Bis(fluoroalkyl)-1,4-benzodiazepinone compounds, combination and use of said compounds

专利摘要:
BIS(FLUOROALKYL)-1,4-BENZODIAZEPINONE COMPOUNDS, THEIR PHARMACEUTICAL COMPOSITION, THEIR COMBINATION AND THEIR USE. The invention relates to compounds of Formula (I) or prodrugs thereof; (I) wherein: R1 is -CH2CF3 or -CH2CH2CF3; R2 is -CH2CF3, -CH2CH2CF3, or -CH2CH2CH2CF3; R3 is H or -CH3; each Ra is independently F, Cl, -CN, -OCH3, and/or -NHCH2CH2OCH3; and z is zero, 1, or 2. It likewise refers to methods of using such a compound to inhibit the Notch receptor, and pharmaceutical compositions comprising such compounds. These compounds are useful in treating, preventing, or delaying the progress of diseases or disorders in a variety of therapeutic areas, such as cancer.
公开号:BR112013024059B1
申请号:R112013024059-8
申请日:2012-03-22
公开日:2022-01-25
发明作者:Claude Quesnelle；Soong-Hoon Kim；Francis Y. Lee；Ashvinikumar Gavai
申请人:Bristol-Myers Squibb Company；
IPC主号:

专利说明:

DESCRIPTION
[0001] The present invention relates generally to benzodiazepinone compounds useful as Notch inhibitors. The invention also relates to pharmaceutical compositions comprising at least one compound according to the invention which is useful for the treatment of conditions related to the Notch reaction series, such as cancer and other proliferative diseases.
[0002] Notch signaling has been implicated in a variety of cellular processes, such as cell fate specification, differentiation, proliferation, apoptosis, and angiogenesis. (Bray, Nature Reviews Molecular Cell Biology, 7:678-689 (2006 ); Fortini, Developmental Cell 16:633-647 (2009 )). Notch proteins are single-pass heterodimeric transmembrane molecules. The Notch family includes 4 receivers, NOTCH 1-4, which become activated by binding to the ligands of the DSL family (Delta-similar 1, 3, 4 and Jagged 1 and 2).
[0003] Activation and maturation of NOTCH requires a series of processing steps, including a proteolytic cleavage step mediated by gamma-secretase, a multiprotein complex containing Presenilin 1 or Presenilin 2, nicastrin, APH1, and PEN2. As soon as NOTCH is cleaved, the intracellular domain of NOTCH (NICD) is released from the membrane. The released NICD translocates to the nucleus where it functions as a transcriptional activator in concert with members of the CSL family (RBPSUH, "hairless suppressor", and LAG1). NOTCH target genes include members of the HES family, such as HES-1. HES-1 functions as transcriptional repressors for genes such as HERP1 (also known as HEY2), HERP2 (also known as HEY1), and HATH1 (also known as ATOH1).
[0004] Aberrant activation of the Notch reaction series contributes to tumorigenesis. Activation of Notch signaling has been implicated in the pathogenesis of several solid tumors that include ovarian, pancreatic, as well as breast cancer and hematologic tumors such as leukemias, lymphomas, and multiple myeloma. The role of Notch inhibition and its utility in the treatment of various solid and hematologic tumors are described in Miele, L. et al., Current Cancer Drug Targets, 6:313-323 (2006 ); Bolos, V. et al., Endocrine Reviews, 28:339363 (2007); Shih, I.-M. and another, Cancer Research, 67:1879-1882 (2007 ); Yamaguchi, N. et al., Cancer Research, 68:1881-1888 (2008 ); Miele, L., Expert Review Anti-cancer Therapy, 8:1197-1201 (2008); Purew, B., Current Pharmaceutical Biotechnology, 10:154-160 (2009 ); Nefedova, Y. et al., Drug Resistance Updates, 11:210-218 (2008 ); Dufraine, J. et al., Oncogene, 27:5132-5137 (2008 ); and Jun, H.T. and another, Drug Development Research, 69:319-328 (2008 ).
[0005] There remains a need for compounds which are useful as Notch inhibitors and which have sufficient metabolic stability to provide effective levels of drug exposure. Furthermore, there remains a need for compounds useful as Notch inhibitors that can be orally or intravenously administered to a patient.
[0006] U.S. Patent At the. 7,053,084 B1 describes succinoylamino benzodiazepine compounds useful for treating neurological disorders such as Alzheimer's Disease. The reference describes that these sucinoylamino benzodiazepine compounds inhibit gamma secretase activity and the amyloid precursor protein process linked to the formation of neurological amyloid protein deposits. The reference does not describe the use of these compounds in the treatment of proliferative diseases such as cancer.
[0007] Applicants have found potent compounds that have activity as Notch inhibitors and have sufficient metabolic stability to provide effective drug exposure levels upon intravenous or oral administration. These compounds are provided to be useful as pharmaceuticals with desirable stability, bioavailability, therapeutic index, and toxicity values that are important to their pharmacology. SUMMARY OF THE INVENTION
[0008] The present invention satisfies the foregoing need by providing bis(fluoroalkyl)-1,4-benzodiazepinone compounds which are useful as selective inhibitors of the Notch signaling reaction series, including prodrugs thereof.
[0009] The present invention also provides pharmaceutical compositions comprising a pharmaceutically acceptable carrier; and at least one compound of Formula (I) or prodrugs thereof.
[00010] The present invention also provides a method of treating a disease or disorder associated with Notch receptor activity, the method comprising administering to a mammalian patient a compound of Formula (I) or pharmaceutically acceptable prodrugs thereof.
[00011] The present invention also provides processes and intermediates for preparing the compounds of Formula (I) or prodrugs thereof.
[00012] The present invention also provides the compounds of Formula (I), or prodrugs thereof, for use in therapy.
[00013] The present invention also provides the use of compounds of Formula (I), or prodrugs thereof, for the manufacture of a medicament for the treatment of cancer.
[00014] The compounds of Formula (I) and compositions comprising the compounds which are Notch inhibitors can be used in the treatment, prevention or cure of various conditions related to the Notch receptor. Pharmaceutical compositions comprising these compounds are useful in treating, preventing, or slowing the progress of diseases or disorders in a variety of therapeutic areas, such as cancer.
[00015] These and other aspects of the invention will be mentioned in expanded form as the description continues. BRIEF DESCRIPTION OF THE DRAWINGS
[00016] The invention is illustrated by reference to the accompanying drawings described below.
[00017] Figure 1 shows the experimental (at approximately 25 °C) and simulated (at approximately 25 °C) (CuKα λ=1.5418Â) PXRD patterns of Form N-1 of the compound of Example 1.
[00018] Figure 2 shows the experimental (at approximately 25 °C) and simulated (at approximately 25 °C) (CuKα λ =1.5418Â) PXRD patterns of Form A-2 of the compound of Example 1.
[00019] Figure 3 shows the experimental (at approximately 25 °C) and simulated (at approximately 25 °C) (CuKα λ =1.5418Â) PXRD patterns of Form EA-3 of the compound of Example 1.
[00020] Figure 4 shows the experimental (at approximately 25 °C) and simulated (at approximately -50 °C) (CuKα λ =1.5418Â) PXRD patterns of Form THF-2 of the compound of Example 1.
[00021] Figure 5 shows the experimental (at approximately 25 °C) and simulated (at approximately 25 °C) (CuKα λ =1.5418Â) PXRD patterns of Form M2-1 of the compound of Example 2.
[00022] Figure 6 shows the antitumor efficacy of Example 1 against TALL1 T-cell acute lymphoblastic leukemia. Each symbol represents the median tumor burden of a group of 8 mice. (•) Control; (■) Example 1, 5 mg/kg/adm, QD x 3, IV.
[00023] Figure 7 shows the in vivo antitumor activity of Example 2 on the TALL1 T-cell acute lymphoblastic leukemia cell line. Each symbol represents the median tumor burden of a group of 8 mice. (•) control; (Δ) Example 2, 12 mg/kg, QDx15; (■) Example 2, 6 mg/kg, QDx15; ( ) Example 2, 3 mg/kg, QDx15; (▲) Example 2, 1.5 mg/kg, QDx15; (o) Example 2, 0.75 mg/kg, QDx15.
[00024] Figure 8 shows the in vivo antitumor activity of Example 2 on the human breast carcinoma cell line MDA-MB-157. Each symbol represents the median tumor burden of a group of 8 mice. (the control; (▲) Example 2, 24 mg/kg; (■) Example 2, 18 mg/kg; (•) Example 2, 12 mg/kg.
[00025] Figure 9 shows the synergistic antitumor efficacy by chemotherapy combined with Example 1 and dasatinib in ALL-SIL T-cell lymphoblastic leukemia. Each symbol represents the median tumor burden of a group of 8 mice. (•) control; (♦) Example 1, 3.75 mg/kg/adm, QD x 3, weekly for 7 weeks, PO; (■) dasatinib, 10 mg/kg/adm, QD x 49, PO; (□) Example 1, 3.75 mg/kg/adm, QD x 3, weekly for 7 weeks, PO + dasatinib 10 mg/kg/adm, QD x 49, PO. When given on the same day, the two agents were given more or less simultaneously (Example 1 dasatinib preceded by less than 1 hr).
[00026] Figure 10 shows the synergistic antitumor efficacy by chemotherapy combined with Example 1 and dasatinib in ALL-SIL T-cell lymphoblastic leukemia. Each symbol represents the median tumor burden of a group of 8 mice. (•) control; (♦) Example 1, 7.5 mg/kg/adm QD x 3, weekly for 7 weeks, PO; (■) dasatinib, 10 mg/kg/adm, QD x 49, PO; (□) Example 1, 7.5 mg/kg/adm, QD x 3, weekly for 7 weeks, PO + dasatinib 10 mg/kg/adm, QD x 49, PO. When given on the same day, the two agents were given more or less simultaneously (Example 1 dasatinib preceded by less than 1 hr).
[00027] Figure 11 shows the synergistic antitumor efficacy by chemotherapy combined with Example 1 and Paclitaxel in MDA-MB-468 Human Breast Carcinoma. Each symbol represents the median tumor burden of a group of 8 mice. (•) control; (♦) Paclitaxel, 12 mg/kg/adm, Q7D x 6, IV; (▲) Example 1, 3.75 mg/kg/adm, QD x 3, weekly for 7 weeks, PO; (□) Combination of Example 1 and Paclitaxel.
[00028] Figure 12 shows the synergistic antitumor efficacy by chemotherapy combined with Example 1 and Paclitaxel in MDA-MB-468 Human Breast Carcinoma. Each symbol represents the median tumor burden of a group of 8 mice. (•) control; (♦) Paclitaxel, 12 mg/kg/adm, Q7D x 6, IV; (▲) Example 1, 7.5 mg/kg/adm, QD x 3, weekly for 7 weeks, PO; (□) Combination of Example 1 and Paclitaxel.
[00029] Figure 13 shows the synergistic antitumor efficacy by chemotherapy combined with Example 1 and Tamoxifen in MCF7 Human Breast Carcinoma. Each symbol represents the median tumor burden of a group of 8 mice. (•) control; (■) Tamoxifen, 20 mg/kg/adm, Q2D x 12, IP; (♦) Example 1, 3.75 mg/kg/adm, QD x 3, weekly for 3 weeks, PO; (□) Combination of Example 1 and Tamoxifen.
[00030] Figure 14 shows the synergistic antitumor efficacy by chemotherapy combined with Example 1 and Tamoxifen in MCF7 Human Breast Carcinoma. Each symbol represents the median tumor burden of a group of 8 mice. (•) control; (■) Tamoxifen, 20 mg/kg/adm, Q2D x 12, IP; (♦) Example 1, 7.5 mg/kg/adm, QD x 3, weekly for 3 weeks, PO; (□) Combination of Example 1 and Tamoxifen.
[00031] Figure 15 shows synergistic antitumor efficacy by chemotherapy combined with Example 1 and dexamethasone (Dexa) in HPB-ALL human T-ALL leukemia xenografts. Each symbol represents the median tumor burden of a group of 6-8 mice. (•) control; (♦) dexamethasone, 7.5 mg/kg/adm, QD x 14, IP; (Δ) Example 1, 3.75 mg/kg/adm, QD x 3, weekly for 3 weeks, PO; (□) Combination of Example 1 and dexamethasone.
[00032] Figure 16 shows the synergistic antitumor efficacy by chemotherapy combined with Example 1 and carboplatin in PA-1 terato-human ovarian carcinoma. Each symbol represents the median tumor burden of a group of 8 mice. (•) control; (□) carboplatin, 90 mg/kg/adm, Q7D x 3, IV; (Δ) Example 1, 1 mg/kg/adm, QD x 21, PO; (♦) Combination of Example 1 and carboplatin. DETAILED DESCRIPTION
[00033] The first aspect of the present invention provides the compounds of Formula (I):

[00034] or prodrugs thereof; in
[00035] R1 is -CH2CF3 or -CH2CH2CF3;
[00036] R2 is -CH2CF3, -CH2CH2CF3, or -CH2CH2CH2CF3;
[00037] R3 is H or -CH3;
[00038] each Ra is independently F, Cl, -CN, -OCH3, and/or -NHCH2CH2OCH3; and
[00039] z is zero, 1, or 2.
[00040] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CF3; and R2, R3, Ra, and z are defined in the first aspect. Included in this embodiment are compounds wherein R2 is -CH2CF3 or -CH2CH2CF3.
[00041] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CH2CF3; and R2 , R3 , Ra , and z are defined in the first aspect. Included in this embodiment are compounds wherein R2 is -CH2CF3 or -CH2CH2CF3.
[00042] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R2 is -CH2CF3; and R1, R3, Ra, and z are defined in the first aspect. Included in this embodiment are compounds wherein R1 is -CH2CH2CF3.
[00043] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R2 is -CH2CH2CF3; and R1, R3, Ra, and z are defined in the first aspect. Included in this embodiment are compounds wherein R1 is -CH2CH2CF3.
[00044] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R2 is -CH2CH2CH2CF3; and R1, R3, Ra, and z are defined in the first aspect. Included in this embodiment are compounds wherein R1 is -CH2CH2CF3.
[00045] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R3 is H; and R1, R2, Ra, and z are defined in the first aspect. Included in this embodiment are compounds wherein R1 is deuterium (D) or tritium (T). Also included in this embodiment are compounds wherein R1 is -CH2CH2CF3 and R2 is -CH2CH2CF3.
[00046] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R3 is -CH3; and R1, R2, Ra, and z are defined in the first aspect. R3 includes methyl groups in which one or more hydrogen atoms are isotopically substituted with deuterium (D) and/or tritium (T). In an example of this embodiment, R3 is -CD3. Also included in this embodiment are compounds wherein R1 is -CH2CH2CF3 and R2 is -CH2CH2CF3.
[00047] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein z is 2 and each Ra is independently F, Cl, -CN, -OCH3, and/or -NHCH2CH2OCH3; and R1, R2, and R3 are defined in the first aspect. Included in this embodiment are compounds wherein R1 is -CH2CH2CF3 and R2 is -CH2CH2CF3.
[00048] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein z is 1 and Ra is F, Cl, -CN, -OCH3, or -NHCH2CH2OCH3; and R1, R2, and R3 are defined in the first aspect. Included in this embodiment are compounds wherein R1 is -CH2CH2CF3 and R2 is -CH2CH2CF3.
[00049] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein z is zero; and R1, R2, and R3 are defined in the first aspect. Included in this embodiment are compounds wherein R1 is -CH2CH2CF3 and R2 is -CH2CF3 or -CH2CH2CF3.
[00050] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CF3; R2 is -CH2CF3; R3 is H or -CH3; and z is zero.
[00051] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CF3; R2 is -CH2CH2CF3; R3 is H or -CH3; and z is zero.
[00052] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CF3; R2 is -CH2CH2CH2CF3; R3 is H or -CH3; and z is zero.
[00053] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CH2CF3; R2 is -CH2CF3; R3 is H or -CH3; and z is zero.
[00054] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CH2CF3; R2 is -CH2CH2CF3; R3 is H or -CH3; and z is zero.
[00055] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CH2CF3; R2 is -CH2CH2CH2CF3; R3 is H or -CH3; and z is zero.
[00056] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CF3; R2 is -CH2CF3; R3 is H or -CH3; z is 1; and Ra is F, Cl, -CN, -OCH3, and/or -NHCH2CH2OCH3.
[00057] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CF3; R2 is -CH2CH2CF3; R3 is H or -CH3; z is 1; and Ra is F, Cl, -CN, -OCH3, and/or -NHCH2CH2OCH3.
[00058] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CF3; R2 is -CH2CH2CH2CF3; R3 is H or -CH3; z is 1; and Ra is F, Cl, -CN, -OCH3, and/or -NHCH2CH2OCH3.
[00059] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CH2CF3; R2 is -CH2CF3; R3 is H or -CH3; z is 1; and Ra is F, Cl, -CN, -OCH3, and/or -NHCH2CH2OCH3.
[00060] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CH2CF3; R2 is -CH2CH2CF3; R3 is H or -CH3; z is 1; and Ra is F, Cl, -CN, -OCH3, and/or -NHCH2CH2OCH3.
[00061] One embodiment provides a compound of Formula (I) or prodrugs thereof, wherein R1 is -CH2CH2CF3; R2 is -CH2CH2CH2CF3; R3 is H or -CH3; z is 1; and Ra is F, Cl, -CN, -OCH3, and/or -NHCH2CH2OCH3.
[00062] One embodiment provides a compound according to claim 1 or prodrugs thereof, selected from:


[00063] One embodiment provides a compound according to claim 1 or prodrugs thereof, selected from:

[00064] One embodiment provides a compound according to claim 1 or prodrugs thereof, selected from:

[00065] One embodiment provides claim 1 or prodrugs thereof, selected from:

[00066] One embodiment provides a compound of Formula (I) selected from: (2R,3S)-N-((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro -1H-1,4-benzodiazepin-3-yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide (2R,3S)-N-((3S)-2-oxo-5-phenyl- 2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide (2); (2R,3S)-N-((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2-(2 ,2,2-trifluoroethyl)-3-(3,3,3-trifluoropropyl)succinamide (3); (2R,3S)-N-((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-3-(2 ,2,2-trifluoroethyl)-2-(3,3,3-trifluoropropyl)succinamide (4); (2R,3S)-N-((3S)-1-(2H3)methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2 ,3-bis(3,3,3-trifluoropropyl)succinamide (5); (2R,3S)-N-((3S)-7-chloro-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (6); (2R,3S)-N-((3S)-8-methoxy-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (7); (2R,3S)-N-((3S)-8-fluoro-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (8); (2R,3S)-N-((3S)-7-methoxy-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (9); (2R,3S)-N-((3S)-7-fluoro-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (10); (2R,3S)-N-((3S)-8-chloro-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (11); (2R,3S)-N-((3S)-9-methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3- bis(3,3,3-trifluoropropyl)succinamide (12); (2R,3S)-N-((3S)-8-methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3- bis(3,3,3-trifluoropropyl)succinamide (13); (2R,3S)-N-((3S)-7-methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3- bis(3,3,3-trifluoropropyl)succinamide (14); (2R,3S)-N-((3S)-8-cyano-9-methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (15); (2R,3S)-N-((3S)-8,9-dichloro-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2, 3-bis(3,3,3-trifluoropropyl)succinamide (16); (2R,3S)-N-((3S)-9-fluoro-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3- bis(3,3,3-trifluoropropyl)succinamide (17); (2R,3S)-N-((3S)-9-chloro-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3- bis(3,3,3-trifluoropropyl)succinamide (18); (2R,3S)-N-((3S)-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-3-(4,4,4 -trifluorobutyl)-2-(3,3,3-trifluoropropyl)succinamide (19); (2R,3S)-N1-((3S)-8-methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-3-(4 ,4,4-trifluorobutyl)-2-(3,3,3-trifluoropropyl)succinamide (20); and (2R,3S)-N-((3S)-9-((2-methoxyethyl)amino)-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3 -yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide (21); and prodrugs of one or more of the above compounds.
[00067] The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. This invention encompasses all combinations of the aspects and/or embodiments of the invention noted herein. It is understood that any and all embodiments of the present invention may be considered in conjunction with any other embodiment or embodiments to describe the addition of further embodiments. It is likewise to be understood that each individual element of the modalities is meant to be combined with any and all other elements of any modality to describe an additional modality. DEFINITIONS
[00068] The aspects and advantages of the invention can be more easily understood by those of ordinary skill in the art by reading the following detailed description. It should be appreciated that certain aspects of the invention which are, for the sake of clarity, described above and below in the context of separate embodiments, may likewise be combined to form a single embodiment. Conversely, various aspects of the invention which are, for the sake of brevity, described in the context of a single embodiment, may be similarly combined to form subcombinations thereof. Embodiments identified herein as exemplary or preferred are intended to be illustrative and limiting.
[00069] Unless specifically stated otherwise herein, references made to the singular may likewise include the plural. For example, "a" and "an" can refer to either of the two, or one or more.
[00070] Unless otherwise indicated, any heteroatom with unsatisfied valences is assumed to have enough hydrogen atoms to satisfy the valences.
[00071] The definitions mentioned herein take precedence over the definitions mentioned in any patent, patent application and/or patent application publication incorporated herein by reference.
[00072] Listed below are definitions of various terms to describe the present invention. These definitions apply to terms as they are used throughout the descriptive report (unless they are otherwise limited in specific examples) individually or as part of a larger group.
[00073] Throughout the specification, groups and substituents thereof may be chosen by one skilled in the field to provide stable and compound moieties.
[00074] According to a convention used in the art,
is used in structural formulas herein to describe the bond that is the point of attachment of either the moiety or substituent to the backbone structure or core.
[00075] The terms "halo" and "halogen" when used herein refer to F, Cl, Br, or I.
[00076] The term "alkyl" when used herein also refers to saturated aliphatic straight and branched chain hydrocarbon groups containing, for example, from 1 to 12 carbon atoms, from 1 to 6 carbon atoms, and from 1 to 4 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g. n-propyl and i-propyl), butyl (e.g. n-butyl, i-butyl, sec -butyl, and t-butyl), and pentyl (e.g., n-pentyl, isopentyl, neopentyl), n-hexyl, 2-methylpentyl, 2-ethylbutyl, 3-methylpentyl, and 4-methylpentyl. When numbers appear in a subscription after the symbol "C", the subscription more specifically defines the number of carbon atoms that a particular group can contain. For example, "C1-6alkyl" denotes straight and branched chain alkyl groups of one to six carbon atoms.
[00077] The phrase "pharmaceutically acceptable" is employed to refer to those compounds, materials, compositions and/or dosage forms that are within the scope of safe medical judgment, suitable for use in contact with the tissues of humans and animals. without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
[00078] Compounds of Formula (I) may be supplied as amorphous solids or crystalline solids. Lyophilization may be employed to provide compounds of Formula (I) as a solid.
[00079] Any compound that can be converted in vivo to provide the bioactive agent (i.e., the compound of Formula (I)) is a prodrug within the scope and spirit of the invention.
[00080] Various forms of prodrugs are well known in the art and are described in: a) Wermuth, C.G. and another, The Practice of Medicinal Chemistry, Chapter 31, Academic Press (1996); b) Bundgaard, H. ed., Design of Prodrugs, Elsevier (1985); c) Bundgaard, H., Chapter 5, "Design and Application of Pro-drugs," Krosgaard-Larsen, P. et al., eds., A Textbook of Drug Design and Development, pp. 113-191, Harwood Academic Publishers (1991); and d) Testa, B. et al., Hydrolysis in Drug and Prodrug Metabolism, Wiley-VCH (2003).
[00081] Furthermore, the compound of Formula (I) is, subsequent to its preparation, preferably isolated and purified to obtain a composition which contains an amount by weight equal to or greater than 99% of a compound of Formula (I) ( "substantially pure") which is then used or formulated as described herein. Such "substantially pure" compounds of Formula (I) are likewise contemplated herein as part of the present invention.
[00082] "Stable compound" and "stable structure" are intended to indicate a compound that is strong enough to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an effective therapeutic agent. The present invention is intended to incorporate the stable compounds.
[00083] "Therapeutically effective amount" is intended to include an amount of a compound of the present invention alone or an amount of the combination of claimed compounds or an amount of a compound of the present invention in combination with other active ingredients effective to act as an inhibitor for a NOTCH receptor, or effective to treat or prevent proliferative diseases such as cancer.
[00084] When used herein, "treating" or "treatment" encompasses treating a disease state in a mammal, particularly a human, and includes: (a) preventing the disease state from occurring in a mammal, in particular, when such a mammal is predisposed to the disease state, however, has not yet been diagnosed as having it; (b) inhibit the disease state, ie, stop its development; and/or (c) alleviating the disease state, i.e., causing the disease state to regress.
[00085] The compounds of the present invention are intended to include all isotopes of atoms occurring in the present compounds. Isotopes include those atoms that have the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include deuterium (D) and tritium (T). Carbon isotopes include 13C and 14C. Isotopically labeled compounds of the invention generally can be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically labeled reagent in place of the otherwise unlabeled reagent employed. Crystal Forms of Compound of Example 1 In one embodiment, the compound of Example 1

