Methods for Inhibiting MRP1

专利摘要:
The present invention relates to compounds of formula (I) useful for the inhibition of resistant tumors in which resistance is conferred in part or wholly by MRP1. <Formula I>
公开号:KR20010042482A
申请号:KR1020007011102
申请日:1999-04-07
公开日:2001-05-25
发明作者:조셉 마이클 그루버；브라이언 에이치. 노만
申请人:피터 지. 스트링거；일라이 릴리 앤드 캄파니；
IPC主号:

专利说明:

Inhibition method of MRP1 {Methods for Inhibiting MRP1}
Along with surgery and radiotherapy, chemotherapy continues to be an effective treatment for many cancers. In fact, some types of cancer are now considered to be curable by chemotherapy, including Hodgkin's disease, large cell lymphoma, acute lymphocytic leukemia, testicular cancer and early stage breast cancer. Other cancers, such as ovarian cancer, small cell lung and advanced breast cancer, have yet to be cured but have shown a positive response to combination chemotherapy.
One of the most important problems that have not been solved in the treatment of cancer is drug resistance. After selecting resistance to a single cytotoxic drug, the cells become cross resistant to a full range of drugs with different structures and cellular targets, such as alkylating agents, antimetabolic, hormones, platinum-containing drugs and natural products. Can be. This phenomenon is known as multidrug resistance (MDR). In some types of cells, this resistance is innate, while in others, for example, small cell lung cancer is generally acquired.
This resistance is known to be multifactorial and is conferred by at least two proteins: 170 kDa P-glycoprotein (MDR1) and the more recently identified 190 kDa multidrug resistance protein (MRP1). Although both MDR1 and MRP1 belong to the ATP-binding cassette superfamily of transport proteins, they are structurally very different molecules and share less than 15% amino acid homology. Despite the structural differences between the two proteins, no consistent difference was known between MDR1 and MRP1 cell line resistance patterns until 1994. However, association of MRP1 or its deficiency and resistance to certain oncolytics have been known [see, eg, Cole, et.al., "Pharmacological Characterization of Multidrug Resistant MRP-transfected Human Tumor Cells", Cancer Research, 54: 5902-5910, 1994]. Doxorubicin, daunorubicin, epirubicin, vincristine and etoposide are substrates of MRP1, ie, MRP1 can bind to these tumor cell disruptors and redistribute them away from their site of action, nucleus and cells [Id . and Marquardt, D., and Center, M.S., Cancer Research, 52: 3157, 1992].
Doxorubicin, daunorubicin and epirubicin are members of anthracycline-based tumor cell disruptors. These act as isolates of various strains of Streptomyces by inhibiting nucleic acid synthesis. These agents are useful for treating bone, ovary, bladder, thyroid, and especially neoplasms of the breast. They are also useful for the treatment of acute lymphoblastic and myeloid leukemia, Wilms' tumors, neuroblastomas, soft tissue sarcomas, Hodgkin's and non-Hodgkin's lymphomas, and organ supportive cancers.
Vincristine, a member of the vinca alkaloid-based tumor cell disintegrator, is a isolate of common flowering plants, Vinca rosea Linn. The mechanism of action of vincristine is still under study but has been associated with the inhibition of microtubule formation in mitotic spindles. Vincristine is useful for the treatment of acute leukemia, Hodgkin's disease, non-Hodgkin's malignant lymphoma, rhabdomyosarcoma, neuroblastoma and Wilms' tumor.
Etoposide, a member of the epipodophyllotoxin-based tumor cell disruptor, is a semisynthetic derivative of grapephytotoxin. Etoposide acts as a topoisomerase inhibitor and is useful for tumor therapy in the testes and lungs.
It is currently unknown what determines the cell line to acquire resistance through the MDR1 or MRP1 mechanism. Due to the tissue specificity of these transporters and / or if one mechanism prevails or there is only one mechanism, it is useful to have its selective inhibitors over the other. In addition, when administering a drug or drugs that are substrates of either protein, it may be particularly advantageous to co-administer the drug that is a selective inhibitor of that protein. Therefore, it is desirable to provide compounds that are selective inhibitors of MDR1 or MRP1.
The present invention relates to compounds of formula (I) or pharmaceutical salts or solvates thereof.
Where
m is an integer from 1 to 6,
R is COR ¹ , amino, NH-Pg or NHCOR ² ,
R ¹ is hydroxy, C ₁ -C ₆ alkoxy, or NR ³ R ⁴ ,
Pg is an amino protecting group,
R ² is C ₁ -C ₆ alkyl, substituted C ₁ -C ₄ alkyl, aryl, substituted aryl, (CH ₂ ) _n -heterocycle, (CH ₂ ) _n -substituted heterocycle,
R ³ is independently at each occurrence hydrogen or C ₁ -C ₆ alkyl,
R ⁴ is C ₁ -C ₆ alkyl, norbornan-2-yl, aryl, substituted aryl, CH ₂ CH (CH ₃ ) phenyl, (CH ₂ ) _n heterocycle or (CH ₂ ) _n -substituted hetero Cycle,
n is 0, 1 or 2.
The present invention also relates to a method of inhibiting MRP1 in a mammal comprising administering an effective amount of a compound of formula (I) or a pharmaceutical salt or solvate thereof to a mammal in need of inhibition of MRP1.
In another embodiment, the invention provides an effective amount of a compound of Formula (I) or a pharmaceutical salt or solvate thereof in combination with an effective amount of tumor cell disintegrator to administer to a mammal in need of inhibition of a resistant tumor or a tumor that is likely to be resistant. It relates to a method for inhibiting a resistant tumor or a tumor that is likely to be resistant in a mammal.
The invention also relates to a pharmaceutical formulation comprising a compound of formula (I) or a pharmaceutical salt or solvate thereof in combination with one or more tumor cell disintegrating agents, pharmaceutical carriers, diluents or excipients.
The present invention is a compound of the selected group, i.e., a compound of Formula I, is a selective inhibitor of multidrug resistant protein (MRP1) and is therefore useful for treating MRPl conferred multidrug resistance (MDR) in resistant tumors and tumors that are prone to resistance. .
The terms "inhibit" and "inhibit MRP1" when associated with MRP1 prevent, moderate, ameliorate, stop, suppress, slow or reverse the progression of MRP1, or site of tumor cell disinfectant action, most It is usually referred to as reducing the ability of MRP1 to redistribute tumor cell disintegrating agents away from the nucleus and away from the tumor.
As used herein, the term “effective amount of a compound of formula (I)” refers to an amount of a compound of the present invention that can inhibit MRP1. The term "effective amount of tumor cell disruptor" refers to the amount of tumor cell disruptor that can inhibit the tumor in a resistant or otherwise manner.
The term "inhibits a resistant tumor, or a tumor that is susceptible to resistance" prevents, alleviates, ameliorates, stops, suppresses or reverses the progression of a resistant tumor and / or a tumor that is susceptible to resistance, or is resistant Reducing the growth of tumors and / or susceptible tumors or killing resistant and / or susceptible tumors.
