s, 1H), 3.65 (s, 3H); LCMS (m/z): 418 (MH+). I-199: 5-(2-(3-(Fluoromethyl)-5-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz , DMSO) δ 11.60 (s, 1 H), 9.60 (s, 1 H), 9.45 (s, 1 H), 8.13 (d, J = 3.9 Hz, 1 H) , 7.40 (d, J = 8.7 Hz, 1H), 7.34 - 7.23 (m, 4H), 6.52 (s, 1H), 5.21 (d, J = 47 .7 Hz, 2H), 3.63 (s, 3H); LCMS (m/z): 400 (MH+). I-200: 5-(2-(4-Isopropylphenylamino)-5-fluoro-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.69 (s, 1H), 9.88 (s, 1H), 9.57 (s, 1H), 8.14 (s, 1H), 7.44 - 7.33 (m, 4H), 7.23 (d, J = 8.6 Hz, 1H), 7.09 (d, J = 8.5 Hz, 2H), 2.79 (dt, J = 13.6, 6, 7Hz, 1H), 1.14 (d, J = 6.9Hz, 6H); LCMS (m/z): 380 (MH+). I-201: 5-(2-(4-tert-butylphenylamino)-5-fluoro-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO δ 11.69 (s, 1H), 9.88 (s, 1H), 9.55 (s, 1H), 8.14 (s, 1H), 7.44 - 7.33 ( m, 4H), 7.23 (d, J = 8.6 Hz, 3H), 1.22 (s, 9H); LCMS (m/z): 394 (MH+). I-202: 5-(2-(p-tolylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11, 68 (s, 1H), 9.78 (s, 1H), 9.47 (s, 1H), 8.12 (d, J = 4.3Hz, 1H), 7.46-7 .22 (m, 5H), 7.02 (d, J = 8.1 Hz, 2H), 2.21 (s, 3H); LCMS (m/z): 352 (MH+). I-203: 5-(2-(4-methyl-3-(pyridin-4-yl)phenylamino)-5-fluoropyrimidin-4-yl-amino)benzo[d]oxazol-2(3H)-one 1H NMR (300 MHz, DMSO) δ 11.51 (s, 1H), 9.36 (s, 1H), 9.29 (s, 1H), 8.58 (s, 2H), 8.08 ( s, 1H), 7.64 (s, 1H), 7.56 (d, J = 8.2Hz, 1H), 7.40 - 7.30 (m, 4H), 7.14 ( d, J = 8.5 Hz, 1H), 7.01 (d, J = 8.6 Hz, 1H), 2.14 (s, 3H); LCMS (m/z): 429 (MH+). I-204: 5-(2-(4-methyl-3-(pyridin-3-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one 1H NMR (300 MHz, DMSO) δ 11.52 (s, 1H), 9.42 (s, 1H), 9.33 (s, 1H), 8.58 (s, 1H), 8.52 (s , 1H), 8.09 (s, 1H), 7.84 (d, J = 7.9Hz, 1H), 7.63 (s, 1H), 7.57 - 7.47 ( m, 2H), 7.38 (d, J = 8.7Hz, 1H), 7.29 (s, 1H), 7.15 (d, J = 8.4Hz, 1H), 6 .98 (d, J = 8.6 Hz, 1H), 2.13 (s, 3H); LCMS (m/z): 429 (MH+). I-205: 5-(2-(4-fluoro-3-(pyridin-4-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one 1H NMR (300 MHz, DMSO) δ 11.44 (s, 1H), 9.50 (s, 2H), 8.67 (d, J = 6.2Hz, 2H), 8.13 (s, 1H), 8.03 (s, 1H), 7.69 - 7.61 (m, 3H), 7.34 - 7.22 (m, 3H), 7.04 (d, J = 8.6 Hz, 1 H); LCMS (m/z): 433 (MH+). I-206: 5-(2-(4-fluoro-3-(pyridin-3-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one 1H NMR (300 MHz, DMSO) δ 11.47 (s, 1H), 9.49 (d, J = 5.7Hz, 2H), 8.63 (s, 1H), 8.59 (s, 1H) , 8.13 (s, 1H), 7.92 (d, J = 7.3 Hz, 2H), 7.64 - 7.58 (m, 1H), 7.55 - 7.48 (m , 1H), 7.36 - 7.29 (m, 2H), 7.26 - 7.19 (m, 1H), 6.99 (d, J = 8.7Hz, 1H); LCMS (m/z): 433 (MH+). I-207: 5-(2-(3-Methoxy-4-(pyridin-4-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt 1H NMR (300 MHz, DMSO) δ 9.25 (d, J = 7.4 Hz, 2H), 8.56 (s, 1H), 8.41 (s, 1H), 8.34 (d , J = 7.6Hz, 2H), 7.71 (d, J = 8.7Hz, 1H), 7.36 (s, 1H), 7.16 (s, 2H), 6.73 (s, 2H), 6.38 (d, J = 8.6 Hz, 1H), 6.34 (s, 1H), 3.87 (s, 3H); LCMS (m/z): 445 (MH+). I-208: 5-(2-(4-Methoxy-3-(pyridin-4-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt 1H NMR (300 MHz, DMSO) δ 9.32 (s, 1H), 9.17 (s, 1H), 8.48 (d, J = 5.9 Hz, 2H), 8.25 (s , 1H), 8.06 (d, J = 3.7Hz, 1H), 7.69 (s, 1H), 7.62 (d, J = 8.8Hz, 1H), 7 .41 - 7.34 (m, 4H), 7.02 (d, J = 9.1 Hz, 2H), 3.72 (s, 3H); LCMS (m/z): 445 (MH+). I-209: 5-(2-(4-Methoxy-3-(pyridin-3-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt 1H NMR (300 MHz, DMSO) δ 9.31 (s, 1H), 9.14 (s, 1H), 8.53 (s, 1H), 8.45 (d, J = 4.8 Hz, 1H), 8.06 (d, J = 3.7Hz, 1H), 7.75 (d, J = 8.3Hz, 1H), 7.65 (s, 1H), 7.60 (d, J = 9.0 Hz, 1H), 7.43 - 7.31 (m, 4H), 6.99 (dd, J = 8.7, 5.4 Hz, 2H), 3.71 (s, 3H); LCMS (m/z): 445 (MH+). I-210: 5-fluoro-N2-[3-(1-hydroxy-2,2,2-trifluoroethyl)]phenyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5- il)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.61 (s, 1H), 9.56 (s, 1H), 9.46 (s, 1H), 8.11 (d, J = 3.9, 1H), 7.73 (d, J = 8.7, 1H), 7.62 (s, 1H), 7.47 (dd, J = 2.1 , 8.7, 1H), 7.34 (d, J = 2.4, 1H), 7.22 (d, J = 7.8, 1H), 7.19 (d, J = 7 .8.8, 1H), 7.01 (d, J = 8.1, 1H), 6.77 (br, 1H), 4.93 (q, J = 6.9, 1H); 19F NMR (282 MHz, DMSO) δ - 92.49 (d, J = 9); LCMS: purity: 98.08%; MS (m/e): 436.14 (MH+). I-211: 5-fluoro-N2-(3-methoxy-5-trifluoromethyl)phenyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.53 (br, 1H), 9.52 (s, 1H), 9.44 (s, 1H), 8.14 (d, J = 3.9 , 1H), 7.61 (s, 1H), 7.54 (s, 1H), 7.35 (d, J = 8.7, 1H), 7.32 (s, 1H) , 7.20 (d, J = 8.7, 1H), 6.68 (s, 1H), 3.68 (s, 3H); 19F NMR (282 MHz, DMSO) δ - 76.96, - 179.03; LCMS: purity: 96.31%; MS (m/e): 436.20 (MH+). I-212: 5-fluoro-N2-(4-methoxy-3-trifluoromethyl)phenyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.57 (s, 1H), 9.37 (s, 1H), 9.27 (s, 1H), 8.08 (d, J = 3.6 , 1H), 7.89 (s, 1H), 7.80 (d, J = 9.3, 1H), 7.40 (d, J = 9.0, 1H), 7.33 (s, 1H), 7.19 (d, J = 8.7, 1H), 7.10 (d, J = 8.7, 1H), 3.79 (s, 3H); 19F NMR (282 MHz, DMSO) δ - 76.38, - 180.13; LCMS: purity: 98.02%; MS (m/e): 436.19 (MH+). I-213: N2-[3-(cyclopropylaminocarbonylmethoxy)-4-methoxy]phenyl-5-fluoro-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 - pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.56 (s, 1H), 9.29 (s, 1H), 8.99 (s, 1H), 8.02 (d, J = 3 .6, 1H), 7.88 (d, 1H), 7.47 (d, J = 8.4, 1H), 7.36 (s, 1H), 7.23 (d, J = 8.1, 2H), 7.18 (d, J = 8.7, 1H), 6.83 (d, J = 8.4, 1H), 4.23 (s, 2H), 3 .71 (s, 3H), 2.64 (m, 1H), 0.60 (q, J = 5.7, 2H), 0.45 (m, 2H); 19F NMR (282 MHz, DMSO) δ - 180.69; LCMS: purity: 83.84%; MS (m/e): 481.23 (MH+). I-214: 5-fluoro-N2-(3,4,5-trimethyl)phenyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 9.98 (br, 1H), 9.27 (s, 1H), 8.92 (s, 1H), 8.04 (d, J = 3.6 , 1H), 7.42 (d, J = 8.7, 1H), 7.33 (s, 1H), 7.23 (s, 2H), 7.18 (d, J = 8. 7.1H), 2.05 (s, 6H), 2.00 (s, 3H); 19F NMR (282 MHz, DMSO) δ - 181.06; LCMS: purity: 100%; MS (m/e): 380.13 (MH+). I-215: N2-(3,5-dimethyl-4-fluoro)phenyl-5-fluoro-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.27 (br, 1H), 9.32 (s, 1H), 9.06 (s, 1H), 8.06 (d, J = 3 0.9, 1H), 7.39 (d, J = 8.4, 1H), 7.33 - 7.27 (m, 3H), 7.21 (d, J = 9.0, 1H) ), 2.04(s, 6H); 19F NMR (282 MHz, DMSO) δ -119.33, - 180.66; LCMS: purity: 100%; MS (m/e): 384.10 (MH+). I-216: Lysine salt of 5-Methyl-N4-[3-(phosphonooxy)methyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl]-N2-(3,4, 5-trimethyl)phenyl-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 10.44 (br, 1H), 9.11 (br, 1H), 8.18 (s, 1H), 7.84 (s, 1H), 7.598 (s, 2H), 7.21 (s, 2H), 5.42 (d, J = 7.8 Hz, 2H), 3.10 (t, 1H) ), 2.65 (t, 2H), 2.14 (s, 6H), 2.09 (s, 3H), 2.02 (s, 3H), 1.63-1.56 (m, 2H) , 1.41 (m, 4H); LCMS: purity: 92.76%; MS (m/e): 486.33 (MH+). I-217: N2-(3,4,5-Trimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.39 (br, 1H), 8.70 (s, 1H), 8.29 (s, 1H), 7.83 (s, 1H), 7 .30 (d, J = 6.3, 2H), 7.21 (m, 3H), 2.06 (s, 3H), 1.99 (s, 6H), 1.98 (s, 3H); LCMS: purity: 96.73%; MS (m/e): 376.27 (M + H). I-220: 5-(2-(4-(aminomethyl)phenylamino)-5-methylpyrimidin-4-ylamino)benzo-[d]oxazol-2(3H)-one