[00086] is provided as a crystalline material comprising one or more crystalline forms. Examples of suitable crystalline forms of the compound of Example 1 include Forms N-1, A-2, and EA-3.
[00087] In one embodiment, the compound of Example 1 is provided as a crystalline material comprising the first crystalline form. A first crystalline form of the compound of Example 1 comprises a liquid crystalline form referred to herein as "Form N-1" or "Form N-1".
[00088] In one embodiment, Form N-1 of the compound of Example 1 is characterized by unit cell parameters approximately equal to the following: Cell dimensions: a = 9.41 A b = 17.74 A c = 31.94 A α = 90.0° β = 98.4° Y = 90.0° Space group: P21 Example molecules 1/asymmetric unit: 4 Volume/Number of molecules in unit cell = 659 A3 Density (calculated) = 1.402 g /cm3,
[00089] wherein the unit cell parameters of Form N-1 are measured at a temperature of about -10°C.
[00090] In another embodiment, the N-1 Form of the compound of Example 1 is characterized by a simulated x-ray powder diffraction (PXRD) pattern substantially in accordance with the pattern shown in Figure 1 and/or by a pattern of PXRD observed substantially in accordance with the pattern shown in Figure 1.
[00091] In yet another embodiment, the N-1 Form of the compound of Example 1 is characterized by a PXRD pattern (CuKα δ =1.5418A at a temperature of about 25°C) comprising four or more, preferably five or more, 2 θ values selected from: 5.7±0.2, 7.5±0.2, 10.3±0.2, 10.7±0.2, 15.2±0.2, 16 .8±0.2, 20.2±0.2, and 20.7±0.2, wherein the Form N-1 PXRD standard is measured at a temperature of about 20°C.
[00092] Yet in yet another embodiment, Form N-1 of Example 1 is characterized by fractional atomic coordinates substantially as listed in Table 1. Table 1: Fractional Atomic Coordinates of Form N-1 of Example 1 Calculated at a Temperature of about 25°C; Atomic Coordinates (x104) and Equivalent Isotropic Displacement Parameters (A2x 103)




*U(eq) is defined as one-third of the trace of the orthogonalized tensor Uij.
[00093] However, even in yet another embodiment, the N-1 form of the compound of Example 1 is substantially pure.
[00094] However, in yet another embodiment, the N-1 form of the compound of Example 1 contains at least about 90% by weight, preferably at least about 95% by weight, and more preferably at least about of 99% by weight, based on the weight of Form N-1 of the compound of Example 1.
[00095] In yet another embodiment, a substantially pure Form N-1 of the compound of Example 1 has substantially pure phase homogeneity of less than about 10%, preferably less than about 5%, and more preferably less than about 2 % of the total peak area of the experimentally measured PXRD pattern arising from peaks that are absent from the simulated PXRD pattern. More preferably, the substantially pure crystalline Form N-1 has substantially pure phase homogeneity with less than about 1% of the total peak area of the experimentally measured PXRD pattern arising from peaks that are absent from the simulated PXRD pattern.
[00096] In another embodiment, the crystalline form of the compound of Example 1 consists essentially of Form N-1. The crystalline form of this embodiment may comprise at least about 90% by weight, preferably at least about 95% by weight, and more preferably at least about 99% by weight, based on the weight of the crystalline form, Form N -1 of the compound of Example 1.
[00097] In yet another embodiment, a pharmaceutical composition comprising Form N-1 of the compound of Example 1 is provided; and at least one pharmaceutically acceptable carrier and/or diluent.
[00098] In yet another embodiment, a pharmaceutical composition comprises substantially pure Form N-1 of the compound of Example 1; and at least one pharmaceutically acceptable carrier and/or diluent.
[00099] However, in yet another embodiment, a therapeutically effective amount of Form N-1 of the compound of Example 1 is combined with at least one pharmaceutically acceptable carrier and/or diluent to provide at least one pharmaceutical composition.
[000100] In one embodiment, the compound of Example 1 is provided in a second crystalline form. The second crystalline form is an acetone solvate crystalline form referred to herein as "Form A-2" or "Form A-2". Form A-2 comprises about one molecule of acetone for each molecule of Example 1.
[000101] In one embodiment, the Form of A-2 is characterized by unit cell parameters approximately equal to the following: Cell dimensions: a = 9.25 A b = 17.11 A c = 19.63 A α = 90 .0° β = 99.2° Y = 90.0° Space group: P21 Example molecules 1/asymmetric unit: 2 Volume/number of molecules in unit cell = 767 A3 Density (calculated) = 1.331 g/cm3,
[000102] wherein the unit cell parameters of Form A-2 are measured at a temperature of about -50°C.
[000103] In another embodiment, Form A-2 is characterized by a simulated x-ray powder diffraction (PXRD) pattern substantially in accordance with the pattern shown in Figure 2 and/or by an observed PXRD pattern substantially in accordance with according to the pattern shown in Figure 2.
[000104] Yet in yet another embodiment, Form A-2 of Example 1 is characterized by fractional atomic coordinates substantially as listed in Table 2. Table 2: Fractional Atomic Coordinates of Form A-2 of Example 1 Calculated at a Temperature of about 25°C; Atomic Coordinates (x104) and Equivalent Isotropic Displacement Parameters (A2 x 103)


*U(eq) is defined as one-third of the trace of the orthogonalized tensor Uij.
[000105] However, even in yet another embodiment, the A-2 form of the compound of Example 1 is substantially pure.
[000106] However, in yet another embodiment, the A-2 form of the compound of Example 1 contains at least about 90% by weight, preferably at least about 95% by weight, and more preferably at least about 95% by weight. 99% by weight, based on weight of the second crystalline form, Form A-2.
[000107] In yet another embodiment, a substantially pure second crystalline form has substantially pure phase homogeneity with less than about 10%, preferably less than about 5%, and more preferably less than about 2% of the total peak area of the experimentally measured PXRD pattern arising from peaks that are absent from the simulated PXRD pattern. Preferably, a substantially pure second crystalline form has substantially pure phase homogeneity with less than about 1% of the total peak area of the experimentally measured PXRD pattern arising from peaks that are absent from the simulated PXRD pattern.
[000108] In another embodiment, the second crystalline form of the compound of Example 1 consists essentially of Form A-2. The second crystalline form of this embodiment can comprise at least about 90% by weight, preferably at least about 95% by weight, and more preferably at least about 99% by weight, based on the weight of the second crystalline form, Form A -two.
[000109] In one embodiment, the compound of Example 1 is provided in a third crystalline form. The third crystalline form is an ethyl acetate solvate crystalline form referred to herein as "Form EA-3" or "Form EA-3". Form EA-3 comprises about one molecule of ethyl acetate for each molecule of Example 1.
[000110] In one embodiment, the EA-3 Form is characterized by unit cell parameters approximately equal to the following: Cell dimensions: a = 8.84 A b = 15.95 A c = 22.38 A α = 90 .0° β = 90.0° Y = 90.0° Space group: P212121 Example molecules 1/asymmetric unit: 1 Volume/number of molecules in unit cell = 789 Â3 Density (calculated) = 1.357 g/cm3,
[000111] wherein the unit cell parameters of Form EA-3 are measured at a temperature of about -50°C.
[000112] In another embodiment, Form EA-3 is characterized by a simulated x-ray powder diffraction (PXRD) pattern substantially in accordance with the pattern shown in Figure 3 and/or by an observed PXRD pattern substantially from according to the pattern shown in Figure 3.
[000113] In yet another embodiment, the EA-3 Form of the compound of Example 1 is characterized by a PXRD pattern (CuKaÀ= 1.5418Â at a temperature of about 25°C) comprising four or more, preferably five or more, 2θ values selected from: 6.8±0.2, 9.6±0.2, 10.6±0.2, 15.4±0.2, 20.5±0.2, 21.0 ±0.2, and 24.8±0.2, where the Form N-1 PXRD standard is measured at a temperature of about 20°C.
[000114] Yet in yet another embodiment, Form EA-3 of Example 1 is characterized by fractional atomic coordinates substantially as listed in Table 3. Table 3: Fractional Atomic Coordinates of Form EA-3 of Example 1 Calculated at a Temperature of about 25°C; Atomic Coordinates (x104) and Equivalent Isotropic Displacement Parameters (A2x 103)


*U(eq) is defined as one-third of the trace of the orthogonalized tensor Uij.
[000115] However, even in yet another embodiment, Form EA-3 of the compound of Example 1 is substantially pure.
[000116] However, in yet another embodiment, the EA-3 Form of the compound of Example 1 contains at least about 90% by weight, preferably at least about 95% by weight, and more preferably at least about 99% by weight, based on the weight of the third crystalline form, Form EA-3.
[000117] In yet another embodiment, a substantially pure Form EA-3 has substantially pure phase homogeneity with less than about 10%, preferably less than about 5%, and more preferably less than about 2% peak area total experimentally measured PXRD pattern arising from peaks that are absent from the simulated PXRD pattern. Even more preferably, the substantially crystalline Form EA-3 has substantially pure phase homogeneity with less than about 1% of the total peak area of the experimentally measured PXRD pattern arising from peaks that are absent from the simulated PXRD pattern.
[000118] In another embodiment, the third crystalline form of the compound of Example 1 consists essentially of Form EA-3. The third crystalline form of this embodiment can comprise at least about 90% by weight, preferably at least about 95% by weight, and more preferably at least about 99% by weight, based on the weight of the third crystalline form, Form EA -3.
[000119] In one embodiment, the compound of Example 1 is provided in a fourth crystalline form. The fourth crystalline form is a tetrahydrofuran solvate crystalline form referred to herein as "Form THF-2" or "Form THF-2". Form THF-2 comprises about one molecule of tetrahydrofuran for each molecule of Example 1.
[000120] In one embodiment, the THF-2 Form is characterized by unit cell parameters approximately equal to the following: Cell dimensions: a = 9.34 A b = 16.44 A c = 20.60 A α = 90 .0° β = 102.8° Y = 90.0° Space group: P21 Example 1 molecules/asymmetric unit: 2 Tetrahydrofuran molecules/asymmetric unit: 2 Volume = 3082 A3,
[000121] wherein the unit cell parameters of Form THF-2 are measured at a temperature of about -50°C.
[000122] In another embodiment, the THF-2 Form is characterized by a simulated x-ray powder diffraction (PXRD) pattern substantially in accordance with the pattern shown in Figure 4 and/or by an observed PXRD pattern substantially from according to the pattern shown in Figure 4.
[000123] In yet another embodiment, the THF-2 Form of the compound of Example 1 is characterized by a PXRD pattern (CuKaÀ = 1.5418Â at a temperature of about 25°C) comprising four or more, preferably five or more, 2θ values selected from: 6.9±0.2, 9.6±0.2, 11.2±0.2, 12.6±0.2, 16.6±0.2, 21 .4±0.2, and 24.2±0.2, wherein the Form N-1 PXRD standard is measured at a temperature of about 20°C.
[000124] Yet in yet another embodiment, the THF-2 Form of Example 1 is characterized by fractional atomic coordinates substantially as listed in Table 4. Table 4: Fractional Atomic Coordinates of the THF-2 Form of Example 1 (Not Including Molecules of Solvent) Calculated at a Temperature of about -50°C; Atomic Coordinates (x104) and Equivalent Isotropic Displacement Parameters (A2x 103)


* U(eq) is defined as one third of the trace of the orthogonalized tensor Uij.
[000125] However, even in yet another embodiment, the THF-2 form of the compound of Example 1 is substantially pure.
[000126] However, in yet another embodiment, the THF-2 form of the compound of Example 1 contains at least about 90% by weight, preferably at least about 95% by weight, and more preferably at least about 99 % by weight, based on the weight of the fourth crystalline form, Form THF-2.
[000127] In yet another embodiment, a substantially pure Form THF-2 has substantially pure phase homogeneity of less than about 10%, preferably less than about 5%, and more preferably less than about 2% area of the total peak of the experimentally measured PXRD pattern arising from peaks that are absent from the simulated PXRD pattern. Even more preferably, the substantially crystalline Form THF-2 has substantially pure phase homogeneity with less than about 1% of the total peak area of the experimentally measured PXRD pattern arising from peaks that are absent from the simulated PXRD pattern.
[000128] In another embodiment, the fourth crystalline form of the compound of Example 1 consists essentially of Form THF-2. The fourth crystalline form of this embodiment may comprise at least about 90% by weight, preferably at least about 95% by weight, and more preferably at least about 99% by weight, based on the weight of the fourth crystalline form, THF Form -two. Crystalline Form of Compound of Example 2 In one embodiment, the compound of Example 2

[000129] is provided as a crystalline material comprising a crystalline form. An example of a suitable crystalline form of the compound of Example 2 is Form M2-1. Form M2-1 comprises approximately two molecules of methanol for each molecule of Example 2.
[000130] In one embodiment, Form M2-1 of the compound of Example 2 is characterized by unit cell parameters approximately equal to the following: Cell dimensions: a = 8.44 Å b = 21.02 Å c = 17.52 Å α = 90.0° β = 90.88° Y = 90.0° Space group: P21 Example molecules 2/asymmetric unit: 2 Volume/Number of molecules in unit cell = 777 Å 3 Density (calculated ) = 1.297 g/cm3,
[000131] wherein the unit cell parameters of Form M-1 are measured at a temperature of about -100°C.
[000132] In another embodiment, the Form of M2-1 is characterized by a simulated x-ray powder diffraction (PXRD) pattern substantially in accordance with the pattern shown in Figure 5 and/or by an observed PXRD pattern substantially from according to the pattern shown in Figure 5.
[000133] In yet another embodiment, Form M2-1 of the compound of Example 2 is characterized by a PXRD pattern (CuKaÀ =1.5418A at a temperature of about 25°C) comprising four or more, preferably five or more, 2θ values selected from: 8.2±0.2, 12.2±0.2, 14.2±0.2, 15.1±0.2, 16.8±0.2, 17.3 ±0.2, and 23.0±0.2, where the Form M2-1 PXRD standard is measured at a temperature of about 20°C.
[000134] Yet in yet another embodiment, Form M2-1 of Example 2 is characterized by fractional atomic coordinates substantially as listed in Table 5. Table 5: Fractional Atomic Coordinates of Form M2-1 Calculated at a Temperature of About 25°C; Atomic Coordinates (x 104) and Equivalent Isotropic Displacement Parameters (A2x 103) eg 2, Form M2-1