The term "resistant tumor" refers to a tumor that is resistant to chemotherapy in which resistance is conferred in part or entirely by MRP1. Such tumors include, but are not limited to, tumors of the bladder, bone, breast, lung (small cells), testicles, and thyroid gland, and also include, but are not limited to, acute lymphoblastic and myeloid leukemia, Wilms' tumor, neuroblastoma, and soft tissue. More specific types of cancers such as, but not limited to, sarcomas, Hodgkin's and non-Hodgkin's lymphomas, and organ supportive cancers.
A “prone to resistant” tumor is a tumor that is neither innate nor currently present, but can be conferred by MRP1 after chemotherapy has begun. Thus, the methods of the present invention include prophylactic and therapeutic administration of a compound of formula (I).
The term “chemotherapy” refers to the use of at least one tumor cell disruptor wherein at least one tumor cell disruptor is a substrate of MRP1. A "substrate of MRP1" is a tumor cell disruptor that binds to MRP1 and is redistributed away from the site of action (the nucleus of the tumor) and away from the cell, thus making therapy less effective.
The term “treat” or “treatment” prevents, prevents, alleviates, improves, stops, suppresses, slows down or reverses the progression of MRP1 induced drug resistance in a multidrug resistant tumor, or the depth of MRP1 induced drug resistance. Has their general meaning, including reducing it.
In the formulas herein, general chemical terms have their usual meanings. For example, the term “C ₁ -C ₄ alkyl” refers to methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, cyclopentyl, s-butyl and t-butyl. The term “C ₁ -C ₆ alkyl” refers to a monovalent straight chain, branched or cyclic saturated hydrocarbon containing 1 to 6 carbon atoms. In addition, C ₁ -C ₆ alkyl also includes, but is not limited to, cyclopentyl, pentyl, hexyl, cyclohexyl, and the like.
The term "aryl" refers to phenyl, benzyl and naphthyl.
The term "heterocycle" refers to a monovalent saturated, unsaturated or aromatic monocyclic or fused ring having from 5 to 7 or 8 to 10 total atoms each, containing 1 to 3 heteroatoms independently selected from oxygen, sulfur and nitrogen. Say system. Examples of heterocyclic groups include furanyl, indolyl, thiophenyl, isoxazolyl, all of their partially saturated or fully saturated analogs, for example rings in which the point of attachment to the hetero cycle can be used for substitution Tetrahydrofuran and the like at any position of the phase, but are not limited thereto.
The term “substituted heterocycle” independently refers to a heterocycle ring substituted one or two times with a C ₁ -C ₆ alkyl, halo, benzyl, phenyl, trifluoromethyl or oxo group.
The term “substituted aryl” is each independently halo, hydroxy, trifluoromethyl, N (R ¹ ) ₂ , NH-Pg, C ₁ -C ₄ alkoxy, benzyloxy, CO ₂ R ¹ , SO ₂ NH ₂ , Phenyl, benzyl, and naphthyl groups substituted one or five times with trifluoromethoxy or nitro.
The term “substituted C ₁ -C ₄ alkyl” refers to a C ₁ -C ₄ alkyl group, eg trifluoromethyl, in which one to three hydrogens are replaced with the same halide.
The term "halo" or "halide" refers to fluoro, chloro, bromo or iodo.
The term "C ₁ -C ₄ alkoxy" refers to a C ₁ -C ₄ alkyl group bonded via an oxygen atom.
As used herein, the term “amino protecting group” refers to the substituent (s) of the amino group that is generally used to block or protect the amino function while reacting with other functional groups on the compound. Examples of the amino-protecting group include formyl group, trityl group, phthalimido group, acetyl group, trichloroacetyl group, chloroacetyl group, bromoacetyl group, and iodoacetyl group, urethane-type blocking group, for example And amino protecting groups such as benzyloxycarbonyl and 9-fluorenylmethoxycarbonyl ("FMOC"). The type of amino protecting group used is not critical as long as the derived amino group is stable to the conditions of subsequent reaction (s) on different positions of the molecule and can be removed at an appropriate time without destroying the rest of the molecule. Similar amino protecting groups used in the field of cephalosporins, penicillins and peptides are also encompassed by the above terms. Further examples of groups referred to by the above terms are described in T.W. Greene, "Protective Groups in Organic Synthesis", John Wiley and Sons, New York, N.Y., 1991, Chapter 7]. Preferred amino protecting groups are t-butyloxycarbonyl.
The term "carbonyl activator" refers to a substituent of carbonyl that makes the carbonyl group susceptible to nucleophilic addition. Suitable activators are those having a net electron attraction effect on carbonyl. Such groups include alkoxy, aryloxy, nitrogen containing aromatic heterocycles, or amino groups such as oxybenzotriazole, imidazolyl, nitrophenoxy, pentachlorophenoxy, N-oxysuccinimide, N, N'- Dicyclohexylisoure-O-yl, N-hydroxy-N-methoxyamino and the like; Acetates, formates, sulfonates such as methanesulfonate, ethanesulfonate, benzenesulfonate or p-toluenylsulfonate and the like; And halides, especially chloride, bromide or iodide, but are not limited to these.
In general, when used as an adjective, the term "pharmaceutical" means substantially nontoxic to living organisms. For example, the term "pharmaceutical salt" as used herein refers to salts of compounds of formula I that are substantially nontoxic to living organisms [eg, Berge, SM, Bighley, LD, and Monkhouse, DC, " Pharmaceutical Salts ", J. Pharm. Sci., 66: 1, 1977]. Representative pharmaceutical salts include salts prepared by reacting a compound of formula I with an inorganic or organic acid or base. These salts are known as acid addition salts or base addition salts, respectively. These pharmaceutical salts mainly have improved solubility properties as compared to the compounds from which they are derived, and therefore are often more easily formulated as ligands or emulsions.
Examples of pharmaceutical acid addition salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromide, Iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, succinate, suverate, Sebacate, fumarate, maleate, butyne-1,4-dioate, hexyn-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxy Benzoate, phthalate, sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citre Agent, such as a lactate, γ--hydroxybutyrate, glycolate, tartrate, methanesulfonate, ethanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate rate.
Examples of pharmaceutical base addition salts are the ammonium, lithium, potassium, sodium, calcium, magnesium, methylamino, diethylamino, ethylene diamino, cyclohexylamino, ethanolamino salts and the like of the compounds of formula (I).
The term “solvate” refers to an aggregate comprising at least one solute molecule and at least one solvent molecule, such as a compound of formula (I).
The term "suitable solvent" refers to a solvent that is inert to the ongoing reaction and sufficiently dissolves the reactants to effect the desired reaction. Examples of suitable solvents include dichloromethane, chloroform, 1,2-dichloroethane, diethyl ether, acetonitrile, ethyl acetate, 1,3-dimethyl-2-imidazolidinone, tetrahydrofuran, dimethylformamide, toluene, chloro Benzene, dimethyl sulfoxide, mixtures thereof, and the like, but are not limited thereto.