[000394] Deuterated compounds were prepared generally as set out above, and further as follows. d4-Thymine (3.00 g, 23.1 mmol) was placed in a pressure vessel and POCl3 (25 ml) was added. The white suspension was heated for 3 hours at 130°C. After approximately 1 hour the suspension became a clear solution. The reaction was then quenched by slowly pouring the reaction mixture into a beaker containing crushed ice. The aqueous phase was then extracted with DCM (3 x 50 ml) and dried over MgSO4. After evaporation of solvents the reaction product was obtained as a white solid (3.72 g, 96%). JAK inhibitors were prepared by the standard sequence of 1st SNAr reaction to LHS aniline carbamate reaction and 2nd SNAr reaction to RHS aniline or d3-aniline (deuterated anilines were prepared in the same way as non-anilines deuterated using CD3I instead of MeI). I-221: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-6-D-5-trideuteromethyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one
MS (ES) 400 (M + H), 398 (M - H). I-222: 5-(2-(4-fluoro-3-trideuteromethoxy-5-methylphenylamino)-6-D-5-tri-deuteromethylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one
1H NMR (300 MHz, DMSO) δ 11.96 (s, 1H), 10.75 (s, 1H), 10.04 (s, 1H), 7.37 - 7.13 (m, 3H ), 6.88 (ddd, J = 7.6, 6.5, 2.0 Hz, 2H), 1.95 (s, 3H); MS (ES) 403 (M + H), 401 (M - H). I-223: 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-6-D-5-trideuteromethyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one
I-224: 5-(2-(3-trideuteromethoxy-4,5-dimethylphenylamino)-6-D-5-trideutero-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one
1H NMR (300 MHz, DMSO) δ 11.78 (s, 1H), 10.22 (s, 1H), 9.80 (s, 1H), 7.24 (m, 3H), 6, 82 (s, 1H), 6.71 (s, 1H), 1.97 (s, 6H); MS (ES) 399 (M + H), 397 (M - H). I-225: 5-(2-(4-(hydroxymethyl)-3-methoxy-5-methylphenylamino)-6-D-5-trideutero-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)- whoa
1H NMR (300 MHz, DMSO) δ 8.82 (s, 1H), 8.31 (s, 1H), 7.33 (m, 2H), 7.26 - 7.02 (m, 3H) , 4.37 (s, 2H), 4.31 (br s, 1H), 3.50 (s, 3H), 2.10 (s, 3H); MS (ES) 412 (M + H), 410 (M - H). I-226: 5-(2-(4-fluoro-3-trideuteromethoxy-5-methylphenyl-amino)-6-D-5-trideuteromethylpyrimidin-4-ylamino)-2-oxobenzo-[di-tert-butyl phosphate d]oxazol-3(2H)-yl)methyl
1H NMR (300 MHz, CDCl 3 ) δ 9.11 (s, 1H), 8.82 (s, 1H), 7.58 (d, J = 7.4Hz, 1H), 7.10 ( d, J = 8.6 Hz, 1 H), 7.05 (d, J = 5.7 Hz, 1 H), 6.75 (d, J = 8.6 Hz, 1 H), 6.43 (s, 1H), 5.78 (d, J = 9.8 Hz, 2H), 2.22 (s, 3H), 1.40 (s, 18H); MS (ES) 625 (M + H), 623 (M - H). I-227: 5-(2-(4-fluoro-3-trideuteromethoxy-5-methylphenylamino)-6-D-5-trideuteromethylpyrimidin-4-ylamino)-2-oxobenzo[d]-oxazole tert-butyl hydrogen phosphate -3(2H)-yl)methyl
I-228: 5-(2-(4-fluoro-3-trideuteromethoxy-5-methylphenyl-amino)-6-D-5-trideuteromethylpyrimidin-4-ylamino)-2-oxobenzo[d]oxazol-3 dihydrogen phosphate (2H)-yl)methyl
MS (ES) 513 (M + H), 511 (M - H). I-229: (5-(2-(4-fluoro-3-trideuteromethoxy-5-methylphenylamino)-6-D-5-trideuteromethylpyrimidin-4-ylamino)-2-oxobenzo[d]oxazol-3(2H)- il) disodium methyl phosphate
1H NMR (300 MHz, D2O) δ 7.33 (d, J = 8.8 Hz, 1 H), 7.07 (s, 1 H), 6.76 (d, J = 8.7 Hz, 1 H), 6.59 (d, J = 5.9 Hz, 1 H), 6.47 (d, J = 7.2 Hz, 1 H), 5.34 (d, J = 5.6 Hz, 2H), 1.82 (s, 3H); MS (ES) 513 (M + H), 511 (M - H). I-230: 4-(4-(2,3-Dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-methyl-pyrimidin-2-ylamino)-2-methoxy-6-methylbenzyl alcohol
1H NMR (300 MHz, DMSO) δ 11.60 (s, 1H), 8.88 (s, 1H), 8.35 (s, 1H), 7.89 (s, 1H), 7 .35 (d, J = 8.2 Hz, 1 H), 7.34 (s, 1 H), 7.22 (d, J = 8.2 Hz, 1 H), 7.17 (s, 1 H), 7.10 (s, 1H), 4.36 (m, 3H), 3.51 (s, 3H), 2.12 (s, 3H), 2.10 (s, 3H). LCMS (m/z): 408.4 (MH+). I-231: 4-(4-(2,3-Dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-methyl-pyrimidin-2-ylamino)-2-methoxy-6-methylbenzoic acid
1H NMR (300 MHz, DMSO) δ 12.71 (br s, 1H), 11.71 (br s, 1H), 9.74 (br s, 1H), 9.26 (br s, 1H ), 7.89 (s, 1H), 7.27 (s, 3H), 7.06 (s, 1H), 6.96 (s, 1H), 3.55 (s, 3H), 2.15 (s, 3H), 2.00 (s, 3H). LCMS (m/z): 422.2 (MH+). II-1: 5-(5-Fluoro-2-(1-oxo-1,2,3,4-tetrahydroiso-quinolin-6-ylamino)pyrimidin-4-ylamino)benzo[d]oxazole trifluoroacetate salt 2(3H)-one II-2: 5-(5-fluoro-2-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7] diformate salt annulen-2-ylamino)pyrimidin-4-ylamino)benzo[d]-oxazol-2(3H)-one II-3: 5-(5-fluoro-2-(7-oxo-6,7) diformate salt ,8,9-tetrahydro-5H-benzo[7]anulen-2-ylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one II-4: Formate salt of (Z)-5 -(5-fluoro-2-(2-methyl-1-oxo-1,2,3,6-tetrahydro-benzo[c]azocin-9-ylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2 (3H)-one II-5: 5-(5-fluoro-2-(5-oxo-6,7,8,9-tetrahydro-5H-benzo-[7]annulen-2-ylamino) formate salt pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one II-6: 5-(2-(Naphthalen-2-ylamino)-5-nitropyrimidin-4-ylamino)benzo[d trifluoroacetate salt ]oxazol-2(3H)-one II-7: 2-(Naphthalen-2-ylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino) acid methyl ester trifluoroacetate salt -pyrimidine-5-carboxylic 1H NMR (30 0 MHz, DMSO) δ 11.67 (s, 1H), 10.29 - 10.14 (m, 1H), 9.99 (s, 1H), 8.75 (s, 1H), 8.22 (s, 1H), 8.01 - 7.83 (m, 1H), 7.76 (s, 2H), 7.68 - 7.56 (m, 1H), 7.41 - 7.24 (m, 4H), 7.25 - 6.99 (m, 1H), 3.85 (s, 3H) ppm; MS (ES) 428 (M + H). II-8: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(1-oxo-1,3-dihydro-isobenzofuran-6-yl )-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.50 (br, 1H), 9.38 (s, 1H), 8.39 (s, 1H), 8.24 ( s, 1H), 7.90 (d, J = 9.6 Hz, 2H), 7.41 (d, J = 8.7 Hz, 1H), 7.30 (t, J = 2.4 Hz, 2H), 7.21 (d, J = 9.3 Hz, 1H), 5.27 (s, 2H), 2.10 (s, 3H); LCMS: purity: 88.61%; MS (m/e): 390.40 (MH+). II-9: N2-(1,3-dihydro-isobenzofuran-5-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2, 4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.42 (br, 1H), 9.05 (s, 1H), 8.35 (s, 1H), 7.86 (s, 1H) ), 7.71 (s, 1H), 7.36 (d, J = 8.4 Hz, 1H), 7.25 (m, 3H), 7.03 (d, J = 8.4 Hz , 1H), 4.87 (s, 2H), 4.76 (s, 2H), 2.07 (s, 3H); LCMS: purity: 97.34%; MS (m/e): 376.30 (MH+). II-10: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(1-oxo-1,3-dihydro-isobenzofuran-5-yl )-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.69 (s, 1H), 10.04 (br, 1H), 8.98 (br, 1H), 7.95 ( s, 2H), 7.59 (m, 2H), 7.36 (d, J = 8.4 Hz, 1H), 7.26 (m, 2H), 5.14 (s, 2H), 2 .13 (s, 3H); LCMS: purity: 81.74%; MS (m/e): 390.19 (MH+). II-11: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(1-oxo-1,3-dihydro-2H-isoindole-6 -yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.34 (br, 1H), 9.19 (s, 1H), 8.46 (s, 1H), 8. 31 (s, 1H), 8.02 (s, 1H), 7.90 (s, 1H), 7.81 (d, J = 7.5Hz, 1H), 7.36 (d , J = 6.6 Hz, 2H), 7.30 (d, J = 8.7 Hz, 1H), 7.22 (d, J = 9.0 Hz, 1H), 4.24 (s , 2H), 2.09 (s, 3H); LCMS: purity: 100%; MS (m/e): 389.21 (MH+). II-12: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(1-oxo-1,3-dihydro-7-methylisobenzofuran-4 -yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.60 (s, 1H), 9.73 (br, 1H), 9.53 (br, 1H), 7. 79 (s, 1 H), 7.70 (d, J = 8.4 Hz, 1 H), 7.46 (d, J = 8.1 Hz, 1 H), 7.14 (m, 3H) , 5.32 (s, 2H), 2.40 (s, 3H), 2.12 (s, 3H); LCMS: purity: 90.64%; MS (m/e): 404.26 (MH+). II-13: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(1-oxo-1,3-dihydro-4-methylisobenzofuran-6 -yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.56 (s, 1H), 9.96 (br, 1H), 9.37 (br, 1H), 7. 90 (s, 1H), 7.77 (s, 1H), 7.58 (s, 1H), 7.27 - 7.19 (m, 3H), 5.28 (s, 2H), 2.13 (s, 3H), 2.12 (s, 3H); LCMS: purity: 97.52%; MS (m/e): 404.29 (MH+). II-14: N2-(4,5-dimethylpyridin-2-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4- pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.71 (s, 1H), 9.27 (s, 1H), 8.84 (s, 2H), 8.31 (s, 1H), 8 .20 (s, 1H), 7.32 (s, 1H), 7.28 (s, 1H), 6.86 (s, 1H), 2.30 (s, 3H), 2. 28 (s, 3H), 2.11 (s, 3H); LCMS: purity: 96.43%; MS (m/e): 363.38 (MH+). II-15: N2-(4,6-dimethylpyridin-2-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4- pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.79 (s, 1H), 9.67 (s, 1H), 8.06 (s, 1H), 7.76 (m, 1H), 7.48 (m, 2H), 7.28 (m, 2H), 6.89 (m, 1H), 2.24 (s, 3H), 2.19 (s, 3H); LCMS: purity: 95.84%; MS (m/e): 363.39 (MH+). II-16: N2-(5-cyano-6-methylpyridin-2-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2, 4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 8.01 (s, 1H), 7.67 (m, 1H), 7.46 (m, 2H), 7.37 (s, 1H) , 2.17 (s, 3H); LCMS: purity: 86.13%; MS (m/e): 374.36 (MH+). II-17: N2-(2-methoxypyridin-4-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.65 (s, 1H), 10.16 (br, 1H), 9.11 (br, 1H), 7.98 (s, 1H), 7, 89 (d, J = 6.0Hz, 1H), 7.31 - 7.10 (m, 5H), 3.72 (s, 3H), 2.14 (s, 3H); LCMS: purity: 97.39%; MS (m/e): 365.35 (MH+). II-18: 5-methyl-N2-(2-methylpyridin-4-yl)-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.67 (s, 1H), 10.75 (s, 1H), 8.79 (s, 1H), 8.28 (d, 1H), 8. 