[000135] U(eq) is defined as one third of the trace of the orthogonalized tensor Uij.
[000136] However, even in yet another embodiment, the M2-1 form of the compound of Example 2 is substantially pure.
[000137] However, in yet another embodiment, the M2-1 form of the compound of Example 2 contains at least about 90% by weight, preferably at least about 95% by weight, and more preferably at least about 99% by weight, based on the weight of the crystalline form, Form M2-1.
[000138] In yet another embodiment, a substantially pure crystalline form of Form M2-1 has substantially pure phase homogeneity of less than about 10%, preferably less than about 5%, and more preferably less than about 5%. 2% of the total peak area of the experimentally measured PXRD pattern arising from peaks that are absent from the simulated PXRD pattern. Even more preferably, the substantially pure crystalline form of M2-1 has substantially pure phase homogeneity with less than about 1% of the total peak area of the experimentally measured PXRD pattern arising from peaks that are absent from the simulated PXRD pattern.
[000139] In another embodiment, the crystalline form of the compound of Example 2 consists essentially of Form M2-1. The crystalline form of this embodiment may comprise at least about 90% by weight, preferably at least about 95% by weight, and more preferably at least about 99% by weight, based on the weight of the crystalline form, Form M2 -1.
[000140] Compounds according to Formula (I) may be administered by any means suitable for the condition being treated which may depend on the need for site-specific treatment or amount of compound of Formula (I) to be delivered.
[000141] Also encompassed within this invention is a class of pharmaceutical compositions comprising the compound of Formula (I) or prodrug thereof; and one or more pharmaceutically acceptable, non-toxic carriers and/or diluents and/or adjuvants (collectively referred to herein as "carrier" materials) and, if desired, other active ingredients. The compounds of Formula (I) may be administered by any suitable route, preferably in the form of a pharmaceutical composition adapted to such route, and at a dose effective for the intended treatment. The compounds and compositions of the present invention may, for example, be administered orally, mucosally, or parenterally including intravascularly, intravenously, intraperitoneally, subcutaneously, intramuscularly, and intrasternally in unit dosage formulations that contain the pharmaceutically carriers, adjuvants and vehicles. conventional acceptable. For example, the pharmaceutical carrier may contain a mixture of mannitol or lactose and microcrystalline cellulose. The mixture may contain additional components such as a lubricating agent, for example magnesium stearate and a disintegrating agent such as crospovidone. The carrier mixture can be filled into a gelatin capsule or compressed into a tablet. The pharmaceutical composition can be administered as an oral dosage form or an infusion, for example.
[000142] For oral administration, the pharmaceutical composition may be in the form of, for example, a tablet, capsule, suspension, or liquid. The pharmaceutical composition is preferably made in the form of a dosage unit which contains a particular amount of the active ingredient. For example, the pharmaceutical composition may be provided as a tablet or capsule comprising an amount of active ingredient in the range of from about 1 to 2000 mg, preferably from about 1 to 500 mg, and more preferably from about 5 to 150 mg. mg. An adequate daily dose for a human or other mammal may vary, depending largely on the patient's condition and other factors, but may be determined using routine methods.
[000143] Any pharmaceutical composition contemplated herein may, for example, be delivered orally by means of any suitable and acceptable oral preparations. Exemplary oral preparations include, but are not limited to, for example, tablets, troches, lozenges, aqueous and oily suspensions, dispersible powders or granules, emulsions, hard and soft capsules, syrups, and elixirs. Pharmaceutical compositions intended for oral administration may be prepared according to any method known in the art to manufacture pharmaceutical compositions intended for oral administration. To provide pharmaceutically palatable preparations, a pharmaceutical composition according to the invention may contain at least one agent selected from sweetening agents, flavoring agents, coloring agents, demulcents, antioxidants, and preserving agents.
[000144] A tablet may, for example, be prepared by administering at least one compound of Formula (I) with at least one non-toxic pharmaceutically acceptable excipient suitable for the manufacture of tablets. Exemplary excipients include, but are not limited to, for example, inert diluents, such as, for example, calcium carbonate, sodium carbonate, lactose, calcium phosphate, and sodium phosphate; granulating and disintegrating agents, such as, for example, microcrystalline cellulose, croscarmellose sodium, corn starch, and alginic acid; binding agents, such as, for example, starch, gelatin, polyvinyl pyrrolidone, and acacia; and lubricating agents, such as, for example, magnesium stearate, stearic acid, and talc. Additionally, a tablet may be uncoated, or coated by known techniques to mask the bad taste of an unpleasant tasting drug, or delay the disintegration and absorption of the active ingredient in the gastrointestinal tract thereby sustaining the effects of the active ingredient for a longer period. long. Exemplary water-soluble taste masking materials include, but are not limited to, hydroxypropylmethylcellulose and hydroxypropylcellulose. Exemplary time delay materials include, but are not limited to, ethyl cellulose and cellulose acetate butyrate.
[000145] Hard gelatine capsules can, for example, be prepared by mixing at least one compound of Formula (I) with at least one inert solid diluent, such as, for example, calcium carbonate; Calcium phosphate; and kaolin.
[000146] Soft gelatine capsules can, for example, be prepared by mixing at least one compound of Formula (I) with at least one water-soluble carrier, such as, for example, polyethylene glycol; and at least one oily medium, such as, for example, peanut oil, liquid paraffin, and olive oil.
[000147] An aqueous suspension can be prepared, for example, by mixing at least one compound of Formula (I) with at least one suitable excipient for the manufacture of an aqueous suspension. Exemplary excipients suitable for making an aqueous suspension include, but are not limited to, for example, suspending agents such as, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, alginic acid, polyvinylpyrrolidone, gum tragacanth, and acacia gum; dispersing or wetting agents, such as, for example, a naturally occurring phosphatide, for example, lecithin; condensation products of alkylene oxide with fatty acids, such as, for example, polyoxyethylene stearate; condensation products of ethylene oxide with long-chain aliphatic alcohols, such as, for example, heptadecaethylene-oxycetanol; condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol, such as, for example, polyoxyethylene sorbitol monooleate; and condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as, for example, polyethylene sorbitan monooleate. An aqueous suspension may likewise contain at least one preservative, such as, for example, n-propyl ethyl p-hydroxybenzoate; at least one coloring agent; at least one flavoring agent; and/or at least one sweetening agent, including but not limited to, for example, sucrose, saccharin and aspartame.
[000148] Oil suspensions can, for example, be prepared by suspending at least one compound of Formula (I) in a vegetable oil, such as, for example, peanut oil; olive oil; Sesame oil; and coconut oil; or in mineral oil, such as, for example, liquid paraffin. An oily suspension may likewise contain at least one thickening agent, such as, for example, beeswax; hard paraffin; and cetyl alcohol. To provide a flavorful oily suspension, at least one of the sweetening agents already described hereinbefore, and/or at least one flavoring agent may be added to the oily suspension. An oily suspension may also contain at least one preservative, including, but not limited to, for example an antioxidant, such as, for example, butylated hydroxyanisole and alpha-tocopherol.
[000149] Dispersible powders and granules can, for example, be prepared by mixing at least one compound of Formula (I) with at least one dispersing and/or wetting agent; at least one suspending agent; and/or at least one preservative. Suitable dispersing agents, wetting agents and suspending agents are as already described above. Exemplary preservatives include, but are not limited to, for example antioxidants, for example ascorbic acid. In addition, dispersible powders and granules can likewise contain at least one excipient, including, but not limited to, for example sweetening agents; flavoring agents; and coloring agents.
[000150] An emulsion of at least one compound of Formula (I) may, for example, be prepared as an oil-in-water emulsion. The oil phase of the emulsions comprising the compounds of Formula (I) may comprise known ingredients in a known manner. The oil phase may be provided by, but is not limited to, for example a vegetable oil, such as, for example, olive oil and peanut oil; a mineral oil, such as, for example, liquid paraffin; and mixtures thereof. While the phase may comprise only one emulsifier, it may comprise a mixture of at least one emulsifier with a fat or an oil or with a fat and an oil. Suitable emulsifying agents include, but are not limited to, for example naturally occurring phosphatides, for example soy lecithin; esters or partial esters derived from fatty acids and hexitol anhydrides, such as, for example, sorbitan monooleate; and condensation products of partial esters with ethylene oxide, such as, for example, polyoxyethylene sorbitan monooleate. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabilizer. It is likewise preferred to include an oil and a fat equally. Together, the emulsifier(s) with or without stabilizer(s) prepare(s) the so-called emulsifying wax, and the wax together with the oil and fat prepare the so-called emulsifying ointment base which forms the dispersed phase. oil from cream formulations. An emulsion may likewise contain a sweetening agent, flavoring agent, a preservative, and/or an antioxidant. Emulsifiers and emulsion stabilizers suitable for use in the formulation of the present invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate, glyceryl distearate alone or with a wax, or other materials as well. known in the art.
[000151] The compounds of Formula (I) may, for example, also be delivered intravenously, subcutaneously and/or intramuscularly by any suitable and pharmaceutically acceptable injectable form. Exemplary injectable forms include, but are not limited to, for example, sterile aqueous solutions comprising acceptable vehicles and solvents, such as, for example, water, Ringer's solution, and isotonic sodium chloride solution; sterile oil-in-water microemulsions; and aqueous or oleaginous suspensions.
[000152] Formulations for parenteral administration may be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions and suspensions may be prepared from sterile powders or granules using one or more of the vehicles or diluents mentioned for use in formulations for oral administration or using other suitable dispersing or wetting agents and suspending agents. The compounds can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, gum tragacanth, and/or various buffers. Other adjuvants and modes of administration are well and widely known in the pharmaceutical art. The active ingredient can be similarly administered by injection as a composition with suitable vehicles including saline, dextrose, or water, or with cyclodextrin (i.e. CAPTISOL®), cosolvent solubilization (i.e. propylene glycol) or micellar solubilization. (i.e. Tween 80).
[000153] The sterile injectable preparation may likewise be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.
[000154] A sterile injectable oil-in-water microemulsion can, for example, be prepared by 1) dissolving at least one compound of Formula (I) in an oil phase, such as, for example, an oil mixture of soy and lecithin; 2) combining Formula (I) containing the oil phase with a water and glycerol mixture; and 3) processing the blend to form a microemulsion.
[000155] A sterile aqueous or oleaginous suspension can already be prepared according to methods known in the art. For example, a sterile aqueous solution or suspension can be prepared with a non-toxic parenterally acceptable diluent or solvent, such as, for example, 1,3-butane diol; and a sterile oleaginous suspension may be prepared with a sterile non-toxic acceptable solvent or suspending medium, such as, for example, sterile fixed oils, for example, synthetic mono- or diglycerides; and fatty acids, such as, for example, oleic acid.
[000156] Pharmaceutically acceptable carriers, adjuvants and vehicles that can be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, aluminum, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS). ) such as d-alpha-tocopheryl polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens, polyethoxylated castor oil such as CREMOPHOR surfactant (BASF), or other similar polymeric release matrices, whey proteins such as such as human serum albumin, buffer substances such as phosphate, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate , sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, substances with m based on cellulose, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, block polymers of polyethylene-polyoxypropylene, polyethylene glycol and lanolin. Cyclodextrins such as alpha, beta, and gamma-cyclodextrins, or chemically modified derivatives such as hydroxyalkylcyclodextrins, including 2- and 3-hydroxypropyl-cyclodextrins, or other solubilized derivatives may also be advantageously used to enhance the release of compounds of the described formulas. on here.
[000157] The pharmaceutically active compounds of this invention can be processed according to conventional methods of pharmacy to produce medicinal agents for administration to patients, including humans and other mammals. Pharmaceutical compositions may undergo conventional pharmaceutical operations such as sterilization and/or may contain pharmaceutical adjuvants such as preservatives, stabilizers, wetting agents, emulsifiers, buffers, etc. Tablets and pills can additionally be prepared with enteric coatings. Such compositions may also comprise adjuvants, such as wetting, sweetening, flavoring and perfuming agents.
[000158] The amounts of compounds that are administered and the dosage regimen for treating a disease condition with the compounds and/or compositions of this invention depend on a variety of factors, including age, weight, sex, medical condition. the individual, the type of disease, the severity of the disease, the routine and frequency of administration, and the particular compound employed. Thus, the dosing regimen can vary widely, but can be routinely determined using standard methods. A daily dose of from about 0.001 to 100 mg/kg of body weight, preferably between about 0.005 and about 50 mg/kg of body weight and more preferably between about 0.01 to 10 mg/kg of body weight, can be appropriate. The daily dose can be administered in one to four doses per day.
[000159] For therapeutic purposes, the active compounds of this invention are normally combined with one or more adjuvants appropriate to the indicated route of administration. If administered orally, the compounds can be mixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acids, alkyl cellulose esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of acids. phosphoric and sulfuric acid, gelatin, acacia gum, sodium alginate, polyvinylpyrrolidone, and/or polyvinyl alcohol, and then converted into tablets or capsules for convenient administration. Such capsules or tablets may contain a controlled release formulation as may be provided in a dispersion of the active compound in hydroxypropylmethyl cellulose.
[000160] The pharmaceutical compositions of this invention comprise the compound of formula (I), or a prodrug thereof, and optionally an additional agent selected from any pharmaceutically acceptable carrier, adjuvant, and vehicle. Alternative compositions of this invention comprise a compound of formula (I) described herein, or a prodrug thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. UTILITY
[000161] The compounds of formula (I) are useful for the treatment of cancer, for example, Notch activation dependent cancers. Notch activation has been implicated in the pathogenesis of several solid tumors including ovarian, pancreatic, as well as breast and hematologic tumors such as leukemias, lymphomas, and multiple myeloma.
[000162] In one embodiment, a method provided for treating cancer comprises administering to a mammal in need thereof a compound of formula (I) or a prodrug thereof. The method of this modality can be used to treat a variety of cancers, including but not limited to bladder cancer, breast cancer, colorectal cancer, gastric cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer including non-small cell lung cancer (NSCLC), ovarian cancer, pancreatic cancer, gallbladder cancer, prostate cancer, thyroid cancer, osteosarcoma, rhabdomyosarcoma, malignant fibrous histiocytoma (MFH), fibrosarcoma, glioblastomas /astrocytomas, neuroblastoma, melanoma, T-cell acute lymphoblastic leukemia (T-ALL), and mesothelioma. For example, the method of this modality is used to treat breast cancer, colon cancer, or pancreatic cancer. Preferably, the mammal is a human. For example, a therapeutically effective amount for treating cancer can be administered in the method of the present embodiment. The method of this embodiment includes administering the compound having the structure:

[000163] The method of this embodiment also includes the administration of the compound having the structure:

[000164] Routes of administration in the present embodiment include parenteral administration and oral administration.
[000165] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof, wherein said cancer is colorectal cancer. Preferably, the mammal is a human. For example, a therapeutically effective amount for treating cancer can be administered in the method of the present embodiment. Routes of administration in the present embodiment include parenteral administration and oral administration.
[000166] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof, wherein said cancer is triple negative breast cancer. Preferably, the mammal is a human. For example, a therapeutically effective amount for treating cancer can be administered in the method of the present embodiment. Routes of administration in the present embodiment include parenteral administration and oral administration.
[000167] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof, wherein said cancer is non-cellular lung cancer. little. Preferably, the mammal is a human. For example, a therapeutically effective amount for treating cancer can be administered in the method of the present embodiment. Routes of administration in the present embodiment include parenteral administration and oral administration.
[000168] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof, wherein said cancer is pancreatic cancer. Preferably, the mammal is a human. For example, a therapeutically effective amount for treating cancer can be administered in the method of the present embodiment. Routes of administration in the present embodiment include parenteral administration and oral administration.
[000169] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof, wherein said cancer is ovarian cancer. Preferably, the mammal is a human. For example, a therapeutically effective amount for treating cancer can be administered in the method of the present embodiment. Routes of administration in the present embodiment include parenteral administration and oral administration.
[000170] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof, wherein said cancer is melanoma. Preferably, the mammal is a human. For example, a therapeutically effective amount for treating cancer can be administered in the method of the present embodiment. Routes of administration in the present embodiment include parenteral administration and oral administration.
[000171] In one embodiment, the use of a compound of formula (I) or a prodrug thereof, in the manufacture of a medicament for the treatment of cancer is provided. Preferably, in the present embodiment, cancers undergoing treatment include one or more of bladder cancer, breast cancer, colorectal cancer, gastric cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer including non-small cell lung disease (NSCLC), ovarian cancer, pancreatic cancer, gallbladder cancer, prostate cancer, thyroid cancer, osteosarcoma, rhabdomyosarcoma, malignant fibrous histiocytoma (MFH), fibrosarcoma, glioblastomas/astrocytomas, neuroblastoma, melanoma, T-cell acute lymphoblastic leukemia (T-ALL), and mesothelioma. Suitable medicaments of the present embodiment include medicaments for parenteral administration, such as, for example, solutions and suspensions, and medicaments for oral administration, such as, for example, tablets, capsules, solutions, and suspensions.
[000172] One embodiment provides a compound of formula (I) or a prodrug thereof, for use in therapy in the treatment of cancer. In the present embodiment, cancers undergoing treatment include one or more of bladder cancer, breast cancer, colorectal cancer, gastric cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer including non-small cell (NSCLC), ovarian cancer, pancreatic cancer, gallbladder cancer, prostate cancer, thyroid cancer, osteosarcoma, rhabdomyosarcoma, malignant fibrous histiocytoma (MFH), fibrosarcoma, glioblastomas/astrocytomas, neuroblastoma, melanoma , T-cell acute lymphoblastic leukemia (T-ALL), and mesothelioma.
[000173] In one embodiment, a method is provided for treating cancer in a mammal in which the cancer is dependent on Notch activation, comprising administering to the patient a compound of formula (I) or a prodrug thereof. The method of this modality can be used to treat a variety of cancers, including but not limited to bladder cancer, breast cancer, colorectal cancer, gastric cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer including non-small cell lung cancer (NSCLC), ovarian cancer, pancreatic cancer, gallbladder cancer, prostate cancer, thyroid cancer, osteosarcoma, rhabdomyosarcoma, malignant fibrous histiocytoma (MFH), fibrosarcoma, glioblastomas /astrocytomas, neuroblastoma, melanoma, T-cell acute lymphoblastic leukemia (T-ALL), and mesothelioma. Preferably, the method of this embodiment is used to treat breast cancer, colon cancer, or pancreatic cancer. Preferably, the mammal is a human. For example, a therapeutically effective amount for treating cancer can be administered in the method of the present embodiment. Suitable routes of administration include parenteral administration and oral administration.
[000174] In the treatment of cancer, a combination of chemotherapeutic agents and/or other treatments (eg, radiation therapy) is often advantageous. The second (or third) agent may have the same or different mechanism of action than the primary therapeutic agent. For example, drug combinations may be employed, where two or more drugs being administered act in different ways or at different stages of the cell cycle, and/or where two or more drugs have non-overlapping toxicities or side effects, and/or wherein the drugs being combined with each other have demonstrated efficacy in treating the particular disease state manifested by the patient.
[000175] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof; and administering one or more additional anticancer agents.
[000176] The phrase "additional anticancer agent" refers to a drug selected from any one or more of the following: alkylating agents (including nitrogen mustards, alkyl sulfonates, nitrosoureas, ethylenimine derivatives, and triazenes); antiangiogenics (including matrix metalloproteinase inhibitors); antimetabolites (including adenosine deaminase inhibitors, folic acid antagonists, purine analogues, and pyrimidine analogues); antibiotics and antibodies (including monoclonal antibodies, CTLA-4 antibodies, anthracyclines); aromatase inhibitors; cell cycle response modifiers; farnesyl protein transferase inhibitors; hormonal and anti-hormonal agents, and steroids (including synthetic analogues, glucocorticoids, estrogens/antiestrogens [e.g., SERMs], erogens/antiandrogens, progestins, progesterone receptor agonists, and hormone-releasing agonists and antagonists). luteinizing hormone [LHRH]); insulin-like growth factor (IGF)/insulin-like growth factor receptor (IGFR) system modulators (including IGFR1 inhibitor); integrin signaling inhibitors; kinase inhibitors (including multi-kinase inhibitors and/or Src kinase or Src/abl inhibitors, cyclin-dependent kinase [CDK] inhibitors, panHer, Her-1 and Her-2 antibodies, VEGF inhibitors, including anti- -VEGF, EGFR inhibitors, mitogen-activated protein [MAP] inhibitors, MET inhibitors, MEK inhibitors, Aurora kinase inhibitors, PDGF inhibitors, and other tyrosine kinase inhibitors or serine/threonine kinase inhibitors; microtubule disrupting agents such as ecteinascidins or their analogues and derivatives; microtubule stabilizing agents such as taxanes, and the naturally occurring epothilones and their synthetic and semi-synthetic analogues; destabilizing agents, microtubule binding (including vinca alkaloids) ; topoisomerase inhibitors; prenyl protein transferase inhibitors; platinum coordination complexes; signal transduction inhibitors; and other agents used as anticancer and cytotoxic agents such as such as biological response modifiers, growth factors, and immune modulators.
[000177] Accordingly, the compounds of the present invention may be administered in combination with other anti-cancer treatments useful in the treatment of cancer or other proliferative diseases. The invention herein also comprises the use of a compound of formula (I) or prodrug thereof in the preparation of medicaments for the treatment of cancer, and/or comprises packaging a compound of formula (I) herein together with instructions that the compounds are used in combination with other anti-cancer or cytotoxic agents and treatments for the treatment of cancer. The present invention also comprises combinations of a compound of formula (I) and one or more additional agents in kit form, for example where they are packaged together or placed in separate packages to be sold as a kit, or where they are are packaged to be formulated together.
[000178] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof; administer dasatinib; and optionally, one or more additional anti-cancer agents.
[000179] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof; administer paclitaxel; and optionally, one or more additional anti-cancer agents.
[000180] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof; administering Tamoxifen; and optionally, one or more additional anti-cancer agents.
[000181] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof; administering a glucocorticoid; and optionally, one or more additional anti-cancer agents. An example of a suitable glucocorticoid is dexamethasone.
[000182] In one embodiment, a method is provided for treating cancer comprising administering to an animal in need thereof a compound of formula (I) or a prodrug thereof; administering carboplatin; and optionally, one or more additional anti-cancer agents.
[000183] The compounds of the present invention can be formulated or co-administered with other therapeutic agents that are selected for their particular utility in controlling side effects associated with the aforementioned conditions. For example, compounds of the invention can be formulated with agents to prevent nausea, hypersensitivity and gastric irritation, such as antiemetics, and H1 and H2 antihistamines.
[000184] In one embodiment, pharmaceutical compositions are provided comprising a compound of formula (I) or prodrug thereof; one or more additional agents selected from a kinase inhibitory agent (small molecule, polypeptide, and antibody), an immunosuppressant, an anticancer agent, an antiviral agent, an anti-inflammatory agent, an antifungal agent, an antibiotic, or a compound of antivascular hyperproliferation; and any carrier, adjuvant or vehicle.
[000185] The above other therapeutic agents, when employed in combination with the compounds of the present invention, may be used, for example, in those amounts indicated in the Physician's Desk Reference (PDR) or as otherwise determined by one skilled in the art. . In the methods of the present invention, such other therapeutic agents may be administered prior to, simultaneously with, or after administration of the inventive compounds.
[000186] The specific dose level and frequency of dosing for any particular individual may be varied and generally depend on a variety of a variety of factors, including but not limited to, for example, the bioavailability of the specific compound of formula (I) ) in the form administered, metabolic stability and intensity of action of the specific compound of formula (I), species, body weight, general health, sex, diet of the individual, mode and time of administration, rate of excretion, drug combination, and severity of the particular condition. For example, a daily dose of about 0.001 to 100 mg/kg of body weight, preferably between about 0.005 and about 50 mg/kg of body weight and more preferably between about 0.01 to 10 mg/kg of body weight body, may be appropriate. The daily dose can be administered in one to four doses per day.
[000187] Administration may be continuous, that is, every day, or intermittently. The terms "intermittent" or "intermittently" as used herein, mean stopping and starting at any regular or irregular intervals. For example, intermittent administration includes administration one to six days a week; cycle administration (e.g., daily administration for two to eight consecutive weeks, followed by a rest period of no administration for up to one week); or administration on alternate days.
[000188] In one embodiment, the compound of formula (I) is administered continuously to a patient in need thereof, one or more times daily. For example, a therapeutically effective amount of the compound of formula (I) is administered to a patient in need thereof, one or more times daily for continuous days.
[000189] In one embodiment, the compound of formula (I) is administered intermittently to a patient in need thereof, one or more times daily. For example, a therapeutically effective amount of the compound of formula (I) is administered to a patient in need thereof, one or more times daily on an intermittent schedule.
[000190] In one embodiment, the compound of formula (I) is administered to a patient in need thereof, one or more times daily for continuous days followed by one or more days without administration. Preferably, a therapeutically effective amount of the compound of formula (I) is administered. Examples of continuous dosing with a drug vacation are cycles of: 7 days on treatment followed by 7 days off treatment; 14 days on treatment followed by 7 days off treatment; and 7 days off treatment followed by 14 days off treatment. An on/off cycle may be repeated multiple times as required to treat a patient.
[000191] In one embodiment, the compound of formula (I) is administered to a patient in need thereof, according to an intermittent dosing schedule. Intermittent dosing schedules are repeating schedules including days when the patient is administered the compound of formula (I) and days when the patient is not administered the compound of formula (I). Examples of intermittent dosing schedules are: dosing four days each week for three continuous weeks followed by one week without dosing, and repeating at a four-week interval; dosing five days each week for two continuous weeks followed by one week without dosing, and repeating at a three-week interval; and dosing four days each week for one week followed by two weeks without dosing, and repeating at a three-week interval. Preferably, a therapeutically effective amount of the compound of formula (I) is administered.
[000192] In one embodiment, the compound of formula (I) is administered on one day, followed by 6 days off, and repeated on a weekly schedule.
[000193] In one embodiment, the compound of formula (I) is administered on two consecutive days, followed by 5 days off, and repeated on a weekly schedule.
[000194] In one embodiment, the compound of formula (I) is administered on three consecutive days followed by four days off, and repeated on a weekly schedule.
[000195] In one embodiment, the compound of formula (I) is administered in one day, followed by 10 to 13 days off. PREPARATION METHODS
[000196] The compounds of the present invention can be prepared in a number of ways well known to one skilled in the art of organic synthesis. The compounds of the present invention can be synthesized using the methods described using the methods described below, together with synthetic methods known in the art of synthetic organic chemistry, or variations thereon as appreciated by those skilled in the art. Preferred methods include, but are not limited to, those described below. All references cited herein are hereby incorporated in their entirety by reference.
[000197] The compounds of this invention can be prepared using the reactions and techniques described in these sections. The reactions are carried out in solvents appropriate for the reagents and materials used and are suitable for the transformations being carried out. In addition, in the description of the synthetic methods described below, it should be understood that all the proposed reaction conditions, including the choice of solvent, reaction atmosphere, reaction temperature, duration of experiment and preparation procedures, are chosen to be the conditions standards for that reaction, which should be readily recognized by one skilled in the art. It should be understood by one skilled in the art of organic synthesis that the functionality present in various portions of the molecule must be compatible with the proposed reactants and reactions. Such restrictions on substituents that are compatible with the reaction conditions will be readily apparent to one skilled in the art and alternative methods should therefore be used. This will sometimes require judgment to modify the order of synthetic steps or to select one particular process scheme over another in order to obtain a desired compound of the invention. It will also be recognized that another major consideration in designing any synthetic route in this field is the judicious choice of protecting group from the reactive functional groups present in the compounds described in this invention. An authoritative report that describes the many alternatives for the trained physician is Greene and others (Protective Groups in Organic Synthesis, third edition, Wiley and Sons (1999)).
[000198] Compounds of formula (I) may be prepared by reference to the methods illustrated in the following schemes. As shown in this regard, the end product is a compound having the same structural formula as formula (I). It will be understood that any compound of formula (I) can be produced by the schemes by the proper selection of appropriately substituted reagents. Solvents, temperatures, pressures, and other reaction conditions can easily be selected by one skilled in the art. Starting materials are commercially available or readily prepared by one skilled in the art. Compound constituents are as defined here or elsewhere in the specification.
[000199] The synthesis of compounds of formula (I) can be done using the methods summarized in Schemes 1 to 5. Scheme 1

[000200] The benzodiazepinone preparation (iv) can be carried out in a multitude of methods known to one skilled in the art. For example, as shown in Scheme 1, a suitably substituted 2-aminobenzophenone (i) (e.g., from Walsh, DA, Synthesis, 677 (1980); and references cited therein, or other methods known to one skilled in the art) can be coupled to the protected glycine derivative (ii) (PG = protecting group, for example PG = CBz, see Katritzki, AR, J. Org. Chem., 55:2206-2214 (1990)), treated with a reagent such as ammonium and subjected to cyclization to provide the benzodiazepinone (iii), according to the procedure outlined in the literature (e.g., Sherrill, RG et al., J. Org. Chem., 60:730 (1995); or other routes known to one skilled in the art). The resulting racemic mixture can be separated (using procedures known to one skilled in the art) to acquire the individual enantiomers, or used as a racemate. Likewise, if R3=H, (iii) can, for example, be treated with a reagent such as MeI and a base such as K2CO3 in a solvent such as DMF to prepare R3=Me.
[000201] Step 2: Deprotection of (iii) can be carried out in various ways known to one skilled in the art. For example, with PG = CBz, Compound (iii) can be treated with a reagent such as HBr in a solvent such as AcOH. Compound (iv) can be used as a racemate. Alternatively, compound (iv) may be subjected to enantiomeric resolution using standard methods (e.g. preparative chiral chromatography). scheme 2