The term "carbonyl activator" refers to a reagent that converts a carbonyl group of a carboxylic acid into a more nucleophilic addition reaction, referred to as "The Peptides" and "Peptide Synthesis", respectively, "The Peptides", Gross and Meienhofer, Eds., Academic Press (1979), Ch. 2 and M Bodanszky, "Principles of Peptide Synthesis :, 2 ^nd Ed., Springer-Verlag Berlin Heidelberg, 1993", including but not limited to. Specifically, carbonyl activators include Nucleophiles of hydrogen such as thionyl bromide, thionyl chloride, oxalyl chloride and the like, alcohols such as nitrophenols, pentachlorophenol and the like, amines such as N-hydroxy-N-methoxyamine and the like; Acid halides such as acetic acid, formic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid or p-toluenesulfonic acid halides and the like; compounds such as 1,1'-carbonyldiimidazole, benzotriazole, imidazole, N -Hydroxysuccinimide, dicyclohexylcarbodiimide, etc. are mentioned.
The term “suitable thermodynamic base” is sufficiently reactive and acidic to effect the desired reaction without significantly effecting any undesirable reaction or base which acts as a proton trap for any proton that can be produced as a byproduct of the desired reaction. Refers to a base that provides for reversible deprotonation of a substrate. Examples of thermodynamic bases include carbonates, bicarbonates and hydroxides (eg carbonic acid, bicarbonate or lithium hydroxide, sodium or potassium), tri- (C ₁ -C ₄ alkyl) amines or aromatic nitrogen containing heterocycles (eg pyridine) Although it is mentioned, it is not limited to these.
While all compounds of the present invention are useful, certain compounds are particularly advantageous and preferred. The following list lists several groups of preferred compounds, agents, and methods. Each of these may be combined with another to create a further group of preferred embodiments.
a) m is an integer of 2-6.
b) R is amino
c) R is t-butyloxycarbonylamino.
d) R is trifluoroacetylamino.
e) R is 3,4,5-trimethoxybenzoylamino.
f) R is carboxy.
g) R is 3,4,5-trimethoxyanilinylcarboxy.
h) R is 3,4,5-trimethoxybenzylamiminylcarboxy.
i) a compound of the example part.
j) a compound that is a pharmaceutical salt.
k) A compound that is a hydrochloride salt.
l) The mammal is a human.
m) A method wherein the tumor cell disintegrant is selected from doxorubicin, daunorubicin, epirubicin, vincristine and etoposide.
n) the tumor is of Wilms type, bladder, bone, breast, lung (small cell), testicular or thyroid or the tumor is acute lymphoblastic and myeloid leukemia, neuroblastoma, soft tissue sarcoma, Hodgkin's and non-Hodgkin's lymphoma , Or a method associated with organ support cancer. And
o) An agent wherein the tumor cell disintegrating agent is selected from doxorubicin, daunorubicin, epirubicin, vincristine and etoposide.
The compounds of the present invention can be prepared by a variety of methods, some of which are illustrated in the following schemes. The specific order of steps necessary to prepare the compound of formula I depends on the relative instability of the particular compound, derivatized compound and substituted group to be synthesized.
Compounds of formula (I) may be prepared from compounds of formula (II) as R and m are illustrated in Scheme 1 below, as described above.
Compounds of formula (I) can be prepared by dissolving or suspending the compound of formula (II) in a suitable solvent and adding a suitable thermodynamic base. Typically, a preferred and convenient solvent is dimethylformamide. In general, a convenient and preferred thermodynamic base is sodium hydroxide added as a 2N solution in methanol. The reactions are typically combined at room temperature and the resulting solution is typically reacted for 30 minutes to about 18 hours. Preferably, the mixture is stirred for about 1 to about 10 hours, most preferably for about 3.5 hours to about 7 hours. Bases are typically used in large molar excesses, and generally from about 4 to about 8 molar excesses relative to the compound of formula II. Preferably, about 5 to about 7 molar excess is typically used. Certain intermediates of the compounds of formula (I) discussed below may also be prepared by the methods described above.
Any hydroxy or amino protecting group found in the cyclized compound of formula (I) may optionally be removed as described for Greene to provide the free amino or free hydroxy compound of formula (I). Preferred choices for protecting groups and their removal methods can be found in the Preparations and Examples section below.
Compounds of formula (I) wherein R is COR ¹ also include R ⁵ being a carboxy activator, R ⁶ being C ₁ -C ₆ alkoxy or NR ³ R ⁴ , and m as illustrated in Scheme 2 below: It may also be prepared from a compound of formula I (a).
Compounds of formula (I) prepared as described in Scheme 1 may be converted to other compounds of the invention. For example, the activated carboxylic acid of formula (III) may be formed by methods known in the chemical art by activating the acid of formula (I) [eg, The Peptide, Peptide Synthesis, and See examples and preparation examples below.
The compound of formula I (b) can then be prepared by dissolving or suspending the compound of formula III in a suitable solvent, optionally in the presence of a suitable thermodynamic base, and adding the amine of formula IV. Typically, a preferred and convenient solvent is dichloromethane. Preferred bases are triethylamine and piperidinylmethylpolystyrene resins. Amines are typically used in molar excess. For example, about 1.5 to about 3 molar excess is generally used for the compound of Formula III. Typically from about 1.8 to about 2.2 molar excess is preferred. The reaction is generally carried out for about 10 minutes to about 18 hours in the temperature range of about 0 ° C to the reflux temperature of the solvent. Preferably, the reaction is carried out at about 15 ° C to about 40 ° C for 5 minutes to about 2.5 hours.
Alternatively, the compound of formula (I) can be activated and the addition of the compound of formula (IV) can be carried out in one pot process as described in Examples 26 and 38 below.
Compounds of formula (I), ie esters, wherein R ⁶ is C ₁ -C ₆ alkoxy, can be prepared by methods well known in the chemical art. For instructions on the conversion of activated carboxylic acids to esters, see, eg, Larock, "Comprehensive Organic Transgormations", pgs. 978-979, VCH Publishers, New York, NY, 1989. Alternatively, these compounds of formula I (b) may be prepared directly from the acids of formula I (a) as described on Larock pages 966-972.
Compounds of formula I wherein R is amino or NHCOR ² can be prepared from compounds of formula I (c) as m Pg, R ² and R ⁵ are illustrated in Scheme 3 below as described above.
Compounds of formula (I) may be converted to other compounds of the invention. For example, a compound of formula (I) may have Greene and its protecting group removed as described in the Examples section below, which may then form a compound of formula (I). These compounds of formula I (d) can then also be converted to other compounds of the invention. For example, a compound of formula (I) may be treated with a compound of formula (VI), optionally dissolved or suspended in a suitable solvent in the presence of a thermodynamic base to provide a compound of formula (I). Typically, a preferred and convenient solvent is dimethylformamide or a mixture of dichloromethane and dimethylformamide. When a base is used, triethylamine or N-methylmorpholine is typically the preferred base. In addition, when a base is used, the base and the compound of formula VI are typically used in stoichiometric to high molar excesses. For example, an excess of 1.0 to 4 moles of the compound of formula (I) is generally used. When no base is used, the compound of formula IV is typically used in relatively more stoichiometric excess. Preferably, the reaction is carried out in the presence of a base with about 1.8 to about 2.2 equivalents of the compound of formula VI. The reaction is generally carried out for 10 minutes to 24 hours in the temperature range of about 0 ° C. to about the reflux temperature of the solvent. Preferably, the reaction is carried out at about 15 ° C to about 40 ° C for 4 to about 18 hours.