07 (s, 1H), 7.93 (br, 1H), 7.71 (br, 1H), 7.32 (d, J = 8.1 Hz, 1H), 7.27 (s , 1H), 7.25 (d, J = 8.4Hz, 1H), 2.34 (s, 3H), 2.16 (s, 3H); LCMS: purity: 98.95%; MS (m/e): 349.35 (MH+). II-19: N2-(2,6-dimethylpyridin-4-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4- pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.69 (s, 1H), 10.68 (s, 1H), 8.79 (s, 1H), 8.05 (s, 1H), 7.62 (br, 2H), 7.33 (d, J = 8.4 Hz, 1H), 7.24 (d, J = 8.1 Hz, 2H), 2.34 (s, 6H) , 2.15 (s, 3H); LCMS: purity: 99.04%; MS (m/e): 363.37 (MH+). II-20: N2-(6-methoxypyridin-2-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.80 (s, 1H), 8.09 (s, 1H), 7.71 (m, 2H), 7.48 (m, 2H), 7.29 ( s, 1H), 6.60 (d, 1H), 3.92 (s, 3H), 2.18 (s, 3H); LCMS: purity: 90.96%; MS (m/e): 365.34 (MH+). II-21: N2-(5,6-dimethylpyridin-2-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4- pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.80 (s, 1H), 11.14 (br, 1H), 9.71 (s, 1H), 8.07 (s, 1H), 7.63 (d, J = 7.8 Hz, 1H), 7.35 (d, J = 9.0 Hz, 1H), 7.29 (m, 2H), 7.02 (d, J = 8.7Hz, 1H), 2.52 (s, 3H), 2.20 (s, 3H), 2.19 (s, 3H); LCMS: purity: 98.29%; MS (m/e): 363.35 (MH+). II-22: N2-(5-cyano-4-methylpyridin-2-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2, 4-pyrimidinediamine 1H NMR (300 MHz, DMSO) δ 11.73 (s, 1H), 8.63 (s, 1H), 7.99 (s, 1H), 7.30 (s, 2H) , 6.53 (br, 4H), 2.29 (s, 3H), 2.17 (s, 3H); LCMS: purity: 78.01%; MS (m/e): 374.32 (MH+). II-23: 5-(5-Fluoro-2-(1-oxo-2,3-dihydro-1H-benzo[c]-azepin-7-ylamino)pyrimidin-4-ylamino)benzo[d formate salt ]oxazol-2(3H)-one II-24: 5-(5-fluoro-2-(2-methyl-1-oxo-2,3-dihydro-1H-benzo-[c]azepin-) formate salt 7-ylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one II-25: 7-(4-(2,3-dihydro-2-oxobenzo[d]oxazol-formate salt) 5-ylamino)-5-fluoropyrimidin-2-ylamino)-2H-benzo[b][1,4]oxazin-3(4H)-one 1H NMR (300 MHz, DMSO) δ 11.57 (s, 1H) ), 10.49 (s, 1H), 9.46 (s, 1H), 9.22 (s, 1H), 8.07 (s, 1H), 7.39 - 7.33 ( m, 3H), 7.21 (d, J = 8.3 Hz, 1 H), 7.08 (d, J = 8.4 Hz, 1 H), 6.69 (d, J = 8.5 Hz, 1H), 4.44 (s, 2H); LCMS (m/z): 409 (MH+). II-26: 6-(4-(2,3-Dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-fluoropyrimidin-2-ylamino)-3,4-dihydroquinolin-2(trifluoroacetate salt) 1H)-one 1H NMR (300 MHz, DMSO) δ 11.65 (s, 1H), 9.95 (s, 1H), 9.82 (s, 1H), 9.49 (s, 1H) H), 8.12 (s, 1H), 7.40 - 7.33 (m, 3H), 7.23 (d, J = 8.7 Hz, 2H), 6.71 (d, J = 8.5 Hz, 1H), 2.66 (t, J = 7.4 Hz, 2H), 2.36 (t, J = 7.5 Hz, 2H); LCMS (m/z): 407 (MH+). II-27: 7-(4-(2,3-Dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-fluoropyrimidin-2-ylamino)-4,5-dihydro-1H- trifluoroacetate salt benzo[b]azepin-2(3H)-one 1H NMR (300 MHz, DMSO) δ 11.65 (s, 1H), 9.71 (s, 1H), 9.45 (s, 1H) , 9.31 (s, 1H), 8.13 (s, 1H), 7.46 (s, 1H), 7.42 - 7.30 (m, 3H), 7.24 (d, J = 8.6 Hz, 1H), 6.80 (d, J = 8.6Hz, 1H), 2.48 - 2.44 (m, 2H, superimposed with DMSO peak), 2, 08 (t, J = 7.1 Hz, 2H), 1.98 (dd, J = 13.5, 6.7 Hz, 2H); LCMS (m/z): 421 (MH+). II-28: 7-(4-(2,3-Dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-fluoropyrimidin-2-ylamino)-4,5-dihydro-1-trifluoroacetate salt methyl-1H-benzo[b]-azepin-2(3H)-one 1H NMR (300 MHz, DMSO) δ 11.64 (s, 1H), 9.66 (s, 1H), 9.48 ( s, 1H), 8.13 (s, 1H), 7.48 - 7.36 (m, 3H), 7.32 (s, 1H), 7.24 (d, J = 8.6 Hz, 1H), 7.13 (d, J = 8.5Hz, 1H), 3.15 (s, 3H), 2.40 (t, J = 6.5Hz, 2H), 2, 07 (t, J = 6.9 Hz, 2H), 1.93 (dd, J = 13.5, 7.1 Hz, 2H); LCMS (m/z): 435 (MH+). II-29: 5-(5-Methyl-2-(2-morpholinopyridin-4-yl-amino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one ditrifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.70 (s, 1H), 10.41 (s, 1H), 8.80 (s, 1H), 8.10 (s, 1H), 7.85 (d , J = 7.0 Hz, 1H), 7.66 (s, 1H), 7.36 (s, 1H), 7.31 (br s, 2H), 7.19 (d, J = 7.0Hz, 1H), 3.66 - 3.63 (m, 4H), 3.25 - 3.22 (m, 4H), 2.20 (s, 3H); LRMS (M+) m/z 419.98. II-30: 5-(5-Fluoro-2-(2-morpholinopyridin-4-ylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one ditrifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.71 (s, 1H), 10.51 (s, 1H), 9.78 (s, 1H), 8.32 (d, J = 3.5Hz, 1H), 7.89 (d, J = 7.0 Hz, 1H), 7.65 (s, 1H), 7.43 - 7.38 (m, 2H), 7.32 (d, J = 8, 5Hz, 1H), 7.20 (d, J = 7.0Hz, 1H), 3.69 - 3.66 (m, 4H), 3.32 - 3.30 (m, 4H); LRMS (M+) m/z 423.96. II-31: 5-(5-Methyl-2-(pyridin-4-ylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one ditrifluoroacetate salt 1H NMR (300 MHz, DMSO) δ 11.75 (br s, 1H), 10.92 (s, 1H), 8.87 (br s, 1H), 8.46 (d, J = 5.9 Hz, 2H), 8 .16 - 8.10 (m, 2H), 7.38 - 7.36 (m, 2H), 7.32 (d, J = 8.6 Hz, 1H), 6.82 (d, J = 6.3Hz, 1H), 2.22(s, 3H); LRMS (M+) m/z 335.03. Example 6: Xinaphoic Acid Salt
[000395] Preparative Example: The following example illustrates the preparation of a xinafoate salt of a 2,4-pyrimidinediamine described herein. A suspension of a 2,4-pyrimidinediamine as described herein (1 equiv) in a ketone solvent (about 20 ml solvent/g 2,4-pyrimidinediamine, eg MEK), is heated between about 50°C and about 60°C. Water (about 1 ml water/g 2,4-pyrimidinediamine) is added, resulting in a solution. The solution can be passed through a filter for clarification if needed. The solution is held at between about 50°C and about 60°C for between about 30 minutes and about 1.5 hours. A homogeneous solution of 1-hydroxy-2-naphthoic acid (1.1 equiv) in a ketone solvent (about 4 ml solvent/g naphthoic acid, eg MEK) is added which results in precipitation of a solid thereafter. about 5 minutes and 15 minutes. The reaction is cooled to room temperature, stirred for between about 15 h to about 20 hours, and then cooled to between about 0°C and about 10°C for between about 1 h and about 2 hours. The precipitated xinafoic acid salt is then filtered and collected. The filtered solid is washed, for example twice with a ketone solvent (about 2 ml/g, for example MEK) and dried under reduced pressure at between about 40°C and about 60°C, for between about 10 and about 20 hours. The xinafoic acid salt is suitably processed, e.g., micronized and formulated as described herein, e.g., into a formulation for administration by inhalation. In a particular embodiment, compound I-217 is the 2,4-pyrimidinediamine used in this Example. Example 7: Assay for CD23 Expression in IL-4 Stimulated Ramos B Cells
[000396] B cells stimulated with the cytokine Interleukin-4 (IL-4) activate the JAK/Stat pathway through phosphorylation of the family of JAK kinases, JAK-1 and JAK-3, which in turn phosphorylate and activate the Stat-6 transcription factor. One of the genes upregulated by activated Stat-6 is the low-affinity IgE receptor, CD23. To study the effect of inhibitors on the JAK kinase family, human Ramos B cells were stimulated with human IL-4 and CD23 surface expression was measured.
[000397] The Ramos B cell line was purchased from ATCC (ATCC Catalog No CRL-1596). Cells were cultured in RPMI 1640 (Cellgro, MediaTech, Inc., Herndon, VA, Cat No 10-040-CM) with 10% FBS, heat inactivated (JRH Biosciences, Inc, Lenexa, Kansas, Cat No 12106- 500M) according to the ATCC propagation protocol. Cells were maintained at a density of 3.5 x 105. On the day before the experiment, Ramos B cells were diluted to 3.5 x 105 cells/ml to ensure they were in a logarithmic growth phase.
[000398] Cells were spun and suspended in RPMI with 5% serum. 5 x 104 cells were used per dot in a 96-well tissue culture plate. Cells were preincubated with compound or DMSO vehicle control (Sigma-Aldrich, St. Louis, MO, Cat No D2650) for 1 hour in a 37°C incubator. Cells were then stimulated with IL-4 (Peprotech Inc., Rocky Hill, NJ, Cat No 200-04) to a final concentration of 50 units/ml for 20 to 24 hours. Cells were then spun and stained with anti-CD23-PE (BD Pharmingen, San Diego, CA, Cat No 555711) and analyzed by FACS. Detection was performed using a BD LSR I System flow cytometer purchased from Becton Dickinson Biosciences of San Jose, California. The IC50 calculated based on the results of this assay are given in Table IX. Example 8: Assay for IL-2 Stimulated Human Primary T Cell Proliferation
[000399] Primary human T cells derived from peripheral blood and preactivated through stimulation of the T cell receptor and CD28, proliferated in vitro in response to the cytokine Interleukin-2 (IL-2). This proliferative response is dependent on the activation of JAK-1 and JAK-3 tyrosine kinases, which phosphorylates and activates the transcription factor Stat-5.
[000400] Primary human T cells were prepared as follows. Whole blood was obtained from a healthy volunteer, mixed 1:1 with PBS, layered on Ficoll Hypaque (Amersham Pharmacia Biotech, Piscataway, NJ, Catalog #17-1440-03) in a 2:1 ratio of blood/PBS:ficoll and centrifuged for 30 min at 4°C at 1750 rpm. Lymphocytes at the serum:ficoll interface were retrieved and washed twice with 5 volumes of PBS. Cells were resuspended in Yssel's medium (Gemini Bioproducts, Woodland, CA, Catalog #400-103) containing 40 U/ml of recombinant IL2 (R and D Systems, Minneapolis, MN, Catalog #202-ILA (20 μg) )) and seeded in a vial precoated with 1 µg/ml anti-CD3 (BD Pharmingen, San Diego, CA, Catalog #555336) and 5 µg/ml anti-CD28 (Immunotech, Beckman Coulter of Brea California, California, Catalog #IM1376). Primary T cells were stimulated for 3 to 4 days, then transferred to a new flask and kept in RPMI with 10% FBS and 40 U/ml IL-2.