[000202] Step 1: The first step of Scheme 2 is carried out by converting compound (v) to ester (vii) using one of the multiple modes known to one skilled in the art, such as treatment with a substituted acetimidate such as as compound (vi) in the presence of a reagent such as boron trifluoride etherate at an appropriate temperature in a solvent such as THF.
[000203] Step 2: Acid (viii) can be converted to compound (ix) in multiple ways known to one skilled in the art. For example, treatment of acid (viii) with a reagent such as oxalyl chloride in a solvent such as DCM yields acid chloride (ix). Compound (ix) can be treated with an oxazolidinone (x) under standard conditions to produce compound (xi) (Evans, D.A. et al., J. Am. Chem Soc., 112:4011 (1990)).
[000204] Step 3: Compound (xi) can be converted to compound (xii) in multiple ways (Baran, P. et al., J. Am. Chem. Soc., 130(34):11546 (2008 )). For example, compound (vii) is treated with a base such as LDA in a solvent such as toluene, at a low temperature such as -78°C under an inert atmosphere such as N 2 . The resulting mixture is added to a solution of compound (xi) treated with lithium chloride and a base such as LDA in a solvent such as toluene under an inert atmosphere such as N 2 . To the resulting mixture of the enolates of compounds (vii) and (xi) is added a compound, such as bis(2-ethylhexanoyloxy)copper, at a low temperature such as -78°C under an inert atmosphere such as N2 and heated to room temperature. environment to provide the compound (xii).
[000205] Step 4: The conversion of compound (xii) to (xiii) can be accomplished by treating it with reagents such as hydrogen peroxide and lithium hydroxide at an appropriate temperature, using a solvent mixture such as THF/water. If necessary, diastereoisomers can be separated at this point by silica gel chromatography or preparative HPLC. Alternatively, the mixture may be subjected to epimerization conditions, for example, by treatment with LDA and diethylaluminum chloride followed by quenching with methanol or acetic acid to enrich the desired diastereoisomer.
[000206] Step 5: Compound (xiii) can be coupled with benzodiazepinone (iv) in the presence of a coupling reagent such as TBTU and a base such as TEA, in a solvent such as DMF to provide compound (xiv ).
[000207] Step 6: Treatment of compound (xiv) with an acid such as TFA at an appropriate temperature such as 0°C, in a solvent such as DCM provides compound (xv).
[000208] Step 7: The conversion of compound (xv) to compound (xvi) can be carried out by coupling compound (xv) with an appropriate amine source such as ammonium chloride, a carbodiimide such as EDC, HOBT and a base such as TEA in a solvent such as DMF. If necessary, the diastereoisomeric mixture can be separated using an appropriate separation technique, such as chiral preparative chromatography. Scheme 3

[000209] Step 1: The preparation of benzodiazepinone (iii) can be carried out in the same way by cross-coupling a benzodiazepinone (xvii) containing a halogen atom such as chlorine (X=Cl) and a protecting group (PG) such as Boc , with an appropriate coupling pair such as a boronic acid under conditions known to one skilled in the art. For example, coupling of the halogen-containing moiety with a boronic acid occurs in the presence of a catalyst such as tetracis(triphenylphosphine)palladium(0), a base such as sodium carbonate and a solvent such as DME under an inert atmosphere such as N2. Scheme 4

[000210] Step 1: The first step of Scheme 4 involves treating Compound (xvii) with carboxylic acid (xix) in the presence of a carbodiimide such as DCC, a base such as TEA, and a catalyst such as DMAP in a solvent such as DCM provides Compound (xx).
[000211] Step 2: Conversion of Compound (xx) to Compound (xxi) may be carried out by treatment with a reagent such as sodium cyanoborohydride in the presence of an acid such as HCl under atmospheric conditions which may be inert, for example , under N2.
[000212] Step 3: Conversion from Compound (xxi) to Compound (xxiii) can proceed via Compound (xxii) supporting an appropriate leaving group (LG). For example, treatment of Compound (xxi) with a base such as 2,6-lutidine and a reagent such as trifluoromethanesulfonic anhydride in a solvent such as DCM at a suitable temperature such as -78°C provides the Compound triflate (xxii). Compound (xxii) can now be subjected to cross-coupling reaction conditions to yield Compound (xxiii). For example, treatment of Compound (xxii) with a suitably substituted coupling pair, for example a boronic acid, in the presence of a catalyst such as tetracyl(triphenylphosphine)palladium(0), a base such as potassium phosphate in a solvent such as dioxane under atmospheric conditions which may be inert, for example under N 2 , gives Compound (xxiii).
[000213] Step 4: Conversion of Compound (xxiii) to Compound (xxiv) can be performed by standard procedures known to one skilled in the art. For example, treatment of Compound (xxiii) in the presence of a catalyst such as Pd/C in a solvent such as methanol yields Compound (xxiv).
[000214] Step 5: Compound (xxv) can be obtained by coupling Compound (xxiv) with Compound (iv). For example, the transformation can be carried out using a reagent such as AlMe3 in a solvent such as DCM under an inert atmosphere such as N2. In this example, the mixture of diastereoisomers obtained can be used as a mixture or can be separated by an appropriate method such as chiral chromatography.
[000215] Step 6: Compound (xxv) is oxidized using an oxidizing agent such as Jones reagent, in a solvent such as acetone to produce Compound (xxvi). If the compound is then a diastereoisomeric mixture, then it can be used as a mixture or it can be separated using an appropriate method such as chiral chromatography.
[000216] Step 7: Conversion of Compound (xxvi) to Compound (xxvii) can be performed by standard procedures known to one skilled in the art. For example, coupling Compound (xxvi) with an appropriate amine source such as ammonium chloride, a carbodiimide such as EDC, HOBT and a base such as TEA in a solvent such as DMF provides Compound (xxvii). In this example, the compound can be enantiopure or if necessary the diastereoisomeric mixture can be separated using an appropriate separation technique, such as chiral chromatography. Scheme 5

[000217] Step 1: The first step of Scheme 5 is carried out by treating Compound (xxviii) with a reagent such as sodium nitrite in an acid such as H2SO4 and a solvent such as water to provide Compound xxix.
[000218] Step 2: Acid (xxix) is converted to compound (xxx) (PG = protecting group). For example, acid (xxix) is treated with an alcohol such as benzyl alcohol in a solvent such as toluene and an acid such as H2 SO4 to provide Compound xxx.
[000219] Step 3: Compound (xxxi) bearing a suitable leaving group can be prepared by treating Compound (xxx) with a base such as 2,6-lutidine and a reagent such as trifluoromethanesulfonic anhydride in a solvent such as DCM at an appropriate temperature.
[000220] Step 4: Compound (xxxii) can be converted to Compound (xxxiv) in multiple ways known to one skilled in the art. For example, treatment of acid chloride (xxxii), prepared from the corresponding carboxylic acid with a reagent such as oxalyl chloride in a solvent such as DCM, or obtained commercially, can be treated with an oxazolidinone (xxxiii) under standard conditions to produce Compound (xxxiv) (Evans, DA et al., J. Am. Chem Soc., 112:4011 (1990)).
[000221] Step 5: The preparation of Compound (xxxv) can be carried out by treating Compound (xxxiv) with a base such as LiHMDS in a solvent such as THF at an appropriate temperature such as -78°C and the resulting mixture Compound (xxxi) in a solvent such as THF is added.
[000222] Step 6: The Compound protecting group (xxxv) can be removed by many methods known to one skilled in the art. For example, a benzyl group can be removed by subjecting it to hydrogenation conditions using a palladium catalyst such as Pearlman's Catalyst in a solvent such as methanol to provide Compound (xxxvi).
[000223] Step 7: Compound (iv) is coupled with Compound (xxxvi) in the presence of a coupling reagent such as TBTU and a base such as TEA in a solvent such as DMF to provide Compound (xxxvii). If necessary, diastereoisomers can be separated using an appropriate method such as chiral preparative chromatography.
[000224] Step 8: Hydrolysis of Compound (xxxvii) can be carried out by treating it with hydrogen peroxide and lithium hydroxide at an appropriate temperature using a solvent mixture such as THF/water to produce Compound (xv). If necessary, diastereoisomers can be separated using an appropriate method such as preparative chiral chromatography. Scheme 6

[000225] Compound (xiii) in Scheme 2 can be prepared from compound (xi) by the synthetic sequence outlined in Scheme 6.
[000226] Step 1: The first step of Scheme 6 is carried out by treating Compound (xi) with a base such as sodium bis(trimethylsilyl)amide in a solvent such as THF at a low temperature such as -78°C under an inert atmosphere. The resulting enolate of (xi) is treated with a reagent such as tert-butyl bromoacetate to provide compound (xxxviii).
[000227] Step 2: The conversion of compound (xxxviii) to (xxxix) can be accomplished by treating compound (xxxviii) with reagents such as hydrogen peroxide and lithium hydroxide at an appropriate temperature using a solvent mixture such as THF/water.
[000228] Step 3: Compound (xxxix) can be converted to compound (xiii) by generating the enolate of (xxxix) with a base such as LDA in a solvent such as THF at a low temperature such as -78°C under an inert atmosphere and further treatment with a reagent (R2-LG) bearing an appropriate leaving group (eg LG = triflate). EXAMPLES
[000229] The invention is also defined in the following Examples. It is to be understood that the Examples are given by way of illustration only. From the foregoing discussion and Examples, one skilled in the art can ascertain the essential features of the invention, and without departing from the spirit and scope, can make various changes and modifications to adapt the invention to various uses and conditions. As a result, the invention is not limited by the illustrative examples mentioned hereinbelow, but is preferably defined by the appended claims thus far. Abbreviations AcOH acetic acid ACN acetonitrile AlMe3 trimethylaluminium Boc tert-butyloxycarbonyl DCC 1,3-dicyclohexylcarbodiimide DCM dichloromethane DEA diethylamine DMAP dimethylaminopyridine DME dimethyl ether DMF dimethylformamide DMSO dimethyl sulfoxide EDC 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride EDCI 1 -(3-dimethylaminopropyl)-3-ethylcarbodiimide Et2AlCl diethylaluminum chloride EtOAc ethyl acetate H2SO4 sulfuric acid HCl hydrochloric acid HOBT hydroxybenzotriazole HPLC High Performance Liquid Chromatography hr hour(s) IPA isopropyl alcohol LCMS Liquid Chromatography-Mass Spectroscopy LDA di- lithium isopropylamide LiHMDS lithium bis(trimethylsilyl)amide Me methyl MeOH methanol min minute(s) MTBE methyl tert-butyl ether N2 nitrogen NaHMDS sodium bis(trimethylsilyl)amide Pd/C palladium on carbon Ph phenyl RT retention time sat saturated TBTU O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate TEA triethylamine Tf2O trifluoromethyl anhydride ylsulfonic TFA trifluoroacetic acid THF tetrahydrofuran Example 1 (2R,3S)-N-((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin -3-yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide
Preparation 1A: tert-butyl 5,5,5-trifluoropentanoate

[000230] To a stirred solution of 5,5,5-trifluoropentanoic acid (5 g, 32.0 mmol) in THF (30 mL) and hexane (30 mL) at 0 °C was added 2,2,2- tert-butyl trichloroacetimidate (11.46 mL, 64.1 mmol). The mixture was stirred for 15 min at 0°C. Boron trifluoride etherate (0.406 mL, 3.20 mmol) was added, and the reaction mixture was allowed to warm to room temperature overnight. To the clear reaction mixture was added solid NaHCO3 (5 g) and stirred for 30 min. The mixture was filtered through MgSO4 and washed with hexane (200 mL). The solution was allowed to stand for 45 min, and the resulting solid material was filtered off on the same MgSO4 filter again, washed with hexanes (100 mL) and concentrated under reduced pressure without heating. The volume was reduced to about 30 mL, filtered through a clean calcined funnel, washed with hexane (5 mL), and then concentrated under reduced pressure without heating. The resulting liquid oil was filtered through a 0.45 µm nylon membrane filter disc to provide tert-butyl 5,5,5-trifluoropentanoate (6.6 g, 31.4 mmol 98% yield) as an oil. colorless: 1H NMR (400 MHz, CDCl3) δ ppm 1.38 (s, 9H) 1.74-1.83 (m, 2H) 2.00-2.13 (m, 2H) 2.24 (t, J=7.28 Hz, 2H). Preparation 1B: (4S)-4-(Propan-2-yl)-3-(5,5,5-trifluoropentanoyl)-1,3-oxazolidin-2-one

[000231] To a stirred solution of 5,5,5-trifluoropentanoic acid (5.04 g, 32.3 mmol) in DCM (50 mL) and DMF (3 drops) was added oxalyl chloride (3.4 mL, 38.8 mmol) dropwise over 5 min, and the solution was stirred until all bubbling subsided. The reaction mixture was concentrated under reduced pressure to yield a pale yellow oil. In a separate flask charged with a solution of (4S)-4-(propan-2-yl)-1,3-oxazolidin-2-one (4.18 g, 32.4 mmol) in THF (100 mL) at -78°C n-BuLi (2.5M in hexane) (13.0 mL, 32.5 mmol) was added dropwise via syringe over 5 min. After stirring for 10 min, the above acid chloride dissolved in THF (20 mL) was added via cannula over 15 min. The reaction mixture was warmed to 0 °C, and allowed to warm to room temperature, as was the heated bath and stirred overnight. To the reaction mixture was added saturated NH4Cl, and then extracted with EtOAc (2x). The combined organics were washed with brine, dried (Na2SO4), filtered and concentrated under reduced pressure. The crude material was purified by flash chromatography (Teledyne ISCO CombiFlash Rf, 5% to 60% solvent A/B=hexanes/EtOAc, REDISEP® SiO2 120g). Concentration of appropriate fractions provided Preparation 1B (7.39 g, 86%) as a colorless oil: 1 H NMR (400 MHz, CDCl 3 ) δ ppm 4.44 (1 H, dt, J=8.31, 3, 53 Hz), 4.30 (1H, t, J=8.69 Hz), 4.23 (1H, dd, J=9.06, 3.02 Hz), 2.98-3.08 ( 2H, m), 2.32-2.44 (1H, m, J=13.91, 7.02, 7.02, 4.03 Hz), 2.13-2.25 (2H, m), 1.88-2.00 (2H, m), 0.93 (3H, d, J=7.05 Hz), 0.88 (3H, d, J=6.80 Hz) . Preparation 1C: (2S,3R) 6,6,6-trifluoro-3-((S)-4-isopropyl-2-oxo-oxazolidine-3-carbonyl)-2-(3,3,3-trifluoropropyl)hexanoate )-tert-butyl, and Preparation 1D: 6,6,6-trifluoro-3-((S)-4-isopropyl-2-oxo-oxazolidine-3-carbonyl)-2-(3,3,3-trifluoropropyl ) (2R,3R)-tert-butyl hexanoate

[000232] To a stirred, cooled (-78 °C) solution of diisopropylamine (5.3 mL, 37.2 mmol) in THF (59 mL) under a nitrogen atmosphere was added n-BuLi (2.5M in hexane) (14.7 mL, 36.8 mmol), then heated to 0 °C to produce a 0.5M solution of LDA. A separate vessel was loaded with Preparation 1B (2.45 g, 9.17 mmol), the material was azeotroped twice with benzene (the air inlet of the RotoVap was equipped with a nitrogen inlet to completely exclude moisture), then toluene (15.3 mL) was added. This solution was added to a flask containing dry lithium chloride (1.96 g, 46.2 mmol). To the resulting mixture, cooled to -78 °C, LDA solution (21.0 mL, 10.5 mmol) was added and stirred at -78 °C for 10 min, heated to 0 °C for 10 min, then cooled again at -78°C. To a separate reaction vessel containing Preparation 1A (3.41 g, 16.07 mmol), similarly azeotroped twice with benzene, was added toluene (15.3 mL), cooled to -78 °C. and LDA (37.0 mL, 18.5 mmol) was added, the resulting solution was stirred at -78° for 25 min. At this time, the ester-derived enolate was transferred via cannula into the oxazolidinone enolate solution, stirred at -78 °C for an additional 5 min, at which time the septum was removed, and the bis(2-ethylhexanoyloxy ) solid powdered copper (9.02 g, 25.8 mmol) was quickly added to the reaction vessel, and the septum replaced. The vessel was immediately removed from the cold bath and immersed in a warm water bath (40 °C) with rapid vortexing with a concomitant color change from the initial turquoise to brown. The reaction mixture was stirred for 20 min, poured into 5% aqueous NH4OH (360 mL) and extracted with EtOAc (2x). The combined organics were washed with brine, dried (Na2SO4), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (Teledyne ISCO CombiFlash Rf, 0% to 60% solvent A/B=hexanes/EtOAc, REDISEP® SiO2 120 g). Concentration of appropriate fractions provided Preparation 1C (2.87 g, 66%) as a pale yellow viscous oil. 1H NMR showed the product to be a 1.6:1 mixture of 1C:1D diastereoisomers as determined by integrating multiplets at 2.74 & 2.84 ppm: 1H NMR (400 MHz, CDCl3) δ ppm 4.43-4 .54 (2H, m), 4.23-4.35 (5H, m), 4.01 (1H, ddd, J=9.54, 6.27, 3.51 Hz), 2, 84 (1H, ddd, J=9.41, 7.28, 3.64 Hz), 2.74 (1H, ddd, J=10.29, 6.27, 4.02 Hz), 2, 37-2.48 (2H, m, J=10.38, 6.98, 6.98, 3.51, 3.51 Hz), 2.20-2.37 (3H, m), 1 .92-2.20 (8H, m), 1.64-1.91 (5H, m), 1.47 (18H, s), 0.88-0.98 (12H, m) . Preparation 1E: (2R,3S)-3-(tert-Butoxycarbonyl)-6,6,6-trifluoro-2-(3,3,3-trifluoropropyl)hexanoic acid, and Preparation 1F: (2R,3R)- 3-(tert-butoxycarbonyl)-6,6,6-trifluoro-2-(3,3,3-trifluoropropyl)hexanoic

[000233] To a stirred, cooled (0 °C) solution of Preparation 1C and 1D (4.54 g, 9.51 mmol) in THF (140 mL) and water (42 mL) was added sequentially hydrogen peroxide (30 % in water) (10.3 g, 91 mmol) and LiOH (685.3 mg, 28.6 mmol), and the mixture was stirred for 1 hr. At this time, the reaction vessel was removed from the cold bath, and then stirred for 1.5 hr. The reaction was judged complete by HPLC. To the reaction mixture, saturated NaHCO3 (45 mL) and saturated Na2SO3 (15 mL) were added, and then partially concentrated under reduced pressure. The resulting crude solution was extracted with DCM (3x). The aqueous phase was acidified to pH~1-2 with 1N HCl, extracted with DCM (3x) and EtOAc (1x). The combined organics were washed with brine, dried (Na2SO4), filtered and concentrated under reduced pressure to provide a mixture of Preparation 1E and 1F (3.00 g, 86%) as a colorless oil: 1H NMR (400 MHz, CDCl3) δ ppm 2.76-2.84 (1H, m, diastereoisomer 2), 2.64-2.76 (3H, m), 2.042.35 (8H, m), 1.88-2.00 ( 4 H, m), 1.71-1.83 (4 H, m), 1.48 (9 H, s, diastereoisomer 1), 1.46 (9 H, s, diastereoisomer 2); 1H NMR showed a 1.7:1 mixture of 1E:1F by peak integration for t-butyl groups. Preparation 1E: (2R,3S)-3-(tert-Butoxycarbonyl)-6,6,6-trifluoro-2-(3,3,3-trifluoropropyl)hexanoic acid, and Preparation 1F: (2R,3R)- 3-(tert-butoxycarbonyl)-6,6,6-trifluoro-2-(3,3,3-trifluoropropyl)hexanoic

[000234] To a stirred, cooled (-78 °C) solution of diisopropylamine (1.7 mL, 11.93 mmol) in THF (19 mL) under a nitrogen atmosphere was added n-BuLi (2.5M in hexanes) (4.8 mL, 12.00 mmol). The mixture was stirred for 5 min, then warmed to 0 °C. In a separate vessel, into a stirred, cooled (-78 °C) solution of the mixture of Preparation 1E and 1F (1.99 g, 5.43 mmol) in THF (18 mL) was added the LDA solution prepared above by cannula medium slowly for 25 min. The mixture was stirred for 15 min, then warmed to room temperature (placed in a 24 °C water bath) for 15 min, then cooled again to -78 °C for 15 min. To the reaction mixture was added Et2AlCl (1M in hexane) (11.4 mL, 11.40 mmol) via syringe, stirred for 10 min, warmed to room temperature for 15 min, then cooled again to -78 °C. for 15 min. Methanol (25 mL) was added quickly, swirled vigorously while warming to room temperature, then concentrated to ~1/4 of the original volume. The mixture was dissolved in EtOAc and washed with 1N HCl (50 mL) and ice (75 g). The aqueous phase was separated, extracted with EtOAc (2x). The combined organics were washed with a mixture of KF (2.85 g between 75 mL of water) and 1N HCl (13 mL) [resulting solution pH 3-4], then with brine, dried (Na2SO4), filtered and concentrated under reduced pressure to yield a 9:1 (1E:1F) enriched diastereoisomeric mixture (as determined by 1H NMR) of Preparation 1E and Preparation 1F (2.13 g, >99%) as a pale yellow viscous oil: 1H NMR (400 MHz, CDCl 3 ) δ ppm 2.64-2.76 (2 H, m), 2.04-2.35 (4 H, m), 1.88-2.00 (2 H, m), 1.71-1.83 (2H, m), 1.48 (9H, s). Preparation 1G: (3S)-3-Amino-1-methyl-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one, and Preparation 1H: (3R)-3-Amino -1-methyl-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one

[000235] Racemic 3-Amino-1-methyl-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one (Rittle, KE et al., Tetrahedron Letters, 28(5): 521-522 (1987)) was prepared according to literature procedure. Enantiomers were separated under chiral SFC conditions using the following method: CHIRALPAK® AS-H 5x25; Mobile phase: 30% MeOH+ 0.1% DEA in CO2; Flow rate: 280 mL/min; Pressure: 10 MPa (100 bar); Temperature: 35°C.
[000236] The S-enantiomer obtained (Preparation 1G): HPLC: RT=1.75 min (30% MeOH + 0.1% DEA in CO2 in CHIRALPAK® AS-H 4.6x250 mm, 3 mL/min, 35 °C, 10 MPa (100 bar), 230 nm, 10 μl injection); 1H NMR (400MHz, CDCI3) δ ppm 7.58-7.63 (2H, m), 7.55 (1H, ddd, J=8.50, 7.11, 1.76 Hz), 7 .40-7.47 (1H, m), 7.34-7.40 (3H, m), 7.31 (1H, dd, J=7.81, 1.51 Hz), 7, 14-7.22 (1H, m), 4.46 (1H,s), 3.44 (3H,s), 3.42 (2H,s); [δ]D = -155° (c=1.9, MeOH) (Lit. Rittle, KE et al., Tetrahedron Letters, 28(5):521-522 (1987): [δ]D = -236°) .
[000237] Similarly obtained, the R-enantiomer (Preparation 1H): HPLC: RT = 1.71 min; [δ]D = +165° (c=2.1, MeOH) (Lit [δ]D = +227°). Alternate procedure to prepare Preparation 1G:
[000238] Preparation of 1G<SA salt: (3S)-3-amino-1-methyl-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one, acid salt (1S)-(+)-10- camphorsulfonic