Pharmaceutical salts of the present invention are typically formed by reacting an equivalent or excess acid or base with a compound of formula (I). The reaction is generally mixed in mutual solvents such as diethyl ether, tetrahydrofuran, methanol, ethanol, isopropanol, benzene, etc. for acid addition salts, or in water, alcohol or chlorinated solvents, for example dichloromethane for base addition salts. The salts precipitate out of solution normally within about 1 hour to about 10 days and can be isolated by filtration or other conventional methods.
Acids commonly used to produce pharmaceutical acid addition salts are inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, and the like, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, oxalic acid, p-bromophenylsulfonic acid, carboxylic acid , Organic acids such as succinic acid, citric acid, tartaric acid, benzoic acid, acetic acid and the like. Preferred pharmaceutical acid addition salts are formed with inorganic acids such as hydrochloric acid, hydrobromic acid and sulfuric acid and are formed with organic acids such as maleic acid, tartaric acid and methanesulfonic acid.
Commonly used bases for preparing pharmaceutical base addition salts are inorganic bases such as ammonium or alkali or alkaline earth metal hydroxides, carbonates, bicarbonates and the like. Thus, such bases useful for preparing the salts of the present invention include sodium hydroxide, potassium hydroxide, ammonium hydroxide, potassium carbonate, sodium carbonate, sodium bicarbonate, potassium bicarbonate, calcium hydroxide, calcium carbonate and the like. Particular preference is given to potassium and sodium salt forms.
It should be understood that the particular counterion that forms part of any salt of the present invention has no important properties unless the salt is generally pharmacologically acceptable and the counterion generally provides the salt with undesirable quality.
Starting materials of the methods of the invention can be obtained by a number of routes. For example, compounds of Formulas II, I (a), and I (c) can be prepared according to the route in which m and R are shown in Scheme 4 below, as described above.
Compounds of formula (II) can be prepared by dissolving or suspending the compound of formula (VII) in a suitable solvent and adding the compound of formula (VIII) and a suitable thermodynamic base. Dichloromethane is typically preferred as a convenient solvent. Triethylamine is generally the preferred thermodynamic base. This amide formation reaction is also preferably carried out in the presence of diethylamino pyridine (DMAP). Compounds of formula (VIII) are typically and preferably used in equimolar amounts relative to compounds of formula (VII), although some excess (about 0.05 to about 0.15 molar excess) is also acceptable. Thermodynamic bases are typically used in slightly molar excesses, for example, from about 1.01 to about 1.2 molar excesses are typically used for compounds of Formula (VII). An about 1.05 to about 1.15 molar excess is generally preferred. DMAP is used in a catalytic manner. For example, about 5 mol% to about 15 mol% are typically used relative to the compound of formula VII. 10 mole% is generally preferred.
Although the transformations described in Schemes 2 and 3 can be carried out prior to the cyclization described in Scheme 1, it is preferred to carry out these reactions after cyclization with the R substituents fully described. Accordingly, preferred starting materials for the reactions of Schemes 1 and 4 are compounds of Formulas II and VII wherein R is NH-Pg or COR ¹ , wherein R ¹ is C ₁ -C ₆ alkoxy. In addition, when the reaction of Scheme 1 is carried out with a compound of the formula II wherein R is COR ¹ and R ¹ is C ₁ -C ₆ alkoxy, under the conditions described for the cyclization, the ester is decomposed to acid That is, to produce a compound of formula (I).
Compounds of formulas (IV), (V), (VI), (VII) and (VIII) are well known in the art and are not readily commercially available, but are readily synthesized by standard methods commonly used in the art.
The optimal time for carrying out the reactions of Schemes 1-4 can be determined by monitoring the progress of the reaction via conventional chromatography techniques. It is also preferred to carry out the reaction of the invention under an inert atmosphere, for example argon or in particular nitrogen. The choice of solvent is generally not critical, as long as the solvent used is inert to the ongoing reaction and can sufficiently dissolve the reactants to enable the desired reaction. Compounds of formula (I) and (II) are preferably isolated and purified before being used for their subsequent reactions. These compounds can be crystallized from the reaction solution while they are formed and then collected by filtration, or the reaction solvent can be extracted, evaporated or decanted and thus removed. These intermediates of formula (I) and final products may be further purified by general techniques, for example by recrystallization or by chromatography on a solid support, such as silica gel or alumina, if necessary.
The following formulation examples and examples are provided to better illustrate the practice of the invention and should not be construed as limiting the scope of the invention in any sense. Those skilled in the art will recognize that various modifications may be made without departing from the spirit and scope of the invention. All publications mentioned in the specification are indicative of the level of ordinary skill in the art to which this invention pertains. The terms and abbreviations used in the preparations and examples have their usual meanings unless stated otherwise. For example, "° C.", "N", "mmol", "g", "ml", "M", "IR", "MS (FD)" and "MS (IS)" are each Celsius, Normal or normal concentration, millimolar, gram, milliliter, molar or molar concentration, infrared spectroscopy, field emission mass spectroscopy, and ion spray mass spectroscopy. Also, the absorption maximums listed for the IR spectrum are not all maximums observed to be important.
<Production example>
<Production example 1>
N-t-butyloxycarbonyl-N '-(3- [2-chloro-6-fluorophenyl] -5-methylisoxazol-4-oil) -1,2-diaminoethane
355 mg (2.22 mmol) of N-t-butyloxycarbonyl-1,2-diaminoethane, 27 mg (0.222 mmol) of dimethylaminopyridine and 247 mg (2.44 mmol) of triethylamine were combined in 5 mL of anhydrous dichloromethane. 607 mg (2.22 mmol) of 3- (2-chloro-6-fluorophenyl) -5-methylisoxazole-4-oil chloride were added slowly. The reaction was stirred at room temperature under nitrogen for 3 hours. The reaction was diluted with ethyl acetate and water. Ethyl acetate was separated and washed three times with 25 ml of 1N aqueous hydrochloric acid solution and brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was treated with hexane to crystallize 794 mg of the title compound (90%).
Elemental Analysis for C ₈ H ₂₁ N ₃ O ₄ ClF (EA):
Calc .: C, 54.34; H, 5. 32; N, 10.56.
Found: C, 54.30; H, 5. 36; N, 10.44.
MS (IS) m / z 398 (M &lt; + &gt;).