[000401] The day prior to assay setup, primary T cells were centrifuged and resuspended in fresh RPMI with 10% FBS but no IL-2 and starved overnight. For the assay, primary T cells were centrifuged and resuspended in Yssel's medium at 2 x 106 cells/ml. 50 µL of cell suspension containing 80 U/ml IL-2 was added to each well of a 96-well black flat-bottomed plate. For the unstimulated control, IL-2 was omitted from the last column on the plate. Compounds were serially diluted in dimethyl sulfoxide (DMSO, 99.7% pure, cell culture tested, Sigma-Aldrich, St. Louis, MO, Catalog No. D2650) at 5 mM in 3-fold dilutions, and then diluted 1:250 in Yssel's medium. 50 µL of 2X compound was added per well in duplicate and cells were allowed to proliferate for 72 hours at 37°C.
[000402] Proliferation was measured using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega), which determines the number of viable cells in culture based on the quantification of ATP present, as an indicator of metabolically active cells. The substrate was thawed and allowed to come to room temperature. After mixing Cell-Titer-Glo reagent and diluent together, 100 µL was added to each well. Plates were mixed on an orbital shaker for two minutes to induce lysis and incubated at room temperature for another ten minutes to let the signal equilibrate. Detection was performed using a Wallac Victor2 1420 multi-label counter purchased from Perkin Elmer, Shelton, CT. Example 9: Assay for ICAM1 Expression in IFNY Stimulated A549 Epithelial Cells
[000403] Lung epithelial cells, A549, upregulate surface expression of ICAM-1 (CD54) in response to a variety of different stimuli. Therefore, using ICAM-1 expression as readout, the compound's effects on different signaling pathways can be evaluated in the same cell type. IFNy upregulates ICAM-1 through activation of the JAK/Stat pathway. In this example, up-regulation of ICAM-1 by IFNY was evaluated.
[000404] Lung epithelial carcinoma cell line A549 originated from the American Type Culture Collection. Routine culture was with F12K medium (Mediatech Inc., Lenexa, KS, Cat. No. 10-025-CV) with 10% fetal bovine serum, 100 I.U. of penicillin and 100 ng/ml of streptomycin (F12k complete medium). Cells were incubated in a humidified atmosphere of 5% CO2 at 37°C. Prior to use in the assay, A549 cells were washed with PBS and trypsinized (Mediatech Inc., Cat. No. 25-052-CI) to lift the cells. cells. The trypsin cell suspension was neutralized with complete F12K medium and centrifuged to pellet the cells. The cell pellet was resuspended in complete F12K medium at a concentration of 2.0 x 105/ml. Cells were seeded at 20,000 per well, 100 µL total volume, in a flat-bottomed tissue culture dish and allowed to adhere overnight.
[000405] On day two, A549 cells were preincubated with test compound or DMSO (control) (Sigma-Aldrich, St. Louis, MO, Catalog No. D2650) for 1 hour. Cells were then stimulated with IFNY (75 ng/ml) (Peprotech Inc., Rocky Hill, NJ, Cat. No. 300-02) and allowed to incubate for 24 hours. The final test compound dose range was 30 µM to 14 nM in 200 µL of F12K medium containing 5% FBS, 0.3% DMSO.
[000406] On day three, the cell medium was removed and the cells were washed with 200 µL of PBS (phosphate buffered saline). Each reservoir was trypsinized to dissociate cells, then neutralized by adding 200 µL of complete F12K medium. Cells were pelleted and stained with an APC-conjugated mouse anti-human ICAM-1 antibody (CD54) (BD Pharmingen, San Diego, CA, Catalog #559771) for 20 minutes at 4°C. Cells were washed with buffer Ice-cold FACS (PBS + 2% FBS) and surface ICAM-1 expression was analyzed by flow cytometry. Detection was performed using a BD LSR I System Flow Cytometer purchased from BD Biosciences of San Jose, California. Events were enabled for live scattering and geometric mean was calculated (Becton-Dickinson CellQuest software version 3.3, Franklin Lakes, NJ). Geometric means were plotted against compound concentration to generate a dose-response curve. Example 10: Assay for ICAM1 Expression in IFNY Stimulated U937 Myeloid Cells
[000407] Human U937 monocytic cells up-regulate the surface expression of ICAM-1 (CD54) in response to a variety of different stimuli. Therefore, using ICAM-1 expression as readout, the compound's effects on different signaling pathways can be evaluated in the same cell type. IFNy upregulates ICAM-1 through activation of the JAK/Stat pathway. In this example, upregulation of ICAM-1 by IFNN was evaluated.
[000408] The human monocytic cell line U937 was obtained from ATCC Rockville Mariland, catalog number CRL-1593.2, and grown in RPM1-1640 medium containing 10% (v/v) FCS. U937 cells were grown in 10% RPMI. Cells were then plated at a concentration of 100,000 cells per 160 µL in 96-well flat-bottom plates. Test compounds were then diluted as follows: 10 mM test compound was diluted 1:5 in DMSO (3 µL test compound to 10 mM in 12 µL DMSO), followed by a 1:3 serial dilution of test compound in DMSO (6 µL of test compound serially diluted in 12 µL of DMSO to give 3-fold dilutions). Then 4 µL of test compound was transferred to 76 µL of 10% RPMI resulting in a 10X solution (100 µM of test compound, 5% DMSO). For the control reservoirs, 4 μL of DMSO was diluted in 76 μL of 10% RPMI. The assay was performed in duplicate with 8 points (8 concentrations at a 3-fold dilution starting at 10 µM) and with 4 wells with only DMSO (control reservoirs) under stimulated conditions and 4 wells with only DMSO under unstimulated conditions.
[000409] The diluted compound plate was mixed 2X using a multimek (Beckman Coulter of Brea, California) and then 20 µL of the diluted compounds were transferred to the 96 well plate containing 160 µL of cells, which were then remixed twice at low speeds. Cells and compounds were then pre-incubated for 30 minutes at 37°C with 5% CO 2 .
[000410] The 10X stimulation mix was made by preparing a 100 ng/ml solution of human IFNy in 10% RPMI. Cells and compound were then stimulated with 20 µl of IFNY stimulation mix to give a final concentration of 10 ng/ml IFNY, 10 µM test compound, and 0.5% DMSO. Cells were kept under conditions for stimulation for 18 to 24 hours at 37°C with 5% CO2.
[000411] Cells were transferred to a 96-well round bottom plate for staining and then kept on ice for the duration of the staining procedure. Cells were spun at 1000 rpm for 5 minutes at 4°C, after which the supernatant was removed. Following removal of the supernatant, 1 µL of APC-conjugated mouse anti-human ICAM-1 antibody was added per 100 µL of FACS buffer. Cells were then incubated on ice in the dark for 30 minutes. Following incubation, 150 µL of FACS buffer was added and cells were centrifuged at 1000 rpm for 5 minutes at 4°C, after which the supernatant was removed. After removing the supernatant, 200 µl of FACS buffer was added and the cells resuspended. After suspension, the cells were centrifuged at 1000 rpm for 5 min at 4°C. The supernatant was then removed before resuspending the cells in 150 µL of FACS buffer.
[000412] Detection was performed using a BD LSR I System Flow Cytometer purchased from BD Biosciences of San Jose, California. Live cells were activated to live scatter and the geometric mean of ICAM-APC was measured (Becton-Dickinson CellQuest software version 3.3, Franklin Lakes, NJ). Both % of live cells and ICAM-1 expression were analyzed. Assays for test compounds were performed in parallel with a control compound of known activity. The EC50 for the control compound is typically 40 to 100 nM. Example 11: Kinase Assays by Fluorescence Polarization of JAK1, JAK2 and JAK3
[000413] This assay can be used to determine the potency of a compound described herein against certain JAK kinases and the selectivity of a compound described herein in inhibiting certain JAK kinase activities in vitro. Reagents and buffers
[000414] Green Tyrosine Kinase Kit (Invitrogen, Cat# P2837)
[000415] Gamma Acetylated Bovine Globulin (BGG) (Invitrogen, Cat# P2255)
[000416] JAK1 Active (Carna Biosciences)
[000417] Active JAK2 (Carna Biosciences) Active JAK3 (Carna Biosciences)
[000418] TK2 Peptide (Biotin-EGPWLEEEEEAYGWMDF-CONH2) (SynPep Custom Synthesis) Methods
[000419] Test compounds were serially diluted in DMSO starting at 500x the desired final concentration and then diluted to 1% DMSO in kinase buffer (20 mM HEPES, pH 7.4, 5 mM MgCl2, 2 mM MnCl 2 , 1 mM DTT, 0.1 mg/ml acetylated BGG). Test compound in 1% DMSO (0.2% final DMSO) was mixed 1:5 with ATP and substrate in kinase buffer at room temperature.
[000420] Kinase reactions were performed in a final volume of 20 μL containing peptide substrate and ATP and started by adding kinase in kinase buffer. Reactions were allowed to process to room temperature. The final substrate, ATP and enzyme concentrations, and reaction times for the different kinase assays are listed in Table III. Table III