[000239] The 1G<SA preparation was prepared from racemic 3-amino-1-methyl-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one (9.98g , 37.6 mmol) (prepared according to the literature as shown above) according to the literature procedure (Reider, PJ et al., J. Org. Chem., 52:955-957 (1987)). The preparation of 1G<SA (16.91 g, 99%) was obtained as a colorless solid: Optical Rotation: [δ]D = -26.99° (c=1, H2O) (Lit. [δ]D = -27.8° (c=1, H2O)) Preparation 1I: (2S,3R)-β,β,6-trifluoro-3-(((3S)-1-methyl-2-oxo-5-phenyl- tert-Butyl 2,3-dihydro-1H-1,4-benzodiazepin-3-yl)carbamoyl)-2-(3,3,3-trifluoropropyl)hexanoate and Preparation 1J: (2R,3R)-β ,β,β-trifluoro-3-(((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)carbamoyl)- tert-butyl 2-(3,3,3-trifluoropropyl)hexanoate

[000240] To a stirred solution of Preparation 1G (1.45 g, 5.47 mmol) and a 9:1 mixture of Preparation 1E and 1F (1.989 g, 5.43 mmol) in DMF (19 mL) were added tetrafluoroborate of O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium (1.79 g, 5.57 mmol) and triethylamine (3.0 mL, 21.52 mmol) and stirred during the night. The reaction was judged complete by LCMS. The reaction mixture was poured into water (125 mL), and the precipitated solid was collected by filtration, washed with water and air dried to provide an 8:1 mixture of Preparation 1I and Preparation 1J (2.95 g, 89%) as a cream solid: MS (ES): m/z = 614 [M+H]+; 1H NMR (400 MHz, CDCl3) δ ppm 7.55-7.65 (3H, m), 7.44-7.52 (2H, m), 7.35-7.45 (4H, m ), 5.52 (1H, d, J=8.03 Hz), 3.48 (3H, s), 2.63 (2H, ddd, J=9.35, 3.95, 3, 76 Hz), 2.14-2.25 (4 H, m), 1.90-2.03 (3 H, m), 1.69-1.82 (1 H, m), 1.51 ( 9 H, s). Preparation 1K: (2S,3R)-6,6,6-Trifluoro-3-(((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1 acid, 4-Benzodiazepin-3-yl)carbamoyl)-2-(3,3,3-trifluoropropyl)hexanoic acid and Preparation 1L: (2R,3R)-6,6,6-Trifluoro-3-(((3S)- 1-Methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)carbamoyl)-2-(3,3,3-trifluoropropyl)hexanoic

[000241] To a stirred, cooled (0 °C) solution of the above mixture of Preparation 1I and Preparation 1J (2.95 g, 4.81 mmol) in DCM (20 mL) was added TFA (20 mL, 260 mmol) . The reaction mixture was stirred for 1 h, then allowed to warm to room temperature and stirred for 2.5 h. The reaction was judged complete by LCMS. The reaction mixture was diluted with toluene (50 mL) and concentrated under reduced pressure. The residue mixture was redissolved in toluene (50 mL) and concentrated under reduced pressure, then dried under high vacuum. The crude product was dissolved in DCM, SiO2 (15 g) was added, concentrated, then purified by flash chromatography (Teledyne ISCO CombiFlash Rf, 0% to 45% solvent A/B=DCM/EtOAc, REDISEP® SiO2 80 g). Concentration of appropriate fractions provided a mixture of Preparation 1K and Preparation 1L (2.00 g, 75%) as a cream solid: HPLC: RT=2.770 min (CHROMOLITH® SpeedROD 4.6 x 50 mm (grad. 4 min) eluting with 1090% aqueous MeOH for 4 minutes containing 0.1% TFA, 4 mL/min, monitoring at 254 nm); MS (ES): m/z = 558 [M+H]+; 1H NMR (400 MHz, CDCl 3 ) δ ppm 8.32 (1 H, d, J=8.03 Hz), 7.65-7.71 (1 H, m), 7.50-7.60 (3 H, m), 7.41-7.49 (2H, m), 7.39 (1H, dd, J=7.91, 1.63Hz), 7.23-7.35 (2H , m), 5.59 (1H, d, J=8.03 Hz), 3.51 (3H, s), 2.81 (1H, ddd, J=10.54, 6.90, 3.64 Hz), 2.67-2.76 (1H, m), 2.22-2.33 (3H, m), 1.99-2.12 (3H, m), 1. 85-1.94 (1H, m), 1.79 (1H, ddd, J=13.87, 7.84, 3.64 Hz). Example 1:
[000242] To a stirred solution of a 8:1 mixture of Preparation 1K and Preparation 1L (3.46 g, 6.21 mmol) in DMF (25 mL) under a nitrogen atmosphere was added ammonium chloride (3.32 g , 62.1 mmol), EDC (3.55 g, 18.52 mmol), HOBT (2.85 g, 18.61 mmol) and triethylamine (16 mL, 115 mmol) and stirred overnight. The reaction was judged complete by LCMS. The reaction mixture was poured into water (200 mL) with vigorous swirling, then allowed to stand. The solid was collected by filtration, washed with water, allowed to dry to give 3.6 g of colorless solid. The solid was purified by preparative SFC chromatography (Lux-Cellulose-2 (3x25cm), 8% methanol in CO2, 140 ml/min @ 220 nm and 35 °C; Sample: 3.6 g in 50 cm3 of methanol, conc. = 70 mg/ml, Stack injection: 0.5 cm 3 / 9.2 min). The product-containing fractions were concentrated, dried overnight under vacuum. Example 1 obtained (2.74 g, 79%) as a colorless solid (Crystal Form N-1): HPLC: RT=9.601 min (H2O/CH3CN with TFA, Sunfire C18 3.5 µm, 4.6x150 mm, 4.6x150 mm, gradient = 15 min, wavelength = 220 and 254 nm). MS (ES): m/z = 557 [M+H]+; 1H NMR (400MHz, DMSO-d6) δ ppm 9.54 (1H, d, J=7.28Hz), 7.71-7.80 (1H, m), 7.68 (2H, d, J=8.78 Hz), 7.50-7.62 (3H, m), 7.45 (2H, t, J=7.28 Hz), 7.29-7.40 (2 H, m), 7.15 (1H, br.s.), 5.30 (1H, d, J=7.28 Hz), 3.39 (3H, s), 2.74-2 .86 (1H, m), 2.02-2.32 (3H, m), 1.45-1.79 (4H, m); [δ]D = -107.0° (5.73 mg/ml, DMSO).
[000243] Crystal form A-2 was prepared by adding approximately 1 mg of Example 1 to approximately 0.7 ml of acetone/acetonitrile/water (2:2:1) solution. A mixture of colorless needles and thinly sliced crystals was obtained after one day of slow evaporation of the solution at room temperature. The thinly sliced crystals were separated to give Crystal Form A-2.
[000244] Crystal form EA-3 was prepared by adding approximately 1 mg of Example 1 to approximately 0.7 ml of ethyl acetate/heptane (1:1) solution. Crystals on colorless slides were obtained after three days of slow evaporation of the solution at room temperature.
[000245] The THF-2 crystal form was obtained by adding approximately 5 mg of Example 1 to approximately 0.7 ml of THF/water (4:1) solution. Colorless blade-like crystals were obtained after one day of solvent evaporation at room temperature. Alternate procedure to prepare Example 1: Preparation 1M: 3,3,3-trifluoropropyl trifluoromethanesulfonate

[000246] To a cooled (-25 °C), stirred solution of 2,6-lutidine (18.38 mL, 158 mmol) in CH2Cl2 (120 mL) was added Tf2O (24.88 mL, 147 mmol) for 3 min, and stirred for 5 min. To the reaction mixture was added 3,3,3-trifluoropropan-1-ol (12 g, 105 mmol) over an interval of 3 min. After 2 h, the reaction mixture was warmed to room temperature and stirred for 1 h. The reaction mixture was concentrated to half volume, then purified by loading directly onto a silica gel column (330 g ISCO) and eluted with CH2Cl2. Preparation 1M obtained (13.74 g, 53%) as a colorless oil. 1H NMR (400 MHz, CDCl 3 ) δ ppm 4.71 (2H, t, J=6.15 Hz), 2.49-2.86 (2H, m). Preparation 1N: (4S)-4-Benzyl-3-(5,5,5-trifluoropentanoyl)-1,3-oxazolidin-2-one

[000247] Preparation 1N was prepared from 5,5,5-trifluoropentanoic acid (3.35 g, 21.46 mmol) and (4S)-4-benzyl-1,3-oxazolidin-2-one (3 .80 g, 21.46 mmol) by the general methods shown for Preparation 1B. Preparation 1N (5.67 g, 84%) was obtained as a colorless viscous oil: 1 H NMR (400 MHz, CDCl 3 ) δ ppm 7.32-7.39 (2 H, m), 7.30 (1 H , d, J=7.05 Hz), 7.18-7.25 (2H, m), 4.64-4.74 (1H, m), 4.17-4.27 (2H, m). m), 3.31 (1H, dd, J=13.35, 3.27Hz), 3.00-3.11 (2H, m), 2.79 (1H, dd, J=13 .35, 9.57 Hz), 2.16-2.28 (2H, m), 1.93-2.04 (2H, m). Preparation 1O: tert-(3R)-3-(((4S)-4-benzyl-2-oxo-1,3-oxazolidin-3-yl)carbonyl)-6,6,6-trifluorohexanoate butyl

[000248] To a stirred, cooled (-78 °C) solution of Preparation 1N (3.03 g, 9.61 mmol) in THF (20 mL) was added NaHMDS (1.0M in THF) (10.6 mL , 10.60 mmol) under nitrogen atmosphere. After 2 hours, tert-butyl 2-bromoacetate (5.62 g, 28.8 mmol) was added to liquid via syringe at -78°C, and stirring was continued at the same temperature. After 6 hours, the reaction mixture was warmed to room temperature. The reaction mixture was partitioned between saturated NH4Cl and EtOAc. The organic phase was separated, and the aqueous was extracted with EtOAc (3x). The combined organics were washed with brine, dried (Na2SO4), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (Teledyne ISCO CombiFlash Rf, 5% to 100% solvent A/B=hexanes/EtOAc, REDISEP® SiO2 120 g). Concentration of appropriate fractions provided Preparation 1O (2.79 g, 67.6%) as a colorless viscous oil: 1H NMR (400 MHz, CDCl3) δ ppm 7.34 (2H, d, J=7.30 Hz), 7.24-7.32 (3H, m), 4.62-4.75 (1H, m, J=10.17, 6.89, 3.43, 3.43 Hz), 4.15-4.25 (3H, m), 3.35 (1H, dd, J=13.60, 3.27Hz), 2.84 (1H, dd, J=16.62, 9.57 Hz), 2.75 (1H, dd, J=13.35, 10.07 Hz), 2.47 (1H, dd, J=16.62, 4.78 Hz), 2, 11-2.23 (2H, m), 1.90-2.02 (1H, m), 1,721.84 (1H, m), 1.44 (9H, s). Preparation 1P: (2R)-2-(2-tert-Butoxy-2-oxoethyl)-5,5,5-trifluoropentanoic acid

[000249] Preparation 1P was prepared from Preparation 1O (2.79 g, 6.50 mmol) by the general methods shown for Preparation 1E. Preparation 1P (1.45 g, 83%) was obtained as a colorless oil: 1 H NMR (400 MHz, CDCl 3 ) δ ppm 2.83-2.95 (1 H, m), 2.62-2.74 (1H, m), 2.45 (1H, dd, J=16.62, 5.79 Hz), 2.15-2.27 (2H, m), 1.88-2.00 ( 1H, m), 1.75-1.88 (1H, m), 1.45 (9H, s). Preparation 1E: (2R,3S)-3-(tert-Butoxycarbonyl)-6,6,6-trifluoro-2-(3,3,3-trifluoropropyl)hexanoic acid, and Preparation 1F: (2R,3R)- 3-(tert-butoxycarbonyl)-6,6,6-trifluoro-2-(3,3,3-trifluoropropyl)hexanoic

[000250] To a stirred, cooled (-78 °C) solution of Preparation 1P (5.44 g, 20.13 mmol) in THF (60 mL) was slowly added LDA (24.60 mL, 44.3 mmol) for 7 min. After stirring for 2 h, Preparation 1M (6.44 g, 26.2 mmol) was added to the reaction mixture over 3 min. After 45 min, the reaction mixture was heated in a -25 °C bath (ice/MeOH/dry ice) for 1 h, then warmed to 0 °C. After 45 min, the 1M preparation (1 g) was added, and the reaction mixture was stirred for 20 min. The reaction was quenched with water and 1N NaOH and extracted with CH2Cl2. The organic layer was extracted again with 1N NaOH (2x), and the aqueous layers were combined. The aqueous layer was cooled in an ice/water bath, then acidified with concentrated HCl to pH 2. Then, the aqueous layer was extracted with EtOAc. The combined organics were washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was dried under high vacuum to provide a 1:5 (1E:1F) mixture (as determined by 1H NMR) of Preparation 1E and Preparation 1F (5.925 g, 80%) as a pale yellow solid. 1H NMR (500MHz, CDCl3) δ ppm 2.81 (1H, ddd, J=10.17, 6.32, 3.85Hz), 2.63-2.76 (1H, m), 2 .02-2.33 (4H, m), 1.86-1.99 (2H, m), 1.68-1.85 (2H, m), 1.47 (9H, s) . Preparation 1E: (2R,3S)-3-(tert-Butoxycarbonyl)-6,6,6-trifluoro-2-(3,3,3-trifluoropropyl)hexanoic acid and Preparation 1F: (2R,3R)-3 acid -(tert-butoxycarbonyl)-6,6,6-trifluoro-2-(3,3,3-trifluoropropyl)hexanoic

[000251] A mixture of Preparation 1E and Preparation 1F (64 mg, 1.758 mmol) was taken up in THF (6 mL) to yield a colorless solution, which was cooled to -78 °C. Then, LDA (2.149 mL, 3.87 mmol) (1.8M in heptane/THF/ethyl benzene) was slowly added to the reaction mixture over 10 min. After stirring for 15 min, the reaction mixture was placed in a water bath at room temperature. After 15 min, the reaction mixture was placed back into a -78 °C bath, then diethylaluminum chloride (3.87 mL, 3.87 mmol) (1M in hexane) was slowly added over 5 min. The reaction mixture was stirred at -78°C. After 15 min, the reaction mixture was placed in a bath with water at room temperature for 10 min, and then cooled again in a bath to -78 °C. After 15 min, the reaction was quenched with MeOH (8 mL, 198 mmol), removed from the -78 °C bath and concentrated. To the reaction mixture, ice and HCl (16 mL, 16.00 mmol) were added, followed by extraction with EtOAc (2x). The organic layer was washed with potassium fluoride (920 mg, 15.84 mmol) (in 25 mL of H 2 O) and HCl (4.5 mL, 4.50 mmol). The organics were dried over anhydrous magnesium sulfate and concentrated under reduced pressure to provide a 9:1 (1E:1F) enriched mixture of Preparation 1E and Preparation 1F (540 mg, 1.583 mmol, 90% yield) as a yellow solid. /light orange. 1H NMR (400 MHz, CDCl 3 ) δ ppm 2.64-2.76 (2H, m), 2.04-2.35 (4H, m), 1.88-2.00 (2H, m ), 1.71-1.83 (2H, m), 1.48 (9H, s). It was converted to Example 1 by the sequence of reactions as outlined above. Alternate procedure to prepare Preparation 1E: Preparation 1Q: (2R,3S)-1-benzyl 4-tert-butyl 2,3-bis(3,3,3-trifluoropropyl)succinate

[000252] A clean and dry 5 L round bottom flask equipped with a mechanical stirrer, thermometer socket and room temperature nitrogen bubbler was charged with N,N-dimethylformamide (2.07 L), a mixture 1.2:1 of Preparation 1E and Preparation 1F (207 g, 0.5651 mol), potassium carbonate (117.1 g, 0.8476 mol) followed by benzyl bromide (116 g, 0.6781 mol) for 15-20 min. The reaction mixture was stirred for 2-3 h. After completion of the reaction, the reaction mixture was concentrated to dryness at 50-55 °C under vacuum. Ethyl acetate (3.1 L, 30 Vol.) was charged to the concentrated reaction mass, and then washed with water (2.07 L), brine (0.6 L), then dried over anhydrous sodium sulfate. (207 g), filtered and concentrated to dryness at 40-45°C under vacuum. The residue was dissolved in dichloromethane (1.035 L, 5 vol.), then absorbed onto silica gel (60-120) (607 g, 3.0 w/w), then purified with column chromatography using ether of petroleum and ethyl acetate as solvents. After grouping several batches, Preparation 1Q (235 g) was obtained. HPLC Purity: 99.77%, Preparation 1E: (2R,3S)-3-(tert-butoxycarbonyl)-6,6,6-trifluoro-2-(3,3,3-trifluoropropyl)hexanoic acid

[000253] A clean, dry 2 L autoclave was charged with methanol (540 mL), and purged with nitrogen for 5-10 minutes. To the autoclave, 10% palladium on carbon (12 g, 20%) was added, purged with nitrogen once more for 5-10 min, then charged with Preparation 1Q (60 g, 0.1315 mol), at The autoclave was stimulated with methanol (60 mL) and stirred for 4-6 hr at 20-25 °C under 5 kg of hydrogen pressure. After completion of the reaction, the reaction mass was filtered through CELITE®, washed with methanol (180 mL), dried over anhydrous sodium sulfate (60 g), filtered and concentrated to dryness at 45-50 °C under vacuum. Preparation 1E obtained (45.8 g, 95%) as a colorless solid: HPLC purity: 98.9%. Alternate procedure to prepare Preparation 1E: Preparation 1E: (2R,3S)-3-(tert-butoxycarbonyl)-6,6,6-trifluoro-2-(3,3,3-trifluoropropyl)hexanoic acid

[000254] Preparation 1E was prepared in an identical procedure as above, from a mixture of Preparations 1E and 1F (200 g, 0.5460 mol) using LDA (1.8 M solution in THF, ethyl benzene and heptane ) (698 mL, 2.3 equiv.) and diethylaluminum chloride (1.0 M solution in hexane) (1256 mL, 2.3 equiv) in THF (2.0 L). After preparation as explained above, the resulting residue was treated as follows: The raw material was added to a 2L four-necked round bottom flask, followed by the addition of MTBE (1.0L) loaded under 30°C. The resulting mixture was stirred for 5-10 minutes to obtain a clear solution. Hexanes (600 mL) were charged to the reaction mixture at a temperature below 30°C. The reaction mixture was stirred for 10 min. Then tert-butylamine (43.8 g, 1.1 eq) was slowly charged over a 15 minute period below 30°C. This addition was observed to be exothermic. The reaction mixture was stirred for 2 hrs below 30°C and filtered. The solid material was washed with 5:3 MTBE:hexane (200 mL), the filtrate was concentrated and transferred to an amber colored bottle. The filtered solid was dissolved in dichloromethane (2.0L), washed with 1N HCl (2.0), the organic layer was washed with brine (1.0L x 2), then concentrated under reduced pressure below 45° Ç. This material was found to be 91.12% pure. The material was repurified by the above t-butylamine crystallization/purification procedure. Preparation 1E obtained (78 g, 39%): HPLC purity: 99.54%. Alternate procedure to prepare Example 1: Preparation 1I: (2S,3R)-6,6,6-trifluoro-3-(((3S)-1-methyl-2-oxo-5-phenyl-2,3-di tert-butyl -hydro-1H-1,4-benzodiazepin-3-yl)carbamoyl)-2-(3,3,3-trifluoropropyl)hexanoate

[000255] A clean and dry 2 L round bottom flask equipped with a mechanical stirrer, thermometer socket and nitrogen bubbler was charged with N,N-dimethylformamide (457 mL), Preparation 1E (45.7g, 0.1248 mol) and Preparation 1G<SA (62.08 g, 0.1248 mol) under nitrogen atmosphere at 20-25°C. The reaction mixture was stirred for 15-20 minutes to prepare the clear solution at 20-25°C. To the reaction mixture was added TBTU (48.16 g, 0.1498 mol) at 20-25°C, followed by triethylamine (50.51 g, 0.4992 mol) over 15-20 minutes at 20-25°C. The reaction mixture was stirred for 60-120 minutes at 20-25 °C under a nitrogen atmosphere. Upon completion of the reaction, the reaction was quenched in water (1.37L, 30 Vol.) at 20-25°C with stirring. The reaction mixture was stirred for 30 minutes at 20-25°C. The reaction mixture was filtered and washed with water (228 mL). The resulting solid material was dissolved in ethyl acetate (457ml), washed with water (2x137ml), brine (137ml), and then dried over anhydrous sodium sulfate (45.7g). Activated carbon (9.14 g, 20%) was charged to the reaction mixture and stirred for 30 minutes. The mixture was filtered through a CELITE® bed and 1 micron filter cloth, bed of charcoal washed with ethyl acetate (137 mL), concentrated to 1.0 Vol., and then petroleum ether (457 mL , 10 Vol.) was charged and stirred for 30 minutes at 20-25°C. The solid was collected by filtration, washed with petroleum ether (137 mL), and then dried under vacuum at 40-45 °C for 8 hr until loss on drying was less than 3.0%. Preparation 1I obtained (65.2 g, 85%): HPLC purity: 98.26%. Preparation 1K: (2S,3R)-6,6,6-Trifluoro-3-(((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1 acid, 4-benzodiazepin-3-yl)carbamoyl)-2-(3,3,3-trifluoropropyl)hexanoic