<Production example 2>
N-t-butyloxycarbonyl-N '-(3- [2-chloro-6-fluorophenyl] -5-methylisoxazol-4-oil) -1,3-diaminopropane
574 mg (3.29 mmol) of Nt-butyloxycarbonyl-1,3-diaminopropane and 903 mg (3.29 mmol) of 3- (2-chloro-6-fluorophenyl) -5-methylisoxazole-4-oil chloride ) Was converted to 1.36 g of the title compound by the method of Preparation Example 1 (100%).
Elemental Analysis for C ₁₉ H ₂₃ N ₃ O ₄ ClF (EA):
Calc .: C, 55.41; H, 5.63; N, 10.20.
Found: C, 55.17; H, 5.71; N, 9.99.
MS (IS) m / z 412 (M &lt; + &gt;).
<Production example 3>
N-t-butyloxycarbonyl-N '-(3- [2-chloro-6-fluorophenyl] -5-methylisoxazol-4-yl) -1,4-diaminobutane
985 mg (5.23 mmol) of Nt-butyloxycarbonyl-1,4-diaminopropane and 1.43 g (5.23 mmol) of 3- (2-chloro-6-fluorophenyl) -5-methylisoxazole-4-yl chloride ) Was converted to 2.23 g of the title compound by the method of Preparation Example 1 except that the reaction time was about 18 hours (100%).
Elemental Analysis for C ₂₀ H ₂₅ N ₃ O ₄ ClF (EA):
Calc .: C, 56.41; H, 5.92; N, 9.87.
Found: C, 56.12; H, 5.75; N, 9.67.
MS (FD) m / z 426 (M &lt; + &gt;).
<Production example 4>
N-t-butyloxycarbonyl-N '-(3- [2-chloro-6-fluorophenyl] -5-methylisoxazol-4-yl) -1,5-diaminopentane
1.04 g (5.14 mmol) of Nt-butyloxycarbonyl-1,5-diaminopentane and 1.41 g (5.14 mmol) of 3- (2-chloro-6-fluorophenyl) -5-methylisoxazole-4-oil chloride ) Was converted to 2.14 g of the title compound by the method of Preparation Example 1 (95%).
MS (IS) m / z 412 (M &lt; + &gt;).
IR (CHCl ₃ ) 3496, 3012, 2936, 1708, 1663, 1611, 1510 cm ⁻¹ .
<Production example 5>
N-t-butyloxycarbonyl-N '-(3- [2-chloro-6-fluorophenyl] -5-methylisoxazole-4-yl) -1,6-diaminohexane
1.02 g (4.03 mmol) of Nt-butyloxycarbonyl-1,6-diaminohexane hydrochloride salt and 1.11 g of 3- (2-chloro-6-fluorophenyl) -5-methylisoxazole-4-yl chloride (4.03 mmol) was converted to 1.84 g of the title compound by the method of Preparation Example 1 (100%).
Elemental Analysis for C ₂₂ H ₂₉ N ₃ O ₄ ClF (EA):
Calc .: C, 58.21; H, 6. 44; N, 9.26.
Found: C, 57.97; H, 6. 20; N, 9.43.
MS (IS) m / z 454 (M &lt; + &gt;).
<Production example 6>
Methyl-4-aminobutanoate hydrochloride
1 g (4.92 mmol) of N-t-butyloxycarbonyl-4-aminobutanoic acid was dissolved in 10 ml of a freshly prepared mixture of hydrochloric acid and methanol (2.5 ml of acetyl chloride in 35 ml of methanol). The reaction was stirred for about 2 hours and then stripped. The residue was dissolved in ethyl acetate, washed three times with aqueous sodium bicarbonate solution and brine, respectively, dried over sodium sulfate, filtered and concentrated to give 708 mg (94%) of the title compound.
Elemental Analysis for C ₅ H ₁₂ ClNO ₂ (EA):
Calc .: C, 39.10; H, 7.87; N, 9.12.
Found: C, 38.89; H, 7.65; N, 8.94.
MS (FD) m / z 118 (M &lt; + &gt; for free amines).
<Production example 7>
Methyl-5-aminopentanoate hydrochloride
1 g (4.60 mmol) of N-t-butyloxycarbonyl-5-aminopentanoic acid was converted to the title compound by the method of Preparation 6 to give 705 mg (91%).
Elemental Analysis for C ₆ H ₁₄ ClNO ₂ (EA):
Calc .: C, 42.99; H, 8.42; N, 8.36.
Found: C, 43.02; H, 8. 15; N, 8.18.
MS (FD) m / z 132 (M &lt; + &gt; for free amines).
<Production example 8>
Methyl-6-aminohexanoate hydrochloride
500 mg (2.16 mmol) of N-t-butyloxycarbonyl-6-aminohexanoic acid were converted to the title compound by the method of Preparation Example 6 to give 357 mg (91%).
Elemental Analysis for C ₇ H ₁₆ ClNO ₂ (EA):
Calc .: C, 46.28; H, 8.88; N, 7.71.
Found: C, 46.04; H, 8.88; N, 7.51.
MS (FD) m / z 146 (M &lt; + &gt; for free amines).
<Production example 9>
Methyl-7-aminoheptanoate hydrochloride
1 g (4.08 mmol) of N-t-butyloxycarbonyl-7-aminoheptanoic acid was converted to the title compound by the method of Preparation 6 to give 742 mg (93%).
Elemental Analysis for C ₈ H ₁₇ ClNO ₂ (EA):
Calc .: C, 49.10; H, 9. 27; N, 7.16.
Found: C, 49.10; H, 9. 18; N, 7.03.
MS (FD) m / z 160 (M &lt; + &gt; for free amines).
<Production example 10>
Methyl-N- (3- [2-chloro-6-fluorophenyl] -5-methylisoxazole-4-oyl) -4-aminobutanoate
668 mg (4.34 mmol) and 3- (2-chloro-6-fluorophenyl) -methyl-4-aminobutanoate hydrochloride by the method of Preparation Example 1, except that the residue was not treated with hexane. 1.19 g (4.35 mmol) of 5-methylisoxazole-4-oyl chloride were converted to 1.60 g of the title compound (100%).
Elemental Analysis for C ₁₆ H ₁₆ N ₂ O ₄ ClF:
Calc .: C, 54.17; H, 4.55; N, 7.90.
Found: C, 54.41; H, 4.58; N, 7.78.
MS (FD) m / z 355 (M &lt; + &gt;).
<Production example 11>
Methyl-N- (3- [2-chloro-6-fluorophenyl] -5-methylisoxazole-4-oil) -5-aminopentanoate
694 mg (4.14 mmol) of methyl-5-aminopentanoate hydrochloride and 1.24 g of 3- (2-chloro-6-fluorophenyl) -5-methylisoxazole-4-yl chloride by the method of Preparation Example 10 4.52 mmol) was converted to 1.36 g of the title compound (100%).
Elemental Analysis for C ₁₇ H ₁₈ N ₂ O ₄ ClF (EA):
Calc .: C, 55.37; H, 4.92; N, 7.60.
Found: C, 55.44; H, 5.00; N, 7.52.
MS (FD) m / z 368 (M &lt; + &gt;).