[000421] Reactions were stopped by addition of 20 μL of PTK quenched mixture containing EDTA/anti-phosphotyrosine antibody (1X final)/fluorescent phosphopeptide tracer (0.5X final) diluted in FP Dilution Buffer according to manufacturer's instructions (Invitrogen). Plates were incubated for 30 minutes in the dark at room temperature and then read on a Polarion fluorescence polarization plate reader (Tecan).
[000422] Data were converted to the amount of phosphopeptide present using a calibration curve generated by competition with the phosphopeptide competitor provided in the Tyrosine Kinase Assay Kit, Green (Invitrogen). For IC50 determination, compounds were tested at eleven concentrations in duplicate and curve fitting was performed by nonlinear regression analysis using Matlab version 6.5 (MathWorks, Inc., Natick, MA, USA). Example 12: Constitutively Active JAK2 Dependent Cell Proliferation Assays
[000423] A mutation in the JH2 pseudokinase domain of JAK2 (JAK2 V617F) has been described in chronic myeloproliferative disorders as well as a subset of acute myeloid leukemia (AML) cell lines. Mutation of the negative regulatory domain JH2 deregulates the kinase allowing it to constitutively associate with the EPO receptor and become activated. UKE-1 cells, derived from an AML patient, express JAK2 V617F which directs their proliferation. The IL-3 dependent BaF3 myeloid cell line was engineered to express JAK2 V617F allowing it to proliferate in an IL-3 independent manner. The effect of JAK inhibitors on the proliferation of these cell lines can be used to assess the cellular activity of compounds against JAK2. Reagents and Buffers
[000424] Dimethyl Sulfoxide (DMSO) (Sigma-Aldrich, Cat No D2650) (Control)
[000425] Iscove DMEM, ATCC Catalog #30-2005
[000426] HEPES 1 M, Cellgro Catalog #25-060-CI (100 ml)
[000427] 100 mM Sodium Pyruvate, Cellgro Catalog #25-000-CI (100 ml)
[000428] Penicillin/Streptomycin, 10000 U/ml each, Cellgro Catalog #30-0 02-CI (100 ml)
[000429] RPMI 1640 (Cellgro, Cat No 10-0 40-CM)
[000430] Fetal Bovine Serum (JRH, Cat No 12106-500M)
[000431] Donor Equine Serum, Hyclone Catalog #SH30074.02 (100 ml)
[000432] 50 μM Hydrocortisone Solution, Sigma Catalog #H6909-10 ml (10 ml) Culture Conditions
[000433] BaF3 V617F cells were maintained and plated in RPMI with 10% FBS. The plating density for these cells was 1 X 105 cells/ml.
[000434] UKE-1 were maintained and plated in Iscove's DMEM containing 10% FBS, 10% equine serum, 1% penicillin/streptomycin and 1 µM hydrocortisone. The plating density for these cells was 0.4 X 106 cells/ml Methods
[000435] Cells were resuspended in a medium corresponding to a required cell density (see above). 100 μ of cell suspension was added to each well of a white flat-bottomed 96-well plate. Compound was serially diluted in DMSO from 5 mM in 3-fold dilutions, and then diluted 1:250 in RPMI 1640 medium containing 5% FBS and pen/strep. 100 µL of the resulting 2X compound solution was added per well in duplicate and cells were allowed to proliferate for 72 hours at 37°C.
[000436] Proliferation was measured using Cell-Titer-Glo. The substrate was thawed and allowed to come to room temperature. After removing the top 100 µL of media from each well, 100 µL of the premixed Cell-Titer-Glo reagent was added to each well. Plates were mixed on an orbital shaker for three minutes to induce lysis and incubated at room temperature for an additional five minutes to allow the signal to equilibrate. Luminescence was read on the Wallac Plate Reader.
[000437] The results of the ability of the compounds described herein to inhibit JAK3 activity, when tested under conditions described in Example 3 above, are shown in Table IV below. The compound designations in Table IV are consistent with those in Tables I to II above. In Table IV the activity is indicated by the following ranges: “A” represents compounds having an IC50 < 0.5 μM; “B” represents compounds having an IC50 > 0.5 µM and < 5 µM; “C” represents compounds having an IC50 > 5 µM and < 10 µM; and “D” represents compounds having activity > 10 μM. A blank in Table IV indicates that the compound was not tested in the assay of Example 3. Table IV