[000256] A clean and dry 3 L round bottom flask equipped with a mechanical stirrer, thermometer socket and nitrogen bubbler was charged with dichloromethane (980 mL) under a nitrogen atmosphere, followed by Preparation 1I ( 140 g, 0.2282 mol) at 20-25°C. The reaction mixture was cooled to 0-5 °C and trifluoroacetic acid (980 mL) was slowly charged over 30-40 minutes. The resulting mixture was stirred for 2 hr at 0-5 °C under a nitrogen atmosphere. The reaction temperature was raised to 20 to 25 °C, and the reaction mixture was stirred for 1-2 hr at 20 to 25 °C. Upon completion of the reaction, the reaction mixture was concentrated to dryness at 50 to 55 °C under vacuum. Toluene (3x700 mL) was charged to the concentrated reaction mass, then distilled at 50 to 55 °C under vacuum. After complete concentration of toluene, ethyl acetate (280 mL) was charged to the reaction mass at 20 to 25 °C, stirred for 60 minutes, then the solid was collected by filtration, washed with ethyl acetate (140 mL) , dried under vacuum at 50 to 55°C for 12 hr until loss on drying was less than 2.0%. Preparation 1K obtained (106 g, 84%): HPLC purity: 98.43%. Example 1:
[000257] A reaction vessel was charged with Preparation 1K (30 g, 53.81 mmol), HOBt (8.7g, 64.38 mmol) and THF (150 mL) at room temperature. To the homogeneous solution was added EDCI (12.4g, 64.68 mmol), stirred for 15 min, then cooled to 8 °C. To the reaction mixture was added ammonia (2M in IPA) (81 mL, 162 mmol) over 5 min to maintain a temperature below 10 °C. The resulting heavy suspension was stirred for 10 min, warmed to room temperature for 30 min, then stirred for 4 h. Upon completion of the reaction, water (230 mL) was added over 15 min slowly to maintain a temperature below 20 °C, and then stirred for 2 h. The solid was collected by filtration, washed with water (3X60 mL), then dried under vacuum 48 hr at 55 °C. The above crude product was loaded into a 1 L 3-necked round flask. IPA (200 mL) was added, then heated to 80 °C, resulting in a homogeneous solution. Water (170 mL) was added slowly (15 min) to maintain an internal temperature >75 °C. The resulting suspension was stirred and cooled to room temperature for 2 h. The solid was collected by filtration, washed with water (2 X 50 mL), then dried under vacuum (55 °C for 24 h, and 30 °C for 48 h). Example 1 obtained (23.4 g, 78% yield): HPLC purity: 99.43%. Example 2 (2R,3S)-N-((3S)-2-Oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3-bis( 3,3,3-trifluoropropyl)succinamide
Preparation 2A: (3S)-3-Amino-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one and Preparation 2B: (3R)-3-Amino-5- phenyl-1,3-dihydro-2H-1,4-benzodiazepine-2-one

[000258] Racemic 3-Amino-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one (J. Med. Chem., 49:2311-2319 (2006), compound # 5) was prepared according to the literature procedure. Enantiomers were separated on Berger SFC MGIII Column: Lux 25X3 cm, 5 cm; Mobile phase: 30% MeOH+ 0.1% DEA in CO2; Flow rate: 150 mL/min; Temperature: 40°C; Detector wavelength: 250 nM. The S enantiomer was obtained. Preparation 2A as a white solid: 1H NMR (400 MHz, DMSO-d6) δ ppm 10.67 (1H, br.s.), 7.58 (1H, td, J=7, 65, 1.76 Hz), 7.37-7.53 (5H, m), 7.23-7.30 (2H, m), 7.147.22 (1H, m), 4.23 ( 1H, s), 2.60 (2H, br.s.); HPLC: RT = 3.0625 min (30% MeOH + 0.1% DEA in CO2 on OD-H Column, 3 mL/min, 35 °C, 9.6 MPa (96 bar), 230 nm, inj . of 10 µl); [δ]D = -208.3° (5.05 mg/mL, MeOH). In the same way the R enantiomer is obtained. Preparation 2B as an off-white solid: HPLC: RT = 3.970 min; [δ]D = 182.1° (2.01 mg/mL, MeOH). Preparation 2C: (2S,3R)-6,6,6-trifluoro-3-(((3S)-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3 tert-butyl -yl)carbamoyl)-2-(3,3,3-trifluoropropyl)hexanoate and Preparation 2D: (2R,3R)-6,6,6-trifluoro-3-(((3S)- tert-Butyl 2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)carbamoyl)-2-(3,3,3-trifluoropropyl)hexanoate

[000259] Preparation 2C was prepared from Preparation 2A (564 mg, 2.244 mmol) and a mixture of Preparation 1E and Preparation 1F (822 mg, 2.244 mmol) according to the general procedure shown for Preparation 1I. Preparation 2C obtained and Preparation 2D (1.31 g, 97%): HPLC: RT = 3.443 min (CHROMOLITH® ODS 4.6 x 50 mm (4 min gradient) eluting with 10-90% aqueous MeOH for 4 minutes containing 0.% TFA, 4 mL/min, monitoring at 220 nm); MS (ES): m/z = 600.3 [M+H]+. Preparation 2E: (2S,3R)-6,6,6-Trifluoro-3-(((3S)-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin- 3-yl)carbamoyl)-2-(3,3,3-trifluoropropyl)hexanoic acid and Preparation 2F: (2R,3R)-6,6,6-Trifluoro-3-(((3S)-2-oxo- 5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)carbamoyl)-2-(3,3,3-trifluoropropyl)hexanoic

[000260] A mixture of Preparation 2E and Preparation 2F was prepared from a mixture of Preparation 2C and Preparation 2D (1.31 g, 2.185 mmol) by the general methods shown for Preparation 1K. A mixture of Preparation 2E and Preparation 2F (1.18 g, 99%) is obtained: HPLC: RT = 2.885 min (CHROMOLITH® ODS 4.6 x 50 mm (4 min gradient) eluting with 10-90% MeOH aqueous solution for 4 minutes containing 0.% TFA, 4 mL/min, monitoring at 220 nm). MS (ES): m/z = 544.2 [M+H]+. Example 2:
[000261] Example 2 was prepared from a mixture of Preparation 2E and Preparation 2F (354 mg, 0.651 mmol) by the general methods shown for Example 1. After separation of the diastereoisomers, Example 2 (188 mg, 52%) was obtained as a white solid: HPLC: RT = 9.063 min (H2O/CH3CN with TFA, Sunfire C18 3.5 µm, 4.6x150 mm, 4.6x150 mm, gradient = 15 min, wavelength = 220 and 254 nm); MS (ES): m/z = 543 [M+H]+; 1H NMR (400 MHz, DMSO-d6) δ ppm 10.87 (1H, br.s.), 9.50-9.55 (1H, m), 7.62-7.69 (2H, m), 7.40-7.57 (5H, m), 7.29-7.36 (2H, m), 7.22-7.28 (1H, m), 7.16 (1 H, br.s.), 5.25 (1H, d), 3.30-3.32 (1H, m), 2.75-2.86 (1H, m), 2.442.48 ( 1H, m), 2.06-2.34 (3H, m), 1.51-1.77 (4H, m); [δ]D = -114.4° (8.04 mg/mL, DMSO).
[000262] Crystal form M2-1 was prepared by adding approximately 1 mg of Example 2 to approximately 0.7 ml of MeOH/fluorobenzene (3:1) solution. Colorless plate-like crystals were obtained after 2 days of solvent evaporation at room temperature. Example 3 (2R,3S)-N-((3S)-1-Methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2- (2,2,2-trifluoroethyl)-3-(3,3,3-trifluoropropyl)succinamide
Preparation 3A: (4S)-4-(Propan-2-yl)-3-(4,4,4-trifluorobutanoyl)-1,3-oxazolidin-2-one

[000263] Preparation 3A was prepared from (4S)-4-(propan-2-yl)-1,3-oxazolidin-2-one (4.66 g, 36.1 mmol) and acid 4,4 ,4-trifluorobutanoic acid (5.02 g, 35.3 mmol) by the general methods shown for Preparation 1B. Preparation 3A was obtained as a colorless oil (3.64 g, 40%). 1H NMR (400 MHz, CDCl 3 ) δ ppm 4.44 (1 H, ddd, J=8.41, 3.51, 3.39 Hz), 4.31 (1 H, t, J=8.66 Hz ), 4.25 (1H, dd, J=9.03, 3.26 Hz), 3.13-3.32 (2H, m), 2.47-2.59 (2H, m) , 2.38 (1H, dddd, J=13.96, 7.01, 3.89Hz), 0.93 (3H, d, J=7.28Hz), 0.88 (3H, d, J=6.78 Hz). Preparation 3B: (3R)-5,5,5-trifluoro-3-(((4S)-2-oxo-4-(propan-2-yl)-1,3-oxazolidin-3-yl)carbonyl)- tert-butyl 2-(3,3,3-trifluoropropyl)pentanoate

[000264] Preparation 3B was prepared from Preparation 3A (1.04 g, 4.12 mmol) and tert-butyl 5,5,5-trifluoropentanoate (Preparation 1A) (1.55 g, 7.28 mmol ) by the general methods shown for Preparation 1C. Preparation 3B (528.3 mg, 28%) was obtained as a pale yellow viscous oil: 1 H NMR (400 MHz, CDCl 3 ) δ ppm 4.57 (1 H, ddd, J=10.54, 5.02, 1.76 Hz), 4.41-4.50 (2H, m), 4.20-4.32 (4H, m), 2.77-2.88 (3H, m), 2. 70 (1H, dt, J=9.79, 4.89 Hz), 2.38 (1H, dddd, J=10.38, 6.87, 3.64, 3.45 Hz), 2, 23-2.34 (4H, m), 2.06-2.18 (2H, m), 1.93-2.05 (2H, m), 1.69-1.81 (4H , m), 1.46 (9 H, s), 1.43 (9 H, s), 0.85-0.97 (12 H, m). Preparation 3C: (2R)-3-(tert-Butoxycarbonyl)-6,6,6-trifluoro-2-(2,2,2-trifluoroethyl)hexanoic acid

[000265] Preparation 3C was prepared from Preparation 3B (528.3 mg, 1.140 mmol) by the general methods shown for Preparation 1D. Preparation 3C (306.7 mg, 76%) was obtained as a colorless waxy solid (306.7 mg, 76%). 1H NMR (400 MHz, CDCl 3 ) δ ppm 3.08 (1H, ddd, J=8.72, 4.20, 3.89 Hz), 3.00 (1H, ddd, J=9.66, 7.03, 2.89 Hz), 2.70-2.82 (4 H, m), 2.36 (1 H, ddd, J=15.25, 10.73, 3.64 Hz), 2 .18-2.30 (2H, m), 2.12 (2H, dd, J=10.16, 5.65Hz), 1.90-2.02 (2H, m), 1. 70-1.81 (3H, m), 1.45-1.51 (18H, m). Preparation 3D: (2R,3S)-3-(tert-Butoxycarbonyl)-6,6,6-trifluoro-2-(2,2,2-trifluoroethyl)hexanoic acid

[000266] Preparation 3D was prepared from Preparation 3C (306.7 mg, 0.871 mmol) by the general methods shown to enrich the mixture of Preparation 1E and Preparation 1F. Preparation 3D (297.0 mg, 97%) was obtained as a yellow waxy solid (297.0 mg, 97%). 1H NMR (400 MHz, CDCl 3 ) δ ppm 2.99 (1H, ddd, J=9.47, 6.96, 2.89 Hz), 2.69-2.82 (2H, m), 2 .18-2.31 (2H, m), 2.06-2.18 (1H, m), 1.91-2.03 (1H, m), 1.68-1.80 (1 H, m), 1.46-1.51 (9 H, m). Preparation 3E: (2S,3R)-5,5,5-trifluoro-3-(((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4 tert-butyl -benzodiazepin-3-yl)carbamoyl)-2-(3,3,3-trifluoropropyl)pentanoate

[000267] Preparation 3E was prepared from Preparation 3D (297.0 mg, 0.843 mmol) and Preparation 1G (212.0 mg, 0.799 mmol) by the general methods shown for Preparation 1I. Preparation 3E (471.7 mg, 98%) was obtained as a brown solid (471.7 mg, 98%). MS (ES): m/z = 600 [M+H]+; 1H NMR (400 MHz, CDCl 3 ) δ ppm 7.75 (1 H, d, J=7.78 Hz), 7.54-7.64 (3 H, m), 7.43-7.51 (1 H, m), 7.34-7.43 (4H, m), 7.22-7.31 (1H, m), 5.50 (1H, d, J=7.53 Hz), 3.48 (3H, s), 2.87-2.96 (1H, m), 2.73-2.83 (1H, m), 2.60 (1H, td, J=10 .10, 3.89 Hz), 2.13-2.25 (3H, m), 1.86-2.05 (2H, m), 1.52 (9H, s). Preparation 3F: (2S,3R)-5,5,5-Trifluoro-3-(((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1 acid, 4-benzodiazepin-3-yl)carbamoyl)-2-(3,3,3-trifluoropropyl)pentanoic

[000268] Preparation 3F was prepared from Preparation 3E (466.1 mg, 0.777 mmol) by the general methods shown for Preparation 1H. Preparation 3F (451 mg, >99%) was obtained as a brown solid. MS (ES): m/z = 544 [M+H]+; 1H NMR (400 MHz, CDCl 3 ) δ ppm 8.29 (1H, d, J=7.53 Hz), 7.64 (1H, td, J=7.84, 1.63 Hz), 7.537, 60 (2H, m), 7.49 (1H, t, J=7.40 Hz), 7.33-7.46 (4H, m), 7.22-7.33 (2H, m), 5.53 (1H, d, J=7.53Hz), 3.49 (3H,s), 3.04-3.21 (2H, m), 2.692.81 (2H , m), 2.23-2.33 (2H, m), 2.07-2.19 (2H, m). Example 3:
[000269] Example 3 was prepared from Preparation 3F (446 mg, 0.821 mmol) by the general methods shown for Example 1. After separation of the diastereoisomers, Example 3 was obtained: HPLC: RT = 3.17 min ( H2O/CH3CN with TFA, Sunfire C18 3.5 µm, 4.6x150 mm, 4.6x150 mm, gradient = 15 min, wavelength = 220 and 254 nm). MS (ES): m/z = 543.3 [M+H]+; 1H NMR (400MHz, DMSO-d6) δ ppm 9.72 (1H, d, J=8.53Hz), 7.71-7.77 (1H, m), 7.667.71 (1H, m), 7.64 (1H, d, J=1.25 Hz), 7.48-7.57 (3H, m), 7.39-7.47 (2H, m), 7. 30-7.39 (2 H, m), 7.23 (1 H, s), 5.36 (1 H, d, J=8.53 Hz), 3.39 (3 H, s), 3 .12-3.23 (1H, m), 2.53-2.61 (1H, m), 2.43 (1H, td, J=10.10, 3.89 Hz), 2, 16-2.28 (1H, m), 2.02-2.16 (1H, m), 1.82-1.96 (1H, m), 1.68-1.82 (1H , m). Example 4 (2R,3S)-N-((3S)-1-Methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 3-(2,2,2-trifluoroethyl)-2-(3,3,3-trifluoropropyl)succinamide
Preparation 4A: tert-butyl 4,4,4-trifluorobutanoate

[000270] Preparation 4A was prepared from 4,4,4-trifluorobutanoic acid (4.99 g, 35.1 mmol) using the general procedure shown for Preparation 1A. Preparation 4A (5.58 g, 80%) was obtained as a colorless oil. 1H NMR (400 MHz, CDCl 3 ) δ ppm 2.472.52 (2 H, m), 2.37-2.45 (2 H, m), 1.46 (9 H, s). Preparation 4B: (3R)-6,6,6-trifluoro-3-(((4S)-2-oxo-4-(propan-2-yl)-1,3-oxazolidin-3-yl)carbonyl)- tert-butyl 2-(2,2,2-trifluoroethyl)hexanoate

[000271] Preparation 4B was prepared from Preparation 4A (1058.3 mg, 5.34 mmol) and Preparation 1B (809.2 mg, 3.03 mmol) according to the general procedure shown for Preparation 1C. Preparation 4B (690.1 mg, 49%) was obtained as a pale yellow viscous oil. 1H NMR (400 MHz, CDCl 3 ) δ ppm 4.45-4.52 (1H, m), 4.234.40 (1H, m), 4.05-4.12 (1H, m), 3, 70 (1H, t, J=6.7 Hz), 3.05 (1H, ddd, J=9.9, 5.0, 2.3 Hz), 2.99 (1H, ddd, J =11.2, 5.8, 1.8 Hz), 2.58-2.91 (2H, m), 2.38-2.49 (1H, m), 2.27-2.36 (1H, m), 2.07-2.26 (1H, m), 1.96-2.04 (1H, m), 1.85-1.94 (1H, m), 1.72 -1.82 (1H, m), 1.46 (3H, s), 0.88-0.98 (3H, m); HPLC: RT = 3.36 min (MeOH/H 2 O/0.1% TFA, CHROMOLITH® SpeedROD 4.6 x 50 mm, 4 min gradient, wavelength = 220 nm). Preparation 4C: (2R)-3-(tert-Butoxycarbonyl)-5,5,5-trifluoro-2-(3,3,3-trifluoropropyl)pentanoic acid

[000272] Preparation 4C was prepared from Preparation 4B (690.1 mg, 1.489 mmol) according to the general procedure shown for Preparation 1E. Preparation 4C (335.9 mg, 64%) was obtained as a colorless oil. 1H NMR (400 MHz, CDCl3) δ ppm 2.91-3.03 (1H, m), 2.63-2.88 (2H, m), 2.50 (1H, t, J=7 .3 Hz), 2.07-2.43 (4 H, m), 1.89-2.05 (2 H, m), 1.73-1.88 (1 H, m), 1.47 (5H, s), 1.47 (4H, s). Preparation 4D: (2R,3S)-3-(tert-Butoxycarbonyl)-5,5,5-trifluoro-2-(3,3,3-trifluoropropyl)pentanoic acid

[000273] Preparation 4D was prepared from Preparation 4C (335.9 mg, 0.954 mmol) according to the general procedure shown for enriching the mixture of Preparation 1E and Preparation 1F. Preparation 4D (277.8 mg, 83%) was obtained as a brown oil. 1H NMR (400 MHz, CDCl3) δ ppm 2.90-3.03 (1H, m), 2.64-2.88 (2H, m), 2.50 (1H, t, J=7 .3 Hz), 2.06-2.43 (3H, m), 1.89-2.03 (1H, m), 1.731.88 (1H, m), 1.47 (9H, m) s). Preparation 4E: (2S,3R)-6,6,6-trifluoro-3-(((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4 tert-butyl -benzodiazepin-3-yl)carbamoyl)-2-(2,2,2-trifluoroethyl)hexanoate

[000274] Preparation 4E was prepared from Preparation 4D (277.8 mg, 0.789 mmol) and Preparation 1G (210.2 mg, 0.792 mmol) according to the general procedure shown for Preparation 1I. Preparation 4E was obtained as a cream solid (420.2 mg, 89%). 1H NMR (400 MHz, CDCl 3 ) δ ppm 7.55-7.65 (3 H, m), 7.48 (1 H, d, J=7.5 Hz), 7.36-7.45 (4 H, m), 5.51 (1H, d, J=7.8 Hz), 3.49 (3H, s), 2.872.92 (1H, m), 2.63 (1H, s ), 2.47-2.58 (1H, m), 2.16-2.36 (2H, m), 1.93-2.06 (1H, m), 1.80 (1H , s), 1.51 (9 H, s); LCMS: RT = 4.02 min (MeOH/H 2 O/0.1% TFA, CHROMOLITH® RP-18e 2.0 x 50 mm, 4 min gradient, wavelength = 254 nm); MS (ES): m/z = 600 [M+H+]. Preparation 4F: (2S,3R)-6,6,6-Trifluoro-3-(((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1 acid, 4-benzodiazepin-3-yl)carbamoyl)-2-(2,2,2-trifluoroethyl)hexanoic

[000275] Preparation 4F was prepared from Preparation 4E (417.2 mg, 0.696 mmol) according to the general procedure shown for Preparation 1K. Preparation 4F was obtained as a solvate of TFA as an amber solid (476.0 mg, 96%). 1H NMR (400MHz, CDCl3) δ ppm 8.25 (1H, d, J=8.0Hz), 7.73-7.82 (1H, m), 7,567.67 (2H, m) , 7.34-7.54 (3H, m), 5.67 (1H, d, J=8.3 Hz), 3.49-3.60 (2H, m), 3.05- 3.13 (1H, m), 2.81-2.97 (1H, m), 2.39-2.60 (1H, m), 2.18-2.33 (1H, m ), 1.95-2.13 (1H, m); LCMS: RT = 3.43 min (MeOH/H 2 O/0.1% TFA, CHROMOLITH® RP-18e 2.0 x 50 mm, 4 min gradient, wavelength = 254 nm); MS (ES): m/z = 544 [M+H+]. Example 4:
[000276] Example 4 was prepared from Preparation 4F (476.3 mg, 0.667 mmol) according to the general procedure shown for Example 1. After separation of diastereoisomers, Example 4 was obtained as a cream solid ( 120.3 mg, 32%). HPLC: RT = 9.446 min (H2O/CH3CN with TFA, Sunfire C18 3.5 µm, 4.6x150 mm, 4.6x150 mm, gradient = 15 min, wavelength = 220 and 254 nm); MS (ES): m/z = 543 [M+H+]; 1H NMR (400MHz, DMSO-d6) δ ppm 9.56 (1H, d, J=7.03Hz), 7.82 (1H, s), 7.70-7.78 (1H, m), 7.647.70 (1H, m), 7.50-7.63 (3H, m), 7.47 (2H, d, J=7.78Hz), 7.30-7, 42 (2H, m), 7.21 (1H, s), 5.30 (1H, d, J=7.03Hz), 3.39 (3H, s), 2,672.80 (2 H, m), 2.51-2.62 (2H, m), 2.19-2.29 (1H, m), 2.07-2.21 (1H, m), 1.60 -1.72 (2H, m). Example 5 (2R,3S)-N-((3S)-1-(2H3)Methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl) -2,3-bis(3,3,3-trifluoropropyl)succinamide