<Production example 12>
Methyl-N- (3- [2-chloro-6-fluorophenyl] -5-methylisoxazole-4-oyl) -6-aminohexanoate
349 mg (1.92 mmol) of methyl-6-aminohexanoate hydrochloride and 527 mg of 3- (2-chloro-6-fluorophenyl) -5-methylisoxazole-4-yl chloride by the method of Preparation Example 10 (1.92 mmol) was converted to 2.23 g of the title compound (100%).
Elemental Analysis for C ₁₈ H ₂₀ N ₂ O ₄ ClF (EA):
Calc .: C, 56.48; H, 5. 27; N, 7.32.
Found: C, 56.66; H, 5.20; N, 7.07.
MS (FD) m / z 382.2 (M &lt; + &gt;).
<Production example 13>
Methyl-N- (3- [2-chloro-6-fluorophenyl] -5-methylisoxazol-4-oyl) -7-aminoheptanoate
733 mg (3.75 mmol) of methyl-7-aminoheptanoate hydrochloride and 1.03 g of 3- (2-chloro-6-fluorophenyl) -5-methylisoxazole-4-yl chloride by the method of Preparation Example 10 (3.75 mmol) was converted to 1.48 g of the title compound (100%).
Elemental Analysis for C ₁₉ H ₂₂ N ₂ O ₄ ClF (EA):
Calc .: C, 57.51; H, 5.59; N, 7.06.
Found: C, 57.78; H, 5. 70; N, 6.80.
MS (FD) m / z 396.1 (M &lt; + &gt;).
<Example 1>
1- (N-t-butyloxycarbonyl-2-aminoethyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
772 mg (1.94 mmol) of Nt-butyloxycarbonyl-N '-(3- (2-chloro-6-fluorophenyl) -5-methylisoxazole-4-oil) ethylene-1,2-diamine It was dissolved in 10 ml of dimethylformamide and stirred under nitrogen at room temperature. 5 ml of aqueous sodium hydroxide solution was added and the resulting solution was stirred for 4 hours. The reaction was partitioned between 1N aqueous hydrochloric acid solution and 50 ml of ethyl acetate. Ethyl acetate was washed three times with 25 ml brine and dried over sodium sulfate. The solution was concentrated in vacuo to give 721 mg of the title compound (98%).
Elemental Analysis for C ₁₅ H ₂₀ ClN ₃ O ₄ (EA):
Calc .: C, 57.22; H, 5. 34; N, 11.12.
Found: C, 57.21; H, 5. 23; N, 10.97.
MS (IS) m / z 376 (M-H).
<Example 2>
1- (N-t-butyloxycarbonyl-3-aminopropyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
Nt-butyloxycarbonyl-N '-(3- (2-chloro-6-fluorophenyl) -5-methylisoxazole-4 by the method of Example 1 except that the reaction time was 6 hours 1.31 g (3.18 mmol) of -oil) -1,3-diaminopropane were converted to 1.22 g of the title compound (99%).
Elemental Analysis for C ₁₉ H ₂₂ ClN ₃ O ₄ (EA):
Calc .: C, 58.24; H, 5. 66; N, 10.72.
Found: C, 58.48; H, 5.52; N, 10.73.
MS (IS) m / z 392 (M &lt; + &gt;).
<Example 3>
1- (N-t-butyloxycarbonyl-4-aminobutyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
Nt-butyloxycarbonyl-N '-(3- (2-chloro-6-fluorophenyl) by the method of Example 1 except that the residue was azeotropic with xylene after concentration to remove residual dimethylformamide. 2.23 g (5.23 mmol) of) -5-methylisoxazole-4-oil) -1,4-diaminobutane were converted to 2.11 g of the title compound (99%).
Elemental Analysis for C ₂₀ H ₂₄ ClN ₃ O ₄ (EA):
Calc .: C, 59.19; H, 5.96; N, 10.35.
Found: C, 58.71; H, 5.92; N, 9.96.
MS (FD) m / z 405 (M &lt; + &gt;).
<Example 4>
1- (N-t-butyloxycarbonyl-5-aminopentyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
The reaction time was 6 hours, and Nt-butyloxycarbonyl-N '-(3- (2-chloro-6- was obtained by the method of Example 1, except that the residue was recrystallized from dichloromethane: hexane. 2.11 g (4.80 mmol) of fluorophenyl) -5-methylisoxazole-4-oyl) -1,5-diaminopentane were converted to 1.13 g of the title compound (56%).
Elemental Analysis for C ₂₁ H ₂₆ ClN ₃ O ₄ (EA):
Calc .: C, 60.07; H, 6. 24; N, 10.01.
Found: C, 60.07; H, 6. 30; N, 10.11.
MS (IS) m / z 420 (M &lt; + &gt;).
<Example 5>
1- (N-t-butyloxycarbonyl-6-aminohexyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
Nt-butyloxycarbonyl-N '-(3- (2-chloro-6-fluorophenyl) -5-methylisoxazol-4-yl) -1,6-diaminohexane by the method of Example 3 1.84 g (4.05 mmol) was converted to 1.45 g of the title compound (82%).
Elemental Analysis for C ₂₂ H ₂₈ ClN ₃ O ₄ (EA):
Calc .: C, 60.89; H, 6. 50; N, 9.68.
Found: C, 61.19; H, 6. 33; N, 9.51.
MS (IS) m / z 434.6 (M &lt; + &gt;).
<Example 6>
1- (2-aminoethyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one trifluoroacetate
687 mg (1.82 of 1- (Nt-butyloxycarbonyl-2-aminoethyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one mmol) was dissolved in 10 ml of dichloromethane and 3 ml of trifluoroacetic acid. The mixture was stirred at room temperature under nitrogen for 2 hours. Solvent was removed and the residue was precipitated with ethyl acetate / hexanes to give 830 mg of the title compound (100%).
MS (FD) m / z 277 (M &lt; + &gt; for free amines).
IR (KBr) 3145, 1778, 1665, 1646, 1626 cm ^-1 .
<Example 7>
1- (3-aminopropyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one trifluoroacetate
1- (Nt-butyloxycarbonyl-3-aminopropyl) -isoxazolo [3,4-c] -1,2-di by the method of Example 6 except that the reaction time was 3 hours 922 mg (2.35 mmol) of hydro-3-methyl-6-chloroquinolin-2-one were converted to 837 mg of the title compound (88%).
Elemental Analysis for C ₁₆ H ₁₅ ClF ₃ N ₃ O ₄ (EA):
Calc .: C, 47.36; H, 3.73; N, 10.36.
Found: C, 47.44; H, 3. 70; N, 10.34.
MS (IS) m / z 292 (M &lt; + &gt; for free amines).
<Example 8>
1- (4-aminobutyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one trifluoroacetate
1- (Nt-butyloxycarbonyl-4-aminobutyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinoline- by the method of Example 7 3.14 g (7.75 mmol) of 2-one were converted to 2.90 g of the title compound (89%).
Elemental Analysis for C ₁₇ H ₁₇ ClF ₃ N ₃ O ₄ (EA):
Calc .: C, 48.64; H, 4.08; N, 10.01.