[000438] Although the foregoing invention has been described in some details to facilitate understanding, the described embodiments are to be considered illustrative and not limiting. It will be evident to a person of skill in the art that certain changes and modifications may be practiced within the scope of the appended claims.

权利要求:
Claims (22)
[0001]
1. Compound, characterized by the fact that it is of the formula IA:
[0002]
2. Compound according to claim 1, characterized in that any of the conditions a) to j) are applied.
[0003]
3. Compound according to claim 1, characterized in that condition j) is applied.
[0004]
4. Compound according to claim 3, characterized in that R5 is H, halo or C1-6 alkyl.
[0005]
5. Compound according to claim 1, characterized in that condition h) is applied.
[0006]
6. Compound according to claim 1, condition b), characterized in that R2a is H; R2b is OC1-3 alkyl; R2c is alkyl-OC1-3; and R2d is F.
[0007]
7. Compound according to claim 1, characterized in that condition i) is applied.
[0008]
8. Compound according to claim 1, characterized in that condition k) is applied.
[0009]
9. Compound according to claim 1, characterized in that condition l) is applied.
[0010]
10. A compound according to claim 1, characterized in that it is: I-29: 5-[2-(2-fluoro-3-methoxy-4-methyl-phenylamino)-5-methyl-trifluoroacetate salt pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-50: 5-[2-(2-Fluoro-3,4-dimethoxy-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-51: 5-(5-chloro-2-(2-fluoro-3,4-dimethoxyphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-67: 5-(5-chloro-2-(2-fluoro-3-methoxy-4-methylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-68: 5-[2-(4-ethyl-2-fluoro-3-methoxy-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one trifluoroacetate salt; I-71: 5-(5-chloro-2-(4-ethyl-2-fluoro-3-methoxyphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-101: 5-(2-(3,4-diethoxy-2-fluorophenylamino)-5-methylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; II-8: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl) - N2-(1-oxo-1,3-dihydro-isobenzofuran -6-yl)-2,4-pyrimidinediamine; II-9: N2-(1,3-dihydro-isobenzofuran-5-yl)-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl ) -2,4-pyrimidinediamine; II-10: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(1-oxo-1 ,3-dihydro-isobenzofuran-5-yl)-2,4-pyrimidinediamine; II-11: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl) - N2-(1-oxo-1,3-dihydro-2H -isoindol-6-yl)-2,4-pyrimidinediamine; II-12: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl) - N2-(1-oxo-1,3-dihydro-7 -methylisobenzofuran-4-yl)-2,4-pyrimidinediamine; or II-13: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl) -N2-(1-oxo-1,3-dihydro- 4-methylisobenzofuran-6-yl)-2,4-pyrimidinediamine.
[0011]
11. Compound according to claim 1, characterized in that it is: I-129: alcohol trifluoroacetate salt 4-(4-(2,3-dihydro-2-oxobenzo[d]oxazol-5- ylamino)-5-methylpyrimidin-2-ylamino)-2-methoxy-6-methylbenzyl; I-142: N2-(3,5-dimethyl-4-hydroxymethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2 ,4-pyrimidinediamine; I-147: N2-(3,4-dimethyl-5-hydroxymethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2 ,4-pyrimidinediamine; I-154: N2-(4-fluoro-3-hydroxymethyl-5-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl) -2,4-pyrimidinediamine; I-225: 5-(2-(4-(hydroxymethyl)-3-methoxy-5-methylphenylamino)-6-D-5-trideuteromethylpyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; or I-230: 4-(4-(2,3-dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-methylpyrimidin-2-ylamino)-2-methoxy-6-methylbenzyl alcohol.
[0012]
12. Compound according to claim 1, characterized in that it has the structure
[0013]
13. Compound according to claim 1, characterized in that it has the structure
[0014]
14. A compound according to claim 1, characterized in that it is I-142: N2-(3,5-dimethyl-4-hydroxymethyl)phenyl-5-methyl-N4-(2-oxo-2,3- dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine.
[0015]
15. A compound according to claim 1, characterized in that it is I-147: N2-(3,4-dimethyl-5-hydroxymethyl)phenyl-5-methyl-N4-(2-oxo-2,3- dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine.
[0016]
16. A compound according to claim 11, characterized in that it is I-154: N2-(4-fluoro-3-hydroxymethyl-5-methyl)phenyl-5-methyl- N4-(2-oxo-2, 3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine.
[0017]
17. A compound, characterized in that it is: I-1: 5-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2 formate salt -ylamino)-2-methylbenzonitrile; I-2: 4-(4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide trifluoroacetate salt; I-3: 3-(4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide trifluoroacetate salt; I-4: 5-(5-chloro-2-(phenylamino)pyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-5: 5-(5-chloro-2-(3,4-dimethylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-6: 5-(5-chloro-2-(3,4,5-trimethylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-7: 5-(5-chloro-2-(2,4-difluoro-3-methoxyphenylamino)pyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-8: 5-(5-chloro-2-(3-chloro-5-fluorophenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt; I-9: 5-(5-chloro-2-(4-methyl-3-(trifluoromethyl)phenylamino)pyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one; I-10: 5-(5-chloro-2-(3,5-dimethylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-11: 4-(5-chloro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide trifluoroacetate salt; I-12: 5-(5-chloro-2-(3-methoxy-5-(trifluoromethyl)phenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-13: 5-(2-(3,5-difluorophenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-14: 5-(5-chloro-2-(3,5-difluorophenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-15: 5-(5-chloro-2-(4-(trifluoromethyl)phenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-16: 5-(5-bromo-2-(3,4,5-trimethylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt; I-17: 5-[2-(3-Dimethylamino-4-methyl-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-18: 5-[5-Ethynyl-2-(3,4,5-trimethyl-phenylamino)-pyrimidin-4-ylamino]-3H-benzooxazol-2-one trifluoroacetate salt; I-19: 5-[2-(2-Fluoro-3-methoxy-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-20: 5-(5-fluoro-2-(2-fluoro-3-methoxyphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-21: 5-(5-chloro-2-(2-fluoro-3-methoxyphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-22: 5-(5-bromo-2-(2-fluoro-3-methoxyphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-23: 5-(2-(2-fluoro-3-methoxyphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-24: 5-[2-(3-Dimethylamino-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one trifluoroacetate salt; I-25: 5-[2-(4-Dimethylamino-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-26: 5-[5-Methyl-2-(methyl-phenyl-amino)-pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-27: 5-[2-(4-Bromo-2-fluoro-3-methoxy-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-28: 5-[2-(4-Bromo-2-fluoro-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-30: {4-[5-Methyl-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidin-2-ylamino]-phenyl}-acetaldehyde; I-31: 5-[2-(3-Etynyl-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-32: 5-(5-chloro-2-(3-methoxy-4-methylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-33: 5-(5-chloro-2-(3-(dimethylamino)-4-methylphenylamino)pyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one; I-34: 5-[2-(3-Amino-4-methoxy-phenylamino)-5-methyl-pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-35: 5-[5-Methyl-2-(3,5-dimethyl-4-d3-methyl-phenylamino)-pyrimidin-4-ylamino]-3H-benzooxazol-2-one trifluoroacetate salt; I-36: 5-(5-chloro-2-(3,5-dimethyl-4-d3-methyl)phenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-37: 5-(5-fluoro-2-(3,5-dimethyl-4-d3-methyl)phenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-38: 5-(2-(3,5-dimethyl-4-d3-methyl)phenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-39: 4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-2-phenylamino-pyrimidine-5-carboxylic acid methyl ester trifluoroacetate salt; I-40: 4-(2-Oxo-2,3-dihydro-benzooxazol-5-ylamino)-2-(3,4,5-trimethyl-phenylamino)-pyrimidine-5-acid methyl ester trifluoroacetate salt carboxylic; I-41: 5-(5-nitro-2-(phenylamino)pyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-42: 5-(5-nitro-2-(3,4,5-trimethylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-43: 5-(2-(2,4-difluoro-3-methoxyphenylamino)-5-nitropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-44: 5-(2-(3-Methoxy-4-methylphenylamino)-5-nitropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-45: 5-(2-(3-(dimethylamino)-4-methylphenylamino)-5-nitropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-46: 5-(5-methyl-2-(3,5-dimethyl-4-d3-methyl)phenylamino)pyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one; I-47: 5-(2-(3-Methoxy-5-(trifluoromethyl)phenylamino)-5-nitropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-48: 2-(2,4-Difluoro-3-methoxy-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine-acid methyl ester trifluoroacetate salt 5-carboxylic acid; I-49: 4-(2-Oxo-2,3-dihydro-benzooxazol-5-ylamino)-2-(3,4,5-trimethyl-phenylamino)-pyrimidine-5-carboxylic acid trifluoroacetate salt; I-52: 5-(5-chloro-2-(3-methoxy-4,5-dimethylphenylamino)pyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one; I-53: 5-{2-[2-Fluoro-3-(2-methoxy-ethoxy)-4-methyl-phenylamino]-5-methyl-pyrimidin-4-ylamino}-3H-benzooxazol-2-one; I-54: 2-(4-Carbamoyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine-5-carboxylic acid methyl ester trifluoroacetate salt; I-55: 2-(3-Methoxy-5-trifluoromethyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine-5-acid methyl ester trifluoroacetate salt carboxylic; I-56: 2-(3-Methoxy-4-methyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine-5-acid methyl ester trifluoroacetate salt carboxylic; I-57: 2-(3-Dimethylamino-4-methyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine-5-acid methyl ester trifluoroacetate salt carboxylic; I-58: 2-(4-Fluoro-3-methoxy-5-methyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-acid methyl ester trifluoroacetate salt pyrimidine-5-carboxylic acid; I-59: 2-(3-Methoxy-4,5-dimethyl-phenylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine acid methyl ester trifluoroacetate salt 5-carboxylic acid; I-60: 4-(5-nitro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide trifluoroacetate salt; I-61: 5-(5-fluoro-2-(2-fluoro-3-(2-methoxyethoxy)-4-methylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) trifluoroacetate salt - one; I-62: 5-(2-(2-fluoro-3-(2-methoxyethoxy)-4-methylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-63: 5-[5-Hydroxymethyl-2-(3,4,5-trimethyl-phenylamino)-pyrimidin-4-ylamino]-3H-benzooxazol-2-one; I-64: 2,6-Dimethyl-4-[5-methyl-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidin-2-ylamino]-benzonitrile trifluoroacetate salt; I-65: 5-[5-Methyl-2-(3-vinyl-phenylamino)-pyrimidin-4-ylamino]-3H-benzooxazol-2-one trifluoroacetate salt; I-66: 5-(5-Chloro-2-(2-fluoro-3-(2-methoxyethoxy)-4-methylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) trifluoroacetate salt - one; I-69: 4-(5-chloro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-2,6-dimethylbenzonitrile] trifluoroacetate salt; I-70: 4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-2,6-dimethylbenzonitrile; I-72: 2-fluoro-3-(5-methyl-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzonitrile trifluoroacetate salt; I-73: 3-(5-chloro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-2-fluorobenzonitrile trifluoroacetate salt; I-74: methyl 2-(4-fluoro-3,5-dimethylphenylamino)-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidine-5-carboxylate trifluoroacetate salt ; I-75: 5-(2-(2-fluoro-4-methoxyphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-76: 5-(5-chloro-2-(2-fluoro-4-methoxyphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-77: 5-(2-(2-fluoro-3,4-bis(2-methoxyethoxy)-phenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(trifluoroacetate salt) 3H)-one; I-78: 5-(5-Chloro-2-(2-fluoro-3,4-bis(2-methoxyethoxy)-phenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) trifluoroacetate salt )-one; I-79: 5-(2-(4-fluoro-3,5-dimethylphenylamino)-5-(hydroxymethyl)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-80: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-(hydroxymethyl)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-81: 5-(5-(hydroxymethyl)-2-(3-methoxy-4,5-dimethyl-phenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-82: 5-(2-(3-(dimethylamino)-4,5-dimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt of; I-83: 5-(5-chloro-2-(3-(dimethylamino)-4,5-dimethyl-phenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-84: 5-(2-(3-(diethylamino)-4,5-dimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt; I-85: 5-(2-(3-(ethylamino)-4,5-dimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt; I-86: 5-(5-chloro-2-(3-(diethylamino)-4,5-dimethylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt; I-87: 5-(5-chloro-2-(3-(ethylamino)-4,5-dimethylphenylamino)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt; I-88: 5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-(benzo[d][1,3]dioxol-6-yl)benzo[ d]oxazol-2(3H)-one; I-89: 5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-((dimethylamino)methyl)benzo[d]oxazol-2(ditrifluoroacetate salt) 3H)-one; I-90: 7-((diethylamino)methyl)-5-(2-(3,4,5-trimethyl-phenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazole ditrifluoroacetate salt 2(3H)-one; I-91: 5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-((pyrrolidin-1-yl)methyl)benzo[d]oxazol-2( 3H)-one; I-92: 5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-((piperidin-1-yl)methyl)benzo[d]oxazol-2( 3H)-one; I-93: 5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-((4-methylpiperazin-1-yl)methyl)benzo[d]oxazol- 2(3H)-one; I-94: 4-((5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-2,3-dihydro-2-oxobenzo[d]oxazol-7- TERC-butyl yl)methyl)piperazine-1-carboxylate; I-95: 5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-((piperazin-1-yl)methyl)benzo[d] ditrifluoroacetate salt oxazol-2(3H)-one; I-96: 5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-((E)-3-chloroprop-1-enyl)benzo[d]oxazole -2(3H)-one; I-97: 5-(5-((diethylamino)methyl)-2-(3,4,5-trimethyl-phenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-ditrifluoroacetate salt one; I-98: 5-(2-(3,4,5-Trimethylphenylamino)-5-((pyrrolidin-1-yl)methyl)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-99: 5-(2-(3,4,5-Trimethylphenylamino)-5-((piperidin-1-yl)methyl)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-100: 5-(2-(3,4,5-Trimethylphenylamino)-5-((4-methylpiperazin-1-yl)methyl)-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) -one; I-102: 5-(2-(2-fluoro-3,4-d6-dimethoxyphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-103: 5-(2-(3-Ethoxy-4,5-dimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one ditrifluoroacetate salt; I-104: 5-(5-chloro-2-(3-ethoxy-4,5-dimethylphenyl-amino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one ditrifluoroacetate salt; I-105: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-106: 5-(2-(4-fluoro-3-methoxy-5-methyl-phenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazole-2(3H) methanesulfonic acid salt )-one; I-107: benzene sulfonic acid salt of 5-(2-(4-fluoro-3-methoxy-5-methyl-phenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) )-one; I-108: benzene sulfonic acid salt of 5-(2-(3-methoxy-4,5-dimethyl-phenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)- one; I-109: 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-110: 5-(5-fluoro-2-(3-methoxy-4,5-dimethylphenyl-amino)pyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-trifluoroacetate salt one; I-111: 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-112: 7-fluoro-5-(5-fluoro-2-(3-methoxy-4,5-dimethylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-113: 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one; I-114: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one sulfuric acid salt ; I-115: (5-(2-(3-methoxy-4,5-dimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-2-oxobenzo[d]oxazol-3(2H)-yl)methyl phosphate of disodium; I-116: 5-(2-(4-fluoro-3-hydroxy-5-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-117: 5-(2-(3-hydroxy-4,5-dimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-118: calcium phosphate dihydrogen salt of (5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)-2-oxobenzo[d]oxazol- 3(2H)-yl)methyl; I-119: 5-(5-fluoro-2-(2-fluoro-5-methoxyphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-120: 5-(2-(3-fluoro-5-methoxy-4-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-121: 5-(5-fluoro-2-(3-fluoro-5-methoxy-4-methylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-122: 5-(2-(2-fluoro-5-methoxy-4-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-123: 5-(5-fluoro-2-(2-fluoro-5-methoxy-4-methylphenylamino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-124: 5-(2-(4-chloro-3-methoxy-5-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-125: 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-3-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-126: 5-(2-(3,4,5-Trimethylphenylamino)-5-methyl-pyrimidin-4-ylamino)-3-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-127: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-methyl-pyrimidin-4-ylamino)-3-methylbenzo[d]oxazol-2(3H) trifluoroacetate salt - one; I-128: ethyl 4-(4-(2,3-dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-methyl-pyrimidin-2-ylamino)-2-methoxy-6-methylbenzoate; I-130: arginine salt of 5-methyl-N4-[3-(phosphonooxy)methyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl]-N2-(3,4, 5-trimethyl)phenyl-2,4-pyrimidinediamine; I-131: 5-Methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(3,4,5-trimethyl)phenyl benzene sulfonic acid salt -2,4-pyrimidinediamine; I-132: Tris salt 5-methyl-N4-[3-(phosphonooxy)methyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl]-N2-(3,4,5- -trimethyl)phenyl-2,4-pyrimidinediamine; I-133: N4-{3-[2-(N,N-dimethylamino)ethoxy]phosphinyloxymethyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl}-5-methyl-N2- (3,4,5-trimethyl)phenyl-2,4-pyrimidinediamine; I-134: N4-{3-bis[2-(N,N-dimethylamino)ethoxy]phosphinyloxymethyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl}-5-methyl-N2 -(3,4,5-trimethyl)phenyl-2,4-pyrimidine-diamine; I-135: 5-Methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(3,4,5-trimethyl)-phenyl pamoic acid salt -2,4-pyrimidinediamine; I-136: dipotassium salt of 5-methyl-N4-[3-(phosphonooxy)methyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl]-N2-(3,4,5- -trimethyl)phenyl-2,4-pyrimidinediamine; I-137: 5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(3,4,5-trimethyl)phenyl-methane sulfonic acid salt 2,4-pyrimidinediamine; I-138: N2-(3,5-dimethyl-4-methoxycarbonyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine; I-139: N2-ethyl-N2-(3,4,5-trimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2 ,4-pyrimidinediamine; I-140: N2-(4-carboxy-3,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine; I-141: N2-(4-benzyloxycarbonyl-3,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine; I-143: N2-(3,5-dimethyl-4-methoxymethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine; I-144: N2-(3,4-dimethyl-5-methoxycarbonyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine; I-145: N2-(3-carboxy-4,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine; I-146: benzene sulfonic acid salt N2-(3,5-dimethyl-4-fluoro)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl )-2,4-pyrimidinediamine; I-148: N2-(4-n-butyl-3-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine; I-149: N2-(4-bromo-3,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine; I-150: N2-(4-tert-butoxycarbonyl-3,5-dimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2 ,4-pyrimidinediamine; I-151: N2-(3,5-dimethyl-4-fluoro)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl) sulfuric acid salt )-2,4-pyrimidinediamine; I-152: N2-(3-carboxy-4-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine ; I-153: N2-(4-fluoro-3-methoxycarbonyl-5-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2 ,4-pyrimidinediamine; I-155: N2-(3-carboxy-4-fluoro-5-methyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2 ,4-pyrimidinediamine; I-156: 5-Methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-N2-(3,4,5-trimethyl)phenyl-hydrochloric acid salt 2,4-pyrimidinediamine; I-157: 5-(2-(4-acetylphenylamino)-5-fluoropyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one formate salt; I-158: 5-(2-(4-(1-(cyclopropylamino)ethyl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt; I-159: N-cyclobutyl-4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide formate salt; I-160: 4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-N-propylbenzamide formate salt; I-161: N-cyclopropyl-4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide formate salt; I-162: N-ethyl-4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzamide formate salt; I-163: 4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-N-isopropylbenzamide formate salt; I-164: N-cyclobutyl-4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-2-methoxybenzamide formate salt ; I-165: N-cyclopropyl-4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-2-( trifluoromethyl)benzamide; I-166: 4-(5-fluoro-4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)-N-phenyl-2-( trifluoromethyl)benzamide; I-167: 5-(2-(4-methyl-3-(methylsulfonyl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-168: 5-(2-(4-fluoro-3-(methylsulfonyl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-169: 5-(2-(3-fluoro-5-morpholinophenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one ditrifluoroacetate salt; I-170: 5-(2-(3-fluoro-5-(4-methylpiperazin-1-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H) ditrifluoroacetate salt -one; I-171: 3-(5-fluoro-4-(7-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzenesulfonamide trifluoroacetate salt; I-172: 5-(2-(3-(methylsulfonyl)phenylamino)-5-fluoro-pyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-173: 5-(2-(4-fluoro-3-(methylsulfonyl)phenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-174: 3-(5-fluoro-4-(7-fluoro-2-oxo-2,3-dihydro benzo[d]oxazol-5-ylamino)pyrimidin-2-ylamino)benzenesulfonamide trifluoroacetate salt; I-175: 5-(5-fluoro-2-(3-d3-methoxy-5-methylphenyl-amino)pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-176: 5-(2-(3-(trifluoromethyl)-5-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-177: 5-(2-(3-Methoxy-5-methylphenylamino)-5-fluoro-pyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-178: 5-(2-(4-methoxy-3-methylphenylamino)-5-fluoro-pyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-179: 5-(2-(3-Methoxy-5-methylphenylamino)-5-fluoro-pyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-180: 5-(2-(4-methoxy-3-methylphenylamino)-5-fluoro-pyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-181: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-182: 5-(2-(3,4-dimethoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-183: 5-(2-(3,4-dimethoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-184: 5-(2-(3,4-dimethoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-185: 5-(2-(3-chloro-4,5-dimethoxyphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-186: 5-(2-(4-(2-morpholinoethoxy)-3,5-dimethyl-phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-ditrifluoroacetate salt one; I-187: 5-(2-(3,5-dimethylphenylamino)-5-fluoro-pyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-188: 5-(2-(3,4,5-Trimethylphenylamino)-5-fluoro-pyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-189: 5-(2-(4-fluoro-3,5-dimethylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-190: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-191: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-5-fluoropyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt ; I-192: 5-(2-(2,4-Difluoro-3-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)-7-methylbenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-193: 5-(2-(2,4-difluoro-3-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)-7-fluorobenzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-194: 5-(2-(3,5-dimethoxy-4-methylphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-195: 5-(2-(3-Methoxy-4,5-dimethylphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-196: 5-(2-(4-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt; I-197: 5-(2-(3-(difluoromethyl)-4-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one formate salt; I-198: 5-(2-(3-(Difluoromethyl)-5-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-199: 5-(2-(3-(fluoromethyl)-5-methoxyphenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-200: 5-(2-(4-isopropylphenylamino)-5-fluoro-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-201: 5-(2-(4-tert-butylphenylamino)-5-fluoro-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-202: 5-(2-(p-tolylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; I-203: 5-(2-(4-methyl-3-(pyridin-4-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one; I-204: 5-(2-(4-methyl-3-(pyridin-3-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)-benzo[d]oxazol-2(3H)-one; I-205: 5-(2-(4-fluoro-3-(pyridin-4-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-206: 5-(2-(4-fluoro-3-(pyridin-3-yl)phenylamino)-5-fluoropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-207: 5-(2-(3-Methoxy-4-(pyridin-4-yl)phenylamino)-5-fluoro-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) formate salt -one; I-208: 5-(2-(4-methoxy-3-(pyridin-4-yl)phenylamino)-5-fluoro-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) formate salt -one; I-209: 5-(2-(4-methoxy-3-(pyridin-3-yl)phenylamino)-5-fluoro-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H) formate salt -one; I-210: 5-fluoro-N2-[3-(1-hydroxy-2,2,2-trifluoroethyl)]phenyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5- il)-2,4-pyrimidinediamine; I-211: 5-fluoro-N2-(3-methoxy-5-trifluoromethyl)phenyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine ; I-212: 5-fluoro-N2-(4-methoxy-3-trifluoromethyl)phenyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine ; I-213: N2-[3-(cyclopropylaminocarbonylmethoxy)-4-methoxy]phenyl-5-fluoro-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 - pyrimidinediamine; I-214: 5-fluoro-N2-(3,4,5-trimethyl)phenyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine ; I-215: N2-(3,5-dimethyl-4-fluoro)phenyl-5-fluoro-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4 -pyrimidinediamine; I-216: lysine salt of 5-methyl-N4-[3-(phosphonooxy)methyl-2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl]-N2-(3,4, 5-trimethyl)phenyl-2,4-pyrimidinediamine; I-217: N2-(3,4,5-Trimethyl)phenyl-5-methyl-N4-(2-oxo-2,3-dihydro-1,3-benzoxazol-5-yl)-2,4-pyrimidinediamine ; I-218: 5-(2-(2,4-difluoro-3-ethyleneoxyphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-219: 5-(2-(2,4-difluoro-3-ispropoxyphenylamino)-5-methyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-220: 5-(2-(4-(aminomethyl)phenylamino)-5-methyl-pyrimidin-4-ylamino)benzo-[d]oxazol-2(3H)-one; I-221: 5-(2-(4-fluoro-3-methoxy-5-methylphenylamino)-6-D-5-trideuteromethyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-222: 5-(2-(4-fluoro-3-trideuteromethoxy-5-methylphenylamino)-6-D-5-trideuteromethyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-223: 5-(2-(3-methoxy-4,5-dimethylphenylamino)-6-D-5-trideuteromethyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-224: 5-(2-(3-trideuteromethoxy-4,5-dimethylphenylamino)-6-D-5-trideuteromethyl-pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; I-226: 5-(2-(4-fluoro-3-trideuteromethoxy-5-methylphenyl-amino)-6-D-5-trideuteromethyl-pyrimidin-4-ylamino)-2-oxobenzo di-tert-butyl phosphate [d]oxazol-3(2H)-yl)methyl; I-227: 5-(2-(4-fluoro-3-trideuteromethoxy-5-methylphenylamino)-6-D-5-trideuteromethyl-pyrimidin-4-ylamino)-2-oxobenzo[d] tert-butyl hydrogen phosphate -oxazol-3(2H)-yl)methyl; I-228: 5-(2-(4-fluoro-3-trideuteromethoxy-5-methyl-phenylamino)-6-D-5-trideuteromethyl-pyrimidin-4-ylamino)-2-oxobenzo[d]-dihydrogen phosphate oxazol-3(2H)-yl)methyl; I-229: (5-(2-(4-fluoro-3-trideuteromethoxy-5-methylphenylamino)-6-D-5-trideuteromethyl-pyrimidin-4-ylamino)-2-oxobenzo[d]oxazol-3(2H) disodium )-yl)methyl phosphate; I-231: 4-(4-(2,3-dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-methyl-pyrimidin-2-ylamino)-2-methoxy-6-methylbenzoic acid.
[0018]
18. A compound, characterized in that it is: II-1: 5-(5-fluoro-2-(1-oxo-1,2,3,4-tetrahydro-isoquinolin-6-ylamino)pyrimidine trifluoroacetate salt -4-ylamino)benzo[d]oxazol-2(3H)-one; II-2: 5-(5-fluoro-2-(7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulen-2-ylamino) diformate salt pyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one; II-3: 5-(5-fluoro-2-(7-oxo-6,7,8,9-tetrahydro-5H-benzo[7]anulen-2-ylamino)pyrimidin-4-ylamino) diformate salt benzo[d]oxazol-2(3H)-one; II-4: (Z)-5-(5-fluoro-2-(2-methyl-1-oxo-1,2,3,6-tetrahydro benzo[c]azocin-9-ylamino)pyrimidin formate salt -4-ylamino)benzo[d]oxazol-2(3H)-one; II-5: 5-(5-fluoro-2-(5-oxo-6,7,8,9-tetrahydro-5H-benzo[7]anulen-2-ylamino)pyrimidin-4-ylamino) formate salt benzo[d]oxazol-2(3H)-one; II-6: 5-(2-(naphthalen-2-ylamino)-5-nitropyrimidin-4-ylamino)benzo[d]oxazol-2(3H)-one trifluoroacetate salt; II-7: 2-(Naphthalen-2-ylamino)-4-(2-oxo-2,3-dihydro-benzooxazol-5-ylamino)-pyrimidine-5-carboxylic acid methyl ester trifluoroacetate salt; II-23: 5-(5-fluoro-2-(1-oxo-2,3-dihydro-1H-benzo[c]azepin-7-ylamino)pyrimidin-4-ylamino)benzo[d] formate salt oxazol-2(3H)-one; II-24: 5-(5-fluoro-2-(2-methyl-1-oxo-2,3-dihydro-1H-benzo[c]azepin-7-ylamino)pyrimidin-4-ylamino) formate salt benzo[d]oxazol-2(3H)-one; II-25: 7-(4-(2,3-dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-fluoropyrimidin-2-ylamino)-2H-benzo[b][1]formate salt ,4]oxazin-3(4H)-one; II-26: 6-(4-(2,3-dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-fluoropyrimidin-2-ylamino)-3,4-dihydroquinolin-2(trifluoroacetate salt) 1H)-one; II-27: 7-(4-(2,3-dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-fluoropyrimidin-2-ylamino)-4,5-dihydro-1H- trifluoroacetate salt benzo[b]azepin-2(3H)-one; or II-28: 7-(4-(2,3-dihydro-2-oxobenzo[d]oxazol-5-ylamino)-5-fluoropyrimidin-2-ylamino)-4,5-dihydro-1 trifluoroacetate salt - methyl-1H-benzo[b]azepin-2(3H)-one.
[0019]
19. Pharmaceutical composition, characterized in that it comprises a compound as defined in any one of claims 1 to 18.
[0020]
20. A method of making a compound of the formula IA as defined in claim 1, characterized in that it comprises: (i) reacting a guanidine of the formula A-11 with a 1,3-dielectrophil to make a 4-hydroxy-pyrimidin- 2-yl-amine of the formula A-13;
[0021]
21. Method according to claim 20, characterized in that the 1,3-dielectrophil is an e-aldehyde ester of formula A-12:
[0022]
22. A compound as defined in any one of claims 1 to 18, or a pharmaceutical composition as defined in claim 19, characterized in that it is for use in a method of treating a T-cell mediated autoimmune disease, an allograft transplant rejection in a transplant recipient, a type IV hypersensitivity reaction or an eye disease and/or disorder. I I R3 R4 I where: I

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法律状态:
2018-01-23| B07D| Technical examination (opinion) related to article 229 of industrial property law [chapter 7.4 patent gazette]|

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2018-04-10| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|

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2019-07-02| B07E| Notification of approval relating to section 229 industrial property law [chapter 7.5 patent gazette]|Free format text: NOTIFICACAO DE ANUENCIA RELACIONADA COM O ART 229 DA LPI |

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2019-08-06| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

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2020-08-18| B06A| Patent application procedure suspended [chapter 6.1 patent gazette]|

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2021-02-23| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2021-05-25| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 27/07/2011, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

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申请号 | 申请日 | 专利标题

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US36857010P| true| 2010-07-28|2010-07-28|

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US61/368570|2010-07-28|

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PCT/US2011/045609|WO2012015972A1|2010-07-28|2011-07-27|Compositions and methods for inhibition of the jak pathway|

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