[000277] To a 5 mL threaded vial were added Example 2 (50 mg, 0.092 mmol), cesium carbonate (60.1 mg, 0.184 mmol) and iodomethane-d3 (6.88 μL, 0.111 mmol) in DMF (2 mL) to make a suspension. The mixture was stirred overnight at room temperature under nitrogen. LCMS showed the reaction was complete. The reaction mixture was dissolved in 2 ml of 1:1 DMF/AcOH and purified by Preparative HPLC on Luna ODS 5 a 21.2x100 mm, which was eluted with a 7 min gradient from 10% to 100% ACN/water 0 .1% 100% TFA. The appropriate fractions were concentrated to provide Example 5 (35 mg, 65%): HPLC: RT = 3.04 min (CHROMOLITH® S5 ODS 4.6 x 50 mm column, 10-90% aqueous methanol for 4 minutes containing 0.1% TFA, 4 mL/min, monitoring at 220 nm); MS (ES): m/z = 599 [M+H]+; 1H NMR (400MHz, DMSO-d6) δ ppm 9.55 (1H, d, J=7.5Hz), 7.71-7.80 (1H, m), 7.67 (2H, d, J=8.3 Hz), 7.51-7.60 (3H, m), 7.42-7.49 (2H, m), 7.30-7.39 (2H, m ), 7.15 (1H, s), 5.30 (1H, d, J=7.3Hz), 2.75-2.87 (1H, m), 2.40-2.48 (1H, m), 2.04-2.31 (4H, m), 1.46-1.76 (4H, m). Examples 6 to 11
[000278] The compounds listed below were prepared according to the general synthetic procedure described in Example 1, using the appropriate benzodiazepinone obtained by methods known to one skilled in the art, for example Carter, MC and others, J. Med. Chem., 49:2311-2319 (2006 ). Table 6

the Xbridge Phenyl (4.6X150 mm), 3.5 microns; flow rate of 1 mL/min; gradient 10-100% B over 30 min (A:0.05% TFA in water/CH3CN (95:5), B:0.05% TFA in water/CH3CN (5:95) @ 220 and 250 nm); 30 min cycle. b Xbridge Phenyl (4.6X150 mm), 3.5 microns; flow rate of 1 mL/min; gradient 10-100% B over 15 min (A:0.05% TFA in water/CH3CN (95:5), B:0.05% TFA in water/CH3CN (5:95) @ 220 and 250 nm); 15 min cycle. Examples 12 to 18
[000279] The compounds listed below were prepared according to the general synthetic procedure described in Example 1, using the appropriate benzodiazepinone obtained by methods known to one skilled in the art, for example Carter, MC and others, J. Med. Chem., 49:2311-2319 (2006 ). Table 7
the Xbridge Phenyl (4.6X150 mm), 3.5 microns; flow rate of 1 mL/min; 10-100% B gradient over 12 min (A:0.05% TFA in water/CH3CN (95:5), B:0.05% TFA in water/CH3CN (5:95) @ 220 and 250 nm); 25 min cycle. b Xbridge Phenyl (4.6X150 mm), 3.5 microns; flow rate of 1 mL/min; gradient 10-100% B for 12 min (A:0.05% TFA in water/CH3CN (95:5), B:0.05% TFA in water/CH3CN (5:95) @ 220 and 250 nm); 15 min cycle. c LCMS: MeOH/H 2 O/0.1% TFA, BEH C18 2.1x50 mm 1.7u, 2 min gradient, wavelength = 254 nm. d MeOH/H 2 O/0.1% TFA, Waters Sunfire C18 2.1x30 mm, 2 min gradient, wavelength = 254 nm. and CHROMOLITH® ODS 4.6 x 50 mm (4 min gradient) eluting with 10-90% aqueous MeOH for 4 minutes containing 0.% TFA, 4 mL/min, monitoring at 220 nm. Example 19 (2R,3S)-N-((3S)-2-Oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-3-(4,4 ,4-trifluorobutyl)-2-(3,3,3-trifluoropropyl)succinamide
Preparation 19A: (2R,3R)-3-(tert-Butoxycarbonyl)-7,7,7-trifluoro-2-(3,3,3-trifluoropropyl)heptanoic acid

[000280] Preparation 19A was prepared from Preparation 9A (674 mg, 2.59 mmol) and Preparation 1P (500 mg, 1.850 mmol) according to the alternate procedure shown for Preparation 1E. Preparation 19A obtained (521 mg, 28%): MS (ES): m/z = 379 [MH]-. Preparation 19B: (2R,3S)-3-(tert-Butoxycarbonyl)-7,7,7-trifluoro-2-(3,3,3-trifluoropropyl)heptanoic acid

[000281] Preparation 19B was prepared from Preparation 19A (198 mg, 0.521 mmol) according to the general procedure shown for enriching the mixture of Preparation 1E and Preparation 1F. Preparation 19B obtained (192 mg, 98%): MS (ES): m/z = 379 [MH]-; 1H NMR (500 MHz, CDCl 3 ) δ ppm 2.65-2.72 (1H, m), 2.60 (1H, ddd, J=10.33, 8.81, 3.61 Hz), 2 .19-2.30 (2H, m), 2.06-2.16 (3H, m), 1.85-1.96 (1H, m), 1.70-1.81 (2 H, m), 1.51-1.67 (3 H, m), 1.47 (7 H, s). Preparation 19C: (2S,3R)-6,6,6-trifluoro-3-(((3S)-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl )carbamoyl)-2-(4,4,4-trifluorobutyl) tert-butyl hexanoate

[000282] Preparation 19C was prepared from Preparation 19B (45.4 mg, 0.119 mmol) and Preparation 2A (30 mg, 0.119 mmol) according to the general procedure shown for Preparation 1I. Preparation 19C obtained (82 mg, 100%): HPLC: RT = 3.475 min (CHROMOLITH® 5 u C18 4.6 x 30 mm (4 min gradient) eluting with 10-90% aqueous MeOH for 4 minutes containing 0.1% TFA, monitoring at 220 nm); MS (ES): m/z = 614 [M+H]+. Preparation 19D: (2S,3R)-6,6,6-Trifluoro-3-(((3S)-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepine- 3-yl)carbamoyl)-2-(4,4,4-trifluorobutyl)hexanoic

[000283] Preparation 19D was prepared from Preparation 19C (73 mg, 0.119 mmol) according to the general procedure shown for Preparation 1K. Preparation 19D obtained (80 mg, 100%) as a TFA solvate: HPLC: RT = 2.926 min (CHROMOLITH® 5 u C18 4.6 x 30 mm (4 min gradient) eluting with 10-90% MeOH aqueous solution for 4 minutes containing 0.1% TFA, monitoring at 220 nm). Example 19:
[000284] Example 19 was prepared from Preparation 19D (80 mg, 0.119 mmol) according to the general procedure shown for Example 1. After separation of the diastereoisomers, Example 19 (35 mg, 49%) was obtained . HPLC: RT = 2.731 min (CHROMOLITH® 5 u C18 4.6 x 30 mm (4 min grad) eluting with 10-90% aqueous MeOH for 4 minutes containing 0.1% TFA, monitoring at 220 nm) ; MS (ES): m/z = 557 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 10.82 (1H, s), 9.42 (1H, d, J=7.21Hz), 7.65 (1H, ddd, J= 8.32, 7.07, 1.53 Hz), 7.60 (1H, d, J=2.22 Hz), 7.49-7.57 (3H, m), 7.42-7 .49 (2H, m), 7.29-7.35 (2H, m), 7.20-7.28 (1H, m), 7.03 (1H, s), 5.23 (1H, d, J=7.21 Hz), 2.70-2.79 (1H, m), 2.57-2.69 (1H, m), 2.39-2.47 ( 1H, m), 2.05-2.32 (3H, m), 1.50-1.67 (3H, m), 1.40-1.49 (1H, m), 1. 24-1.39 (2H, m). Example 20 (2R,3S)-N1-((3S)-8-Methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 3-(4,4,4-trifluorobutyl)-2-(3,3,3-trifluoropropyl)succinamide

[000285] Example 20 was prepared using a sequence of reactions as outlined for Example 19 using Preparation 19B instead of Preparation 1E. The obtained diastereoisomer mixture was separated by chiral HPLC to provide Example 20. HPLC RT = 0.89 min. (BEH C18 2.1X 50 mm, 1.7 u, 0 to 100 B in 1 min with 0.5 min hold time, Flow rate = 1 ml/min, detection at 254 nm, Solvent A: 100% of water / 0.1% TFA; Solvent B: 100% ACNl / 0.1% TFA). MS (ES): m/z = 587.2 [M+H]+. Example 21 (2R,3S)-N-((3S)-9-((2-Methoxyethyl)amino)-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin- 3-yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide
Preparation 21A: (3-((4-Methoxybenzyl)(2-methoxyethyl)amino)-2-nitrophenyl)(phenyl)methanone

[000286] A mixture of (3-chloro-2-nitrophenyl)(phenyl)methanone (850mg, 3.25mmol) and 2-methoxy-N-(4-methoxybenzyl)ethanamine (3171mg, 16.24mmol) was heated at 100°C for 16 hours. The reaction mixture was partitioned between water (50 mL) and DCM (50 mL), extracted with DCM (3 X 50 mL), dried over Na2SO4 and purified using silica gel chromatography (step gradient: 30 to 50% of ethyl acetate/hexanes) to isolate Preparation 21A (780 mg, 57.1% yield) as a brown oil: LC/MS (PHENOMENEX® Luna 5 micron C18 4.6 X 30 mm, 0 to 100 B in 2 min with 1 min hold time, Flow rate = 5 ml/min, detection at 254 nm, Solvent A: 10% methanol / 90% water / 0.1% TFA; Solvent B: 10% water / 90% methanol / 0.1% TFA) RT = 2.32; MS (ES) m/z = 443.10 [M+Na]+. Preparation 21B: (2-Amino-3-((4-methoxybenzyl)(2-methoxyethyl)amino)phenyl)(phenyl)methanone

[000287] Preparation 21A (700 mg, 1.665 mmol), zinc (1089 mg, 16.65 mmol) and ammonium chloride (891 mg, 16.65 mmol) in EtOH (40 mL) and water (20 mL) was heated to 90°C for 5 minutes. The reaction mixture was filtered through CELITE®, partitioned between water/DCM, extracted 3X10 mL of DCM, dried over Na2SO4 and concentrated to isolate Preparation 21B (580 mg, 89% yield): LC/MS (PHENOMENEX® Luna 5 microns C18 4.6 X 30 mm, 30 to 100 B in 4 min with 1 min hold time, Flow rate = 5 ml/min, detection at 254 nm, Solvent A: 10% methanol / 90% water / 0.1% TFA: Solvent B: 10% water / 90% methanol / 0.1% TFA): RT = 2.47 min; MS (ES): m/z = 391.16 [M+H]+. Preparation 21C: 9-((4-Methoxybenzyl)(2-methoxyethyl)amino)-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]diazepin-3- benzyl ylcarbamate

[000288] following the general procedure for Preparation 50D (38.6% yield): LC/MS (PHENOMENEX® Luna 5 micron C18 4.6 X 30 mm, 30 to 100 B in 4 min with a hold time of 1 min, Flow rate = 5 ml/min, detection at 254 nm, Solvent A: 10% methanol / 90% water / 0.1% TFA; Solvent B: 10% water / 90% methanol / 0 .1% TFA) RT = 2.42 min; MS (ES): m/z = 579.22 [M+H]+. Example 21:
[000289] Example 21 was prepared from Preparation 21C using the general sequence of reactions as outlined for Example 1. The mixture of diastereoisomers obtained was separated by chiral HPLC to provide Example 21: LC/MS (PHENOMENEX® Luna 5 micron C18 4.6 X 30 mm, 30 to 100 B in 4 min with 1 min hold time, Flow rate = 5 ml/min, detection at 254 nm, Solvent A: 10% methanol / 90% of water / 0.1% TFA; Solvent B: 10% water / 90% methanol / 0.1% TFA) RT = 2.13 min; MS (ES): m/z = 616.22 [M+H]+. Comparative Compounds 22 to 25
[000290] Comparative Compounds 22 to 25 can be prepared according to the procedures described in US Patent No. 7,053,084 for Examples 8, 12a, 38 and 45a, respectively.
Example 26
[000291] Pharmaceutical formulation comprising (2R,3S)-N-((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3- yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide.
[000292] An injectable drug was formulated comprising (2R,3S)-N-((3S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepine- 3-yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide, Example 1, as a sterile, single-use, ready-to-use (RTU) solution for intravenous (IV) administration using 50:50 (v/v) of combinations of purified polyoxyethylated castor oil (a surfactant) and dehydrated alcohol (a solvent). The drug product was a sterile pale yellow to slightly cloudy (opalescent), colorless to pale solution, stored in 5 mL Type I flint glass vials, closed with 20 mm plugs, and sealed with 20 mm aluminum seals. The concentrated formulation may be diluted prior to administration with commonly used intravenous diluents, such as Normal Saline (NS) or 5% Dextrose, to provide a physiologically acceptable diluted product. Table 11 Concentrated Pharmaceutical Composition

[000293] Purified Polyoxyethylated Castor Oil: CREMOPHOR (BASF Corp.)
[000294] The concentrated pharmaceutical formulation was found to be stable in storage at 25°C/60% relative humidity, 40°C/75% relative humidity, and 50°C for a period of three months. Likewise, a photostability study (HIL/UVA) indicated that the product did not need to be protected from light. The concentrated formulation of Example 1 had long term shelf stability including chemical and physical stability.
[000295] Prior to IV administration, the concentrated pharmaceutical formulation was diluted with Normal Saline (NS) or 5% Dextrose in Water (D5W) at concentrations between 0.01 mg/mL and 0.06 mg/mL. Time-of-use/compatibility results indicated that the product diluted in NS or D5W at concentrations ranging from 0.01 mg/mL to 0.06 mg/mL was compatible with non-PVC, non-DEHP IV infusion bags. Results showed essentially no change for 24 hours storage at 2°C to 8°C or room temperature/room light (25°C and approximately 5000 lux). BIOLOGICAL TESTS
[000296] The pharmacological properties of the compounds of this invention can be confirmed by various biological assays. The following exemplified biological assays were performed with compounds of the invention. Notch-CBF1 Transactivation Assay
[000297] Notch-CBF1 cell-based transactivation assay (C promoter binding factor I) is based on the ability of released intracellular Notch domain fragments (NICDs) to function as transcription factors along with CBF1 and others nuclear factors. Luciferase assays were used to measure antagonism of Notch-CBF1 transcriptional activity. HeLa cervical cancer cells are transiently cotransfected with pCDNA3.1/Higro plasmids containing truncated Notch 1, Notch 2, Notch 3, or Notch 4 receptors and a PGL3 luciferase reporter vector containing 4 copies of the CBF1 binding site. Cells were then tested for Notch-CBF1 activity in the absence or presence of test compounds. HeLa cells, maintained in DMEM (high glucose with HEPES), 1X glutamine/penicillin/streptomycin, and 10% Fetal Bovine Serum, were transiently transfected into a T175 Vial (4.5 x106 cells/vial) using the Monster Transfection Kit (Mirus #MIR2906) according to manufacturers' specifications. Table 12 denotes the respective amount of DNA by the transfections. Table 12

[000298] Six hours post-transfection, cells were trypsinized and seeded into a 384-well black Poly-D-lysine coated tissue culture plate at a density of 5x103 cells/well in 95 μL of assay media (DMEM (high glucose with HEPES), 1X glutamine/penicillin/streptomycin, 0.0125% BSA, 1X non-essential amino acids). Assay media (5 μL) containing the test compound in final concentrations ranging from 5 μM to 8.4x10 -5 μM (3-fold serial dilutions) were added to the cells and the cell plates were then incubated for 18 hours at 37°C and 5% CO2. Control wells contained either DMSO vehicle (total counts) or 0.5 µM of a local small molecule inhibitor (base counts). Duplicates were used for each sample. Luciferase activity was measured after a 20 minute incubation with 50 µl of STEADY-GLO® luciferase reagents according to the manufacturer's specifications (Promega, Cat. #: E2550) and analyzed by an Envision plate reader (PerkinElmer, Boston, MA).
[000299] The antagonist effect of the compounds was expressed as 100 x [1-(median sample - median base)/(median total - median base)] where the sample is the luciferase activity in the presence of the test compound, the base is equal to the luciferase activity in the presence of the small molecule inhibitor control and the total is the signal induced in DMSO wells. Data were plotted using a four-parameter logistic fit equation and the IC50 value was defined as the concentration of compound that inhibited 50% of luciferase activity.
[000300] Table 13 below lists the Notch 1 and Notch 3 IC50 values for Examples 1 to 21 of this invention and Comparative Compounds 22 to 25 measured in the Notch-CBF1 Transactivation Assay hereinabove. The results in Table 13 have been rounded to 2 digits. Compounds of the present invention as exemplified by Examples 1 to 21 showed Notch 1 values of 6.6 nM or less and Notch 3 IC50 values of 13 nM or less. Table 13

High Processing (HT) Metabolic Stability Panel
[000301] Compounds administered parenterally enter the blood stream and pass through one or more passages through the liver. Compounds that are not readily metabolized by the liver can be administered at therapeutically effective plasma levels for therapeutically effective periods of time.
[000302] Orally administered compounds are typically absorbed through the intestinal walls into the blood stream and pass a first pass through the liver. Compounds that are not readily metabolized in this first pass through the liver can be delivered to other areas of the body in therapeutically effective amounts.
[000303] Metabolic stability assay evaluated CYP-mediated metabolic stability in vitro using human, rat, mouse, dog and/or monkey microsomes after a ten minute incubation. Each compound was tested in duplicate.
[000304] The results of these assays were expressed as the fraction of the parent compound remaining in the reaction mixture after a ten minute incubation (Percent Remaining). In general, these results were used to assess only the extent of NADPH-dependent or CYP-mediated metabolism of the test compound. When the compound was significantly metabolized (< 4050% remaining), this indicated high clearance of the compound in vivo due to CYP-mediated metabolism. However, if the compound demonstrated moderate (50-80%) or low (>85%) metabolism in these in vitro assays, high clearance was still possible in vivo through other metabolism and series of elimination reactions.
[000305] The percentage of remaining results from these assays were predictive of compound clearance in vivo, assuming that CYP-mediated metabolism was a predominant elimination reaction series. In different microsomal species, the ranges of results were approximately as shown in Table 14. Table 14 Metabolic Stability - Result Interpretation Guidelines

[000306] Table 15 below lists the CYP-mediated metabolic stability for Examples 1 to 21 of this invention and Comparative Compounds 22 to 25 measured in the metabolic stability assays in humans and mice. The results in Table 15 have been rounded to 2 digits. In liver microsome assays, a value of 0% remaining indicated complete CYP-mediated metabolism of a test compound, and a value of 100% did not indicate detectable CYP-mediated metabolism of a test compound. Compounds of the present invention as exemplified by Examples 1 to 21 had metabolic stability values of 80% or greater remaining for human liver microsomes (HLM) and 72% or greater remaining for mouse liver microsomes (MsLM) . In comparison, Comparative Compounds 22 to 25 had metabolic stability values of 39% or less remaining for human liver microsomes and 15% or less remaining for mouse liver microsomes. Table 15


[000307] The compounds of the present invention (Examples 1 to 21) were compared to Comparative Compounds 22 to 25 described in U.S. Patent. At the. 7,456,172, and have been found to be especially advantageous. The compounds of the present invention had a surprising advantage of the combination of activity as inhibitors of Notch 1 and Notch 3 and superior metabolic stability for liver microsomes. As shown in Tables 13 and 15, in the reported tests, Examples 1 to 21 of this invention had Notch 1 IC50 values of 6.6 nM or less and Notch 3 IC50 values of 13 nM or less; and metabolic stability values of 80% or greater remaining for human liver microsomes (HLM) and 72% or greater remaining for mouse liver microsomes (MsLM). In comparison, in similar tests, Comparative Compounds 22 to 25 had Notch 1 IC50 values of 5.1 nM or greater and Notch 3 IC50 values of 13 nM or greater; and metabolic stability values of 39% or less remaining for human liver microsomes and 15% or less remaining for mouse liver microsomes. Methods and Materials Incubation with Liver Microsomes
[000308] Test compound was received as a solution of 3.5 mM starting material in 100 percent DMSO. The test compound was diluted to create a 50 μM acetonitrile (ACN) solution containing 1.4% DMSO, which was then used as a 100x starting material for incubation with microsomes. Each compound was tested separately in duplicate in each of three species in the Metabolic Stability Human, Rat and Mouse test set or as individual species in the Metabolic Stability Dog or Metabolic Stability Monkey sets. Compound, NADPH and liver microsome solutions were combined for three-step incubation: 1. 152 μl of liver microsome suspension, 1.1 mg/ml protein concentration in 100 mM NaPi, pH 7.4 , 5 mM MgCl2 buffer, were preheated to 37°C. 2. 1.7 µl of 50 µM compound (98.6% ACN, 1.4% DMSO) was added to the same tube and pre-incubated at 37°C for 5 minutes. 3. The reaction was initiated by the addition of 17 μl of 10 mM NADPH solution preheated in 100 mM NaPi, pH 7.4.
[000309] The reaction components were mixed well, and 75 μl of the reaction mixture was transferred immediately into 150 μl of quench/stop solution (time point zero, T0). Reactions were incubated at 37 °C for 10 minutes and then an additional 75 µl aliquot was transferred into 150 µl of quench solution. Acetonitrile containing 100 μM of DMN (a UV standard for injection quality control) was used as the quenching solution to terminate the metabolic reactions.
[000310] The quenched mixtures were centrifuged at 1500 rpm (~500 X g) in an ALLEGRA® X-12 centrifuge, SX4750 rotor (Beckman Coulter Inc., Fullerton, CA) for fifteen minutes to pellet the denatured microsomes. A 90 μl volume of supernatant extract, containing the mixture of parent compound and its metabolites, was then transferred to a separate 96-well plate for UV-LC/MS-MS analysis to determine the percentage of the parent compound that remained in the mixture. Table 16 Metabolic Stability Assay - Reaction Components
Sample Analysis - Instrumentation
[000311] HPLC: Pump - Thermo Surveyor; Sample collector - CTC/LEAP HTS; UV detector - Thermo Surveyor PDA plus; Column - Varian C18, 3 μm, 2 x 20 mm with a 0.5 μm in-line filter; Mobile Phase for structural integrity pre-analysis: (A) 98% water, 2% acetonitrile with 10 mM ammonium acetate; (B) 10% water, 90% acetonitrile with 10 mM ammonium acetate; Mobile Phase for reaction sample analysis: (A) 98% water, 2% acetonitrile with 0.1% formic acid; (B) 2% water, 98% acetonitrile with 0.1% formic acid; (C) 0.1% ammonium hydroxide in water; (D) 0.1% ammonium hydroxide in acetonitrile.
[000312] Mass Spectrometer: Thermo TSQ Quantum Ultra triple quad-polar mass spectrometer; Sample Analysis: Structural Integrity Pre-analysis.
[000313] Metabolic Stability structural integrity pre-analysis was used to assess the purity of the compounds that are analyzed. Compounds were received in 96-well plates as 57 µl of a 3.5 mM DMSO solution. The 3.5 mM DMSO compound stock solutions were diluted 18-fold with a solution containing equal volumes of acetonitrile, isopropanol, and MilliQ-H2O. The resulting solutions (200 μM) were analyzed for structural integrity by LC-UV/MS on a Thermo LCQ Deca XP Plus ion capture mass spectrometer, using a 2 x 50 mm column, Waters XBridge C18, 5 μm with a 2.1 mm Waters Sentry guard column and the LC conditions described in the table below, with an injection of 5 μl and a flow rate of 1 ml/min. Acquired data reflected purity by UV absorbance at 220 nm. Only results for those compounds with purity greater than 50% were reported. Table 17 Metabolic Stability - Structural Integrity Gradient
Sample Analysis - Incubated Samples
[000314] The MS/MS condition optimization was conducted on a Thermo TSQ Quantum triple quadrupole mass spectrometer equipped with an automatic infusion heated electrospray (H-ESI) source to obtain the SRM transitions and their energy values of matching collision. Compound solutions at a concentration of 20 μM in 1:1 methanol:water were infused at a flow rate of 90 μL/min, then combined with the mobile phase at a flow rate of 50 μL/min before be introduced at the source. All compounds were optimized first using mobile phases A and B (50% A and 50% B), and if necessary using mobile phases C and D (similarly with a 50:50 composition). The optimized parameters, including polarity, SRM transition, and collision energy, were stored in a Microsoft Access database.
[000315] The mass spectrometric conditions obtained from automatic infusion to analyze the incubation samples of the Metabolic Stability assay. The injection volume was 5 μl and the flow rate was 0.8 ml/min. The gradient used in the table below has been shown. All samples were injected with the gradient using mobile phases A and B first. If necessary (eg, for chromatographic reasons), samples were reinjected with the same gradient, but using mobile phases C and D. All LC-MS/MS analysis parameters were captured electronically in the raw data files. . Table 18 Metabolic Stability - Sample Analysis Gradient
Data analysis
[000316] Peak integration was performed with XCALIBUR® software. The percentage of the remaining calculation was performed by comparing the LC-MS/MS peak areas of the T10min samples to those of the T0minuto samples for each compound. Quality control
[000317] A set of three compounds was tested along with the test compound on each assay plate. Data were accepted and transferred only if results for these control compounds fall within the expected ranges shown below. Table 19 Metabolic Stability Assay - Control Compound Values by Microsome Species
SD = Standard Deviation Metabolic Stability Half-Life Panel
[000318] Metabolism rate and half-life determined in vitro in human or animal liver microsomes was used to determine the intrinsic clearance (CLint) and hepatic clearance (CLh,b) of a compound. These parameters have been useful in predicting human clearance in vivo which defines the level of drug exposure in vivo (Obach et al., J. Pharmacol. Exp. Ther., 283:46-58 (1997); Obach, Drug Metab. Dispos. ., 27:1350-1359 (1999)).
[000319] The Metabolic Stability Half-Life Assay Panel evaluates the time course and rate of CYP-mediated (NADPH-dependent) metabolism in vitro in human, rat, mouse, dog, and monkey microsomes. The time course encompasses a 45 minute incubation, and includes 0, 5, 10, 15, 30, and 45 minute time points, at which each of the amount of test compound remaining in the mixture was measured. Result Interpretation Guideline
[000320] The results of these assays were expressed as a half-life (T1/2, min), and the fraction of parent compound remaining in the reaction mixture at each time point (Percent Remaining) was similarly reported. In general, these results should be used to assess only the extent of CYP-mediated or NADPH-dependent metabolism of the test compound. When the compound was significantly metabolized (T1/2 < 8-14 min), it indicated high in vivo clearance due to CYP-mediated metabolism. However, if the compound demonstrated moderate (50-80%) or low (>85%) metabolism in these in vitro assays, high clearance was still possible in vivo through other metabolism and elimination reaction series.
[000321] The results of these assays were predictive of compound clearance in vivo, assuming that CYP-mediated metabolism was a predominant elimination series. In different microsomal species, the ranges of results were approximately as shown in the following table: Table 20 Metabolic Stability Half-Life - Result Interpretation Guidelines
Methods and Materials
[000322] Liver microsomes were purchased from BD-Biosciences (Woburn, MA) and NADPH from AppliChem Inc; all other reagents were obtained from Sigma. Incubation with Liver Microsomes
[000323] Test compound was received as a 3.5 mM solution of raw material in 100 percent DMSO. The test compound was diluted to create a 50 μM acetonitrile (ACN) solution containing 1.4% DMSO, which was then used as a 100-fold starting material for incubation with microsomes. Each compound was tested in human, rat, mouse, dog and monkey liver microsomes. Compound, NADPH and in vivo microsome solutions were combined for three-step incubation: 1. 450 μl of liver microsome suspension, 1.1 mg/ml protein concentration in 100 mM NaPi, pH 7.4 , 5 mM MgCl2 buffer, were preheated to 37°C. 2. 5 µl of 50 µM compound (98.6% ACN, 1.4% DMSO) was added to the same tube and pre-incubated at 37°C for 5 minutes. 3. The reaction was initiated by the addition of 50 μl of 10 mM NADPH solution preheated in 100 mM NaPi, pH 7.4.
[000324] The reaction components were mixed well, and 65 μl was immediately transferred into 130 μl of quench/stop solution (time point zero, T0). Reactions were incubated at 37 °C for 5, 10, 15, 30 and 45 minutes and at each time point a 65 µl aliquot was transferred into 130 µl of quenching solution. Acetonitrile containing Internal Standard (100 ng/ml), was used as the quenching solution to terminate the metabolic reactions.
[000325] The quenched mixtures were centrifuged at 1500 rpm (~500 X g) in an ALLEGRA® X-12 centrifuge, SX4750 rotor (Beckman Coulter Inc., Fullerton, CA) for fifteen minutes to pellet the denatured microsomes. A 90 μl volume of supernatant extract, containing the parent compound mixture and its metabolites, was then transferred to a separate 96-well plate for LC/MS-MS analysis to determine the percent compound from origin that was left in the mix. Table 21 Metabolic Stability Half-Life Assays - Reaction Components

Sample Analysis - Instrumentation
[000326] HPLC: Pump binary pumps - Shimadzu LC-20 AD series; Sample collector - CTC/LEAP HTS.
[000327] The exemplified compounds of the invention showed the surprising advantage of low clearance due to CYP-mediated metabolism in both human (HLM) and mouse (MsLM) metabolic stability assays. The compounds of the present invention, as exemplified by Examples 1-2, 5-7, 9-14, 16, 18, 21-23, and 26, had the remaining percentage values in the range of 60% to 100% for the assay of human liver microsome, and 25% to 100% for the mouse liver microsome assay. In comparison, Comparative Compounds 60-61 had the remaining percentage values of 7.0% or less equally in the mouse and human liver microsome assays. Comparative Compounds 61-62 showed equally high clearance in human and mouse metabolic stability assays, indicating that the compounds were cleared by CYP-mediated metabolism in the liver. Human Tumor Xenograft Models in Mice
[000328] All rodents were obtained from Harlan Sprague Dawley Co. (Indianapolis, Indiana), and maintained in an ammonia-free environment in a defined, pathogen-free colony. All mice were quarantined for approximately 1 week prior to their use for tumor propagation and drug efficacy testing. Mice were fed food and water ad libitum. The Bristol-Miers Squibb Pharmaceutical research Institute animal care program is fully approved by the American Association for Acdreditation of laboratory Animal Care (AAALAC). All experiments were performed according to Bristol-Miers Squibb (BMS) animal testing guidelines and methods.
[000329] Tumor xenografts were grown and maintained subcutaneously (SC) in immunocompromised balb/c nu/nu or NOD-SCID mice (Harlan Sprague Dawley). Tumors were propagated as subcutaneous transplants into the appropriate mouse strain (Table 22) using tumor fragments obtained from the donor mouse. Table 22
[000330] Histological Types and Host Mouse Strain/Genus Requirement for Propagation of Various Human Tumor Xenografts in Mice
Preclinical Chemotherapy Experiences
[000331] Required numbers of animals needed to detect a significant response were pooled at the start of the experiment and each was given a subcutaneous implant of a tumor fragment (~20 mg) with a 13 gauge trocar. Tumors were allowed to grow to a predetermined size window (tumors outside the range were excluded) and animals were evenly distributed to various treatment and control groups. There were typically 8 mice for treatment and control groups, with the exception of experiments conducted in the SAL-IGF tumor model (this is not included in Table 22), in which there were typically 5 mice per treatment and control group. Treatment of each animal was based on individual body weight. Treated animals were checked daily for treatment related toxicity/mortality. Each group of animals was weighed before starting treatment (Weight1) and then again following the last treatment dose (Weight2). The difference in body weight (Weight2-Weight1) provides a measure of treatment-related toxicity.
[000332] Tumor response was determined by measuring tumors with a caliper twice weekly, until tumors reached a predetermined "target" size of 0.5 gm or 1 gm depending on tumor type. Tumor weights (mg) were estimated by the formula: Tumor weight = (length x width2) + 2
[000333] Tumor response criteria are expressed in terms of tumor growth inhibition (% TGI). Tumor growth delay is defined as the difference in time (days) required for the treated tumors (T) to reach a predetermined target size compared to those of the control group (C). For this purpose, the tumor weight of a group is expressed as the mean tumor weight (MTW).
[000334] Tumor growth inhibition is calculated as follows:

[000335] where, Ct = Mean control tumor size at the end of treatment C0 = Mean control tumor size at the beginning of treatment Tt = Mean tumor size of the treated group at the end of treatment T0 = Mean tumor size treated group at the start of treatment
[000336] Activity is defined as achieving durable tumor growth inhibition of 50% or greater (i.e., TGI > 50%) over a period equivalent to at least 1 tumor volume doubling time and the Drug treatment should be for a period equivalent to at least 2 tumor volume doubling times.
[000337] Tumor response was also expressed in terms of tumor growth retardation (TGD value), defined as the difference in time (days) required for treated tumors (T) to reach a predetermined target size compared to those of the tumor. control group (C).
[000338] Whenever possible, antitumor activity was determined over a range of dose levels up to the maximum tolerated dose (MTD) which is defined as the dose level immediately below which excessive toxicity (i.e. more than one death) has occurred. When death occurred, the day of death was recorded. Treated mice dying before their tumors reached the target size were considered to have died of drug toxicity. No control mice died carrying tumors smaller than the target size. Treatment groups with more than one death caused by drug toxicity were considered to have had overly toxic treatments and their data were not included in the assessment of an antitumor efficacy of the compound.
[000339] Interaction of potential drug toxicity affecting treatment tolerability is an important consideration in combination chemotherapy experiments. Interpretation of combination therapy results should be based on comparing the antitumor activity of the best possible response to the agents alone versus the combination at comparably tolerated doses. Therefore, therapeutic synergism was defined as a therapeutic effect obtained with a tolerated regimen of the combined agents that exceeded the optimal effect obtained at any tolerated dose of monotherapy. Statistical evaluations of data were performed using Gehan's generalized Wilcoxon test. Statistical significance was stated at P < 0.05. Drug Administration
[000340] In in vitro studies, all agents were dissolved in 100% DMSO and serially diluted in medium/10% fetal bovine serum. For administration of Notch inhibitors to rodents, two different excipients were used: [1] 94% Labrafil / 5% ETOH / 1% TW80 or [2] ETOH/TPGS/PEG300 (10:10:80). Notch inhibitors were topically administered orally on a scale of QDx15, 10 days on - 2 days off, although other scales had also been evaluated and shown to be effective. For example, dosing regimen consisting of QDx12, 4 days on - 3 days off was shown to be equally effective as QDx15, 10 days on - 2 days off. In vivo antitumor activity
[000341] The antitumor activity of Example 1 administered via the intravenous (IV) route was evaluated in human tumor xenografts implanted in mice. As shown in Figure 6, Example 1 exhibited antitumor activity.
[000342] Table 23 below lists the antitumor activity of examples of this invention as measured in mouse Human Tumor Xenograft Models. Compounds of the present invention, as exemplified by Examples 1 and 2, showed antitumor activity with oral (PO) administration. Table 23 Scale: QDx15, 10 days yes - 2 days no; Oral Administration
QD - once a day LCK - Cell Death Log
[000343] Example 1 demonstrates broad spectrum antineoplastic activity compared to a broad array of human cancer xenografts grown in mice. Significant antitumor activity was demonstrated in 16 human cancer xenografts, including human T-cell acute lymphoblastic leukemia, breast carcinoma, pancreatic carcinoma, ovarian carcinoma, glioblastoma, non-small cell lung carcinoma, colon carcinoma, osteogenic sarcoma, and neuroblastoma ( Table 24). Table 24

aAll treatments were PO, QDx15, 10 days on - 2 days off, in dosages ranging from 5 to 10 mg/kg/adm. Combination Chemotherapy
[000344] A series of studies were conducted to evaluate the combinability of Example 1 with various anticancer agents including dasatinib, paclitaxel, tamoxifen, dexamethasone, and carboplatin Example 1 and Dasatinib
[000345] Human T-cell lymphoblastic leukemia was used to assess the combined efficacy of Example 1 and dasatinib. Dasatinib treatment alone produced an antitumor effect of 1.7 LCK (10 mg/kg/adm, QD x 49, PO). The compound of Example 1 produced only modest activity of 0.1-0.5 LCK in the dose range of 3.75-7.5 mg/kg. However, the combination of the two agents produced synergistic antitumor activity, producing an antitumor efficacy of >>2.6 LCK that was significantly superior to the individual agent dasatinib alone (P<0.05). In addition, the combination regimen produced a complete response (CR) in 100% of the mice, whereas none of the individual agents produced CR (Figures 7-8 and Table 25). Table 25
[000346] Antitumor Efficacy by Combined Chemotherapy with Example 1 and Dasatinib in ALL-SIL T-cell Lymphoblastic Leukemia
aRegime = PO, QD x3, 7 times a week bRegime = PO, QD x 49 cTarget tumor size = 1000 mg II. Example 1 and Paclitaxel
[000347] The antitumor efficacy of Example 1 in combination with pa-clitaxel was evaluated in MDA-MB-468 breast carcinoma. Example 1 as a single agent produced 0.5-1.4 LCK in the dose range of 3.75 to 7.5 mg/kg/adm. Paclitaxel administered weekly at a dose of 12 mg/kg/adm produced 0.5 LCK (Figures 9 to 10 and Table 26). The combination of Example 1 in the dose range of 3.75 to 7.5 mg/kg/adm and paclitaxel produced 3.4 to 4.1 LCK of antitumor effects which was significantly superior to the single agent compound of Example 1 alone ( P=0.0006 and 0.0002, respectively). Table 26 Antitumor Efficacy by Combined Chemotherapy with Example 1 and Paclitaxel in MDA-MB-468 Human Breast Carcinoma.
aRegime = PO, QD x3, 7 times a week bRegime = IV, Q7D x 6 cTarget tumor size = 500 mg III. Example 1 and Tamoxifen
[000348] The antitumor efficacy of Example 1 in combination with Tamoxifen was evaluated on MCF7 ER receptor positive human breast carcinoma xenograft grown in female nu/nu mice. Example 1 as a single agent produced tumor growth inhibition (TGI) of 43 to 58% in the dose range of 3.75 to 7.5 mg/kg/adm without any CR or PR. Tamoxifen, administered at the MTD dose of 20 mg/kg/adm, IP, Q2 DX12, produced a % TGI of 78%, with no CR or PR. Combinations of compound of Example 1 and Tamoxifen were clearly synergistic producing % TGI of 101 and 99, respectively, at Example 1 doses of 7.5 and 3.75 mg/kg/adm. In addition, approximately 50% of mice receiving the combinations experienced tumor retraction such as PR or CR (Figures 11 to 12 and Table 27). Table 27
[000349] Antitumor Efficacy by Combined Chemotherapy with Example 1 and Tamoxifen in MCR7 Human Breast Carcinoma
aRegime = PO, QD x3, 3 times a week bRegime = IP, Q2D x 10 cTarget tumor size = 500 mg IV. Example 1 and Dexamethasone
[000350] The antitumor efficacy of Example 1 in combination with the glucocorticoid, dexamethasone, was evaluated on HPB-ALL human T-ALL leukemia xenografts grown in NOD-SCID mice. Example 1 as an individual agent was active in this model producing 1.1 LCK at 7.5 mpk. Dexamethasone was modestly active as a single agent producing 0.7 LCK at its MTD of 7.5 mpk. The combination of Example 1 and dexamethasone produced 1.9 LCK, significantly higher than either of the individual agents alone (Figure 13 and Table 28). Table 28
[000351] Antitumor Efficacy by Combined Chemotherapy with Example 1 and Dexamethasone in HPB-ALL Human Acute Lymphoblastic Leukemia

aRegime = PO, QD x3, 3 times per week bRegime = IP, QD x 14 cTarget tumor size = 3000 mg V. Example 1 and Carboplatin
[000352] The antitumor efficacy of Example 1 in combination with carboplatin was evaluated on PA-1 human ovarian teratocarcinoma xenograft grown in female nude/nude mice. Example 1 as a single agent produced 0.2 LCK at a dose of 1 mg/kg/adm. Carboplatin administered weekly at a dose of 90 mg/kg/adm produced 2.1 LCK (Figure 14 and Table 29). The combination of Example 1 at the dose of 1 mg/kg/adm and carboplatin produced >3.1 LCK of antitumor effect which was significantly superior to the single agent compound of Example 1 alone (P=0.004). Table 29
[000353] Antitumor Efficacy by Combined Chemotherapy with Example 1 and Carboplatin in PA-1 Human Ovarian Teratocarcinoma
aRegime = PO, QD x21 (1 mg/kg) bRegime = IV, Q7D x 3 cTarget tumor size = 500 mg Single Crystal X-Ray Diffractometry
[000354] Single crystal data were collected on a Bruker-AXS APEX2 CCD system using Cu Kα radiation (À= 1.5418 A). Indexing and processing of measured intensity data were performed with the APEX2 software program suite. When indicated, the crystals were cooled in the cold stream of an Oxford cryo system during data collection. Structures were resolved by direct methods and refined based on observed reflections using SHELXTL. The derived atomic parameters (coordinates and temperature factors) were refined through wide matrix least squares. The minimized function in the refinements was ∑w(|Fo| - |Fc|)2. R is defined as ∑ |Fo| - |Fc|/∑ |Fo| whereas Rw = [∑w(|Fo| - |Fc|)2/∑w |Fo|2]1/2 where w is an appropriate weight function based on errors in observed intensities. Typically, all non-H atoms were anisotropically refined and all H atoms except those bound to N and O atoms were calculated by geometric methods and refined using a riding model. X-Ray Powder Diffractometry
[000355] X-ray powder diffraction (PXRD) data were obtained using a manual Bruker GA-DDS (General Area Detector Diffraction System) chi-platform goniometer. Powder samples were placed in 0.7 mm diameter thin-walled glass capillaries; capillaries were rotated during data collection. The distance from the sample to the detector was kept at 17 cm. Data were collected with Cu Kα radiation (À = 1.5418 A) in the range of 2.5 < 2θ < 35° with a sample exposure time of 600 seconds.

权利要求:
Claims (16)
[0001]
1. Compound, characterized by the fact that it has Formula (I):
[0002]
2. Compound according to claim 1, characterized in that: R1 is -CH2CF3 or -CH2CH2CF3; and R2 is -CH2CF3 or -CH2CH2CF3.
[0003]
3. Compound according to claim 1, characterized in that: z is zero or 1.
[0004]
4. Compound according to claim 1, characterized in that: R1 is -CH2CH2CF3; and R2 is -CH2CH2CF3.
[0005]
5. Compound according to claim 4, characterized in that: z is zero or 1.
[0006]
6. Compound according to claim 1, characterized in that: z is 1 or 2.
[0007]
7. Compound according to claim 1, characterized in that it is selected from: (2R,3S)-N-((3S)-1-Methyl-2-oxo-5-phenyl-2,3-di- hydro-1H-1,4-benzodiazepin-3-yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide (1); (2R,3S)-N-((3S)-2-Oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3-bis(3, 3,3-trifluoropropyl)succinamide (2); (2R,3S)-N-((3S)-1-Methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2-(2 ,2,2-trifluoroethyl)-3-(3,3,3-trifluoropropyl)succinamide (3); (2R,3S)-N-((3S)-1-Methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-3-(2 ,2,2-trifluoroethyl)-2-(3,3,3-trifluoropropyl)succinamide (4); (2R,3S)-N-((3S)-1-(2H3)Methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2 ,3-bis(3,3,3-trifluoropropyl)succinamide (5); (2R,3S)-N-((3S)-7-chloro-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (6); (2R,3S)-N-((3S)-8-methoxy-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (7); (2R,3S)-N-((3S)-8-fluoro-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (8); (2R,3S)-N-((3S)-7-methoxy-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (9); (2R,3S)-N-((3S)-7-fluoro-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (10); (2R,3S)-N-((3S)-8-chloro-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (11); (2R,3S)-N-((3S)-9-methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3- bis(3,3,3-trifluoropropyl)succinamide (12); (2R,3S)-N-((3S)-8-methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3- bis(3,3,3-trifluoropropyl)succinamide (13); (2R,3S)-N-((3S)-7-methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3- bis(3,3,3-trifluoropropyl)succinamide (14); (2R,3S)-N-((3S)-8-cyano-9-methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)- 2,3-bis(3,3,3-trifluoropropyl)succinamide (15); (2R,3S)-N-((3S)-8,9-dichloro-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2, 3-bis(3,3,3-trifluoropropyl)succinamide (16); (2R,3S)-N-((3S)-9-fluoro-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3- bis(3,3,3-trifluoropropyl)succinamide (17); (2R,3S)-N-((3S)-9-chloro-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-2,3- bis(3,3,3-trifluoropropyl)succinamide (18); (2R,3S)-N-((3S)-2-Oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-3-(4,4,4 -trifluorobutyl)-2-(3,3,3-trifluoropropyl)succinamide (19); (2R,3S)-N1-((3S)-8-Methoxy-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl)-3-(4 ,4,4-trifluorobutyl)-2-(3,3,3-trifluoropropyl)succinamide (20); and (2R,3S)-N-((3S)-9-((2-Methoxyethyl)amino)-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3 -yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide (21).
[0008]
8. Compound according to claim 1, characterized in that R3 is H or -CH3.
[0009]
9. Compound according to claim 8, characterized in that it is selected from:
[0010]
10. Compound according to claim 9, characterized by the fact that it is:
[0011]
11. A compound according to claim 10, characterized in that said compound is in crystalline Form N-1, which is defined by one of the following: a) a simulated X-ray powder diffraction pattern sub- substantially as shown in Figure 1 and/or by a PXRD pattern substantially observed as shown in Figure 1; b) an X-ray powder diffraction pattern preferably comprising five or more, 2θ values selected from: 5.7±0.2, 20°C; c) unit cell parameters substantially the same as the following: Cell dimensions: a = 9.41 A b = 17.74 A c = 31.94 A α = 90.0° β = 98.4° Y = 90.0 ° Space group: P21 Molecules of said compound/asymmetric unit: wherein the unit cell parameters of Form N-1 are measured at a temperature of about -10°C; and/or d) fractional atomic coordinates substantially as listed in Table 1 at a temperature of about 25°C, Table 1
[0012]
12. Pharmaceutical composition, characterized in that it comprises a compound as defined in any one of claims 1 to 11; and a pharmaceutically acceptable carrier.
[0013]
A compound according to any one of claims 1 to 11, characterized in that it is for use in therapy.
[0014]
A compound according to any one of claims 1 to 11, characterized in that it is for use in the treatment of cancer.
[0015]
15. Combination, characterized in that it comprises a compound as defined in any one of claims 1 to 11; and one or more adding agents selected from dasatinib, paclitaxel, tamoxifen, dexamethasone, and carboplatin.
[0016]
16. Use of a compound, as defined in any one of claims 1 to 11, characterized in that it is in the preparation of a pharmaceutical composition for the treatment of cancer.

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RS53843B1|2015-08-31|

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法律状态:
2018-01-16| B07D| Technical examination (opinion) related to article 229 of industrial property law [chapter 7.4 patent gazette]|

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2018-04-03| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|

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2019-03-19| 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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2019-07-16| B06T| Formal requirements before examination [chapter 6.20 patent gazette]|

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2021-07-06| B06A| Patent application procedure suspended [chapter 6.1 patent gazette]|

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2021-11-09| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2022-01-25| 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 22/03/2012, 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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US201161466238P| true| 2011-03-22|2011-03-22|

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US61/466,238|2011-03-22|

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PCT/US2012/030021|WO2012129353A1|2011-03-22|2012-03-22|Bis-1,4-benzodiazepinone compounds|

---