Found: C, 48.49; H, 3.95; N, 9.91.
MS (IS) m / z 306 (M &lt; + &gt; for free amines).
<Example 9>
1- (5-aminopentyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one trifluoroacetate
1- (Nt-butyloxycarbonyl-5-aminopentyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinoline- by the method of Example 7 1.08 g (2.57 mmol) of 2-one were converted to 1.01 g of the title compound (56%).
Elemental Analysis for C ₁₈ H ₁₉ ClF ₃ N ₃ O ₄ (EA):
Calc .: C, 49.84; H, 4.41; N, 9.69.
Found: C, 50.08; H, 4. 23; N, 9.69.
MS (IS) m / z 320 (M + for free amines).
<Example 10>
1- (6-aminohexyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one trifluoroacetate
1- (Nt-butyloxycarbonyl-6-aminohexyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinoline- by the method of Example 7 1.40 g (3.23 mmol) of 2-one were converted to 1.30 g of the title compound (90%).
Elemental Analysis for C ₁₉ H ₂₁ ClF ₃ N ₃ O ₄ (EA):
Calc .: C, 50.96; H, 4.73; N, 9.38.
Found: C, 51.20; H, 4. 62; N, 9.31.
MS (IS) m / z 334 (M &lt; + &gt; for free amines).
<Examples 11 and 12>
1- (N-trifluoroacetate-2-aminoethyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one and 1- ( N-3,4,5-trimethoxybenzoyl-2-aminoethyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
50 mg (0.128 mmol) of 1- (2-aminoethyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one trifluoroacetate Suspended in 3 ml anhydrous dichloromethane and 12.9 mg (0.128 mmol) N-methylmorpholine, and 1 ml anhydrous dimethylformamide was added. The resulting solution was stirred under nitrogen at room temperature, 59 mg (0.255 mmol) of 3,4,5-trimethoxybenzoyl chloride were added and the resulting mixture was stirred for about 18 hours. The reaction was worked up by adding ethyl acetate and water. Ethyl acetate was separated and washed three times with 1 N aqueous hydrochloric acid solution, sodium bicarbonate and 25 ml brine, respectively. Ethyl acetate was dried over sodium sulfate and filtered. Ethyl acetate was removed and the residue was chromatographed (silica gel, 1: 1 ethyl acetate: hexanes) to give 21 mg of trifluoroacetate amide title compound and 12 mg of benzoyl amide title compound (20%).
Trifluoroacetate amide: MS (IS) m / z 372 (M-H).
Benzoyl amide: MS (IS) m / z 472 (M &lt; + &gt;).
<Examples 13 and 14>
1- (N-trifluoroacetate-3-aminopropyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one and 1- ( N-3,4,5-trimethoxybenzoyl-3-aminopropyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
1- (3-aminopropyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one trifluoroacetate and 3,4,5- Trimethoxybenzoyl chloride (0.9 equiv) was converted to the title compound by the methods of Examples 11 and 12 to give 6 mg of trifluoroacetate amide title compound and 48 mg of benzoyl amide title compound (65%).
Trifluoroacetate amide: MS (IS) m / z 388 (M &lt; + &gt;).
Benzoyl amide: MS (FD) m / z 485 (M &lt; + &gt;). IR (KBr) 3284, 1673, 1634, 1585, 1339 cm ^-1 .
<Example 15>
1- (N-3,4,5-trimethoxybenzoyl-4-aminobutyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinoline-2 -On
75 mg (0.179 mmol) of 1- (4-aminobutyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one trifluoroacetate It was dissolved in 2 ml of anhydrous dimethylformamide and stirred under nitrogen at room temperature. 54 mg (0.537 mmol) of triethylamine were added and the solution turned purple. Then 62 mg (0.268 mmol) of 3,4,5-trimethoxybenzoyl chloride were added and the resulting mixture was stirred for about 18 hours. The reaction was diluted with ethyl acetate and washed three times each with 1N aqueous hydrochloric acid solution, sodium bicarbonate and brine 25 ml, dried over sodium sulfate, filtered and concentrated. The residue was chromatographed (silica gel, ethyl acetate) and the appropriate fractions recrystallized from dichloromethane to give 40 mg of the title compound (45%).
MS (IS) m / z 500 (M &lt; + &gt;).
IR (KBr) 1674, 1634, 1597, 1585, 1501 cm ^-1 .
<Examples 16 and 17>
1- (N-trifluoroacetate-5-aminopentyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one and 1- ( N-3,4,5-trimethoxybenzoyl-5-aminopentyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
1- (5-aminopentyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one trifluoroacetate and 3,4,5- Trimethoxybenzoyl chloride (0.9 equiv) was converted to the title compound by the methods of Examples 11 and 12 to yield 0.7 mg of trifluoroacetate amide title compound and 41 mg of benzoyl amide title compound (51%).
Trifluoroacetate amide: MS (IS) m / z 416 (M &lt; + &gt;).
Benzoyl amide: MS (FD) m / z 513 (M &lt; + &gt;).
IR (KBr) 1667, 1629, 1597, 1580, 1500 cm ^-1 .
<Examples 18 and 19>
1- (N-trifluoroacetate-6-aminohexyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one and 1- ( N-3,4,5-trimethoxybenzoyl-6-aminohexyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
1- (6-aminohexyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one trifluoroacetate and 3,4,5- Trimethoxybenzoyl chloride (0.9 equiv) was converted to the title compound by the methods of Examples 11 and 12 to give 10.7 mg of trifluoroacetate amide title compound and 36 mg of benzoyl amide title compound (45%).
Trifluoroacetate Amide:
Elemental Analysis for C ₁₉ H ₁₉ ClF ₃ N ₃ O ₃ (EA):
Calc .: C, 53.09; H, 4. 46; N, 9.78.
Found: C, 53.34; H, 4. 37; N, 9.72.
MS (IS) m / z 430 (M &lt; + &gt;).
Benzoyl amide:
Elemental Analysis for C ₂₇ H ₃₀ ClF ₃ N ₃ O ₆ (EA):
Calc .: C, 61.42; H, 5.73; N, 7.96.
Found: C, 61.25; H, 5.79; N, 7.69.
MS (FD) m / z 528 (M &lt; + &gt;).
<Example 20>
1- (3-carboxypropyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
Methyl-N- (3- [2-chloro-6-fluorophenyl] -5-methylisoxazol-4-oyl) -4 by the method of Example 1 except that the reaction time was 3 hours 1.54 g (4.35 mmol) of aminobutanoate were converted to 1.15 g of the title compound (82%).
MS (FD) m / z 320 (M &lt; + &gt;).
IR (KBr) 3145, 2925, 1740, 1732, 1651, 1627, 1598 cm ^-1 .
<Example 21>
1- (4-carboxybutyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
By the method of Example 20 200 mg (0.542 mmol) of methyl-N- (3- [2-chloro-6-fluorophenyl] -5-methylisoxazol-4-oil) -5-aminopentanoate were added. The title compound was converted to 180 mg (100%).
Elemental Analysis for C ₁₆ H ₁₅ ClN ₂ O ₄ (EA):
Calc .: C, 57.41; H, 4.52; N, 8.37.
Found: C, 57.43; H, 4.53; N, 8.08.
MS (FD) m / z 334.1 (M &lt; + &gt;).
<Example 22>
1- (5-carboxypentyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
By the method of Example 21, 735 mg (1.92 mmol) of methyl-N- (3- [2-chloro-6-fluorophenyl] -5-methylisoxazol-4-oyl) -6-aminohexanoate Was converted to 499 mg of the title compound (74%).
Elemental Analysis for C ₁₇ H ₁₇ ClN ₂ O ₄ (EA):
Calc .: C, 58.54; H, 4.91; N, 8.03.
Found: C, 58.51; H, 4.99; N, 7.89.
MS (FD) m / z 348 (M &lt; + &gt;).
<Example 23>
1- (6-carboxyhexyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one
By the method of Example 21, 1.49 g (3.75 mmol) of methyl-N- (3- [2-chloro-6-fluorophenyl] -5-methylisoxazol-4-oyl) -7-aminoheptanoate Was converted to 1.07 g of the title compound (79%).
Elemental Analysis for C ₁₈ H ₁₉ ClN ₂ O ₄ (EA):
Calc .: C, 59.59; H, 5. 28; N, 7.72.
Found: C, 59.66; H, 5.49; N, 7.64.
MS (FD) m / z 362 (M &lt; + &gt;).
<Example 24>
1- (4-oxo-4- (3,4,5-trimethoxyphenylamino) butyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloro Quinolin-2-one
By the method of Example 26, 5- (isooxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one-1-yl) butanoic acid and 3 , 4,5-trimethoxyaniline was converted to the title compound to give 67 mg (88%).
Elemental Analysis for C ₂₄ H ₂₄ ClN ₃ O ₆ (EA):
Calc .: C, 59.32; H, 4.98; N, 5.12.
Found: C, 59.52; H, 5. 12; N, 8.41.
MS (FD) m / z 484.9 (M−H).
<Example 25>
1- (4-oxo-4- (3,4,5-trimethoxybenzylamino) butyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloro Quinolin-2-one
By the method of Example 26, 5- (isooxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one-1-yl) butanoic acid and 3 , 4,5-trimethoxybenzylaniline was converted to the title compound to give 37 mg (47%).
Elemental Analysis for C ₂₄ H ₂₄ ClN ₃ O ₆ (EA):
Calc .: C, 60.06; H, 5.24.
Found: C, 60.20; H, 5.49.
MS (FD) m / z 498.9 (M−H).
<Example 26>
1- (5-oxo-5- (3,4,5-trimethoxyphenylamino) pentyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloro Quinolin-2-one
46 mg (0.137 mmol) of 1- (4-carboxybutyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one were dissolved in anhydrous dichloromethane 3 Dissolve in mL and stir under nitrogen at room temperature. 26 mg (0.137 mmol) of 1-ethyl-3- [3- (dimethylamino) propyl] carbodiimide hydrochloride (EDCI) were added followed by 25 mg (0.137 mmol) of 3,4,5-trimethoxyaniline. Was added and the resulting mixture was stirred for about 18 hours. TLC (ethyl acetate) indicates that the reaction was substantially incomplete, thus adding a catalytic amount of dimethylaminopyridine and stirring the reaction for about 18 hours. The reaction was diluted with ethyl acetate, washed three times each with 1N aqueous hydrochloric acid solution, sodium bicarbonate, brine, dried over sodium sulfate, filtered and ethyl acetate removed to give 41 mg of the title compound (60%).
Elemental Analysis for C ₂₅ H ₂₆ ClN ₃ O ₆ (EA):
Calc .: C, 60.06; H, 5. 24; N, 8.41.
Found: C, 59.94; H, 5.47; N, 8.14.
MS (FD) m / z 498.9 (M−H).
<Example 27>
1- (5-oxo-5- (3,4,5-trimethoxybenzylamino) pentyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloro Quinolin-2-one
40 mg (0.119 mmol) of 1- (4-carboxybutyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one were dissolved in anhydrous dichloromethane 3 Dissolve in mL and stir under nitrogen at room temperature. 23 mg (0.119 mmol) of EDCI were added to make the reaction a solution. 23.6 mg (0.119 mmol) of 3,4,5-trimethoxybenzylamine and catalytic amount of DMAP were added and the reaction mixture was stirred for about 18 hours. TLC (ethyl acetate) showed that the reaction was substantially complete. The reaction was diluted with ethyl acetate, washed three times each with 1N aqueous hydrochloric acid solution, sodium bicarbonate, brine, dried over sodium sulfate, filtered and ethyl acetate removed to give 43 mg (70%) of the title compound.
Elemental Analysis for C ₂₆ H ₂₈ ClN ₃ O ₆ (EA):
Calc .: C, 60.76; H, 5.49; N, 8.18.
Found: C, 60.89; H, 5. 66; N, 7.93.
MS (FD) mlz 512.8 (M-H).
<Example 28>
1- (6-oxo-6- (3,4,5-trimethoxyphenylamino) hexyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloro Quinolin-2-one
5- (isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one-1-yl) hexanoic acid and 3,4,5-trimethoxy Aniline was converted to the title compound by the method of Example 26 to give 56 mg (77%).
Elemental Analysis for C ₂₆ H ₂₈ ClN ₃ O ₆ (EA):
Calc .: C, 60.76; H, 5.49; N, 8.17.
Found: C, 60.18; H, 5. 34; N, 7.82.
MS (FD) m / z 513.0 (M-H).
<Example 29>
1- (6-oxo-6- (3,4,5-trimethoxybenzylamino) hexyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloro Quinolin-2-one
5- (isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one-1-yl) hexanoic acid and 3,4,5-trimethoxy Aniline was converted to the title compound by the method of Example 26 to give 63 mg (83%).
Elemental Analysis for C ₂₇ H ₃₀ ClN ₃ O ₆ (EA):
Calc .: C, 61.42; H, 5.73; N, 7.96.
Found: C, 61.32; H, 5.67; N, 7.68.
MS (FD) m / z 527.0 (M−H).
<Example 30>
1- (7-oxo-7- (3,4,5-trimethoxyphenylamino) heptyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloro Quinolin-2-one
5- (isooxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one-1-yl) heptanoic acid and 3,4,5-trimethoxy Aniline was converted to the title compound by the method of Example 26 to give 52 mg (71%).
Elemental Analysis for C ₂₇ H ₃₀ ClN ₃ O ₆ (EA):
Calc .: C, 61.42; H, 5.73; N, 7.96.
Found: C, 61.34; H, 5. 82; N, 7.68.
MS (FD) m / z 526.9 (M−H).
<Example 31>
1- (7-oxo-7- (3,4,5-trimethoxybenzylamino) heptyl) -isoxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloro Quinolin-2-one
5- (isooxazolo [3,4-c] -1,2-dihydro-3-methyl-6-chloroquinolin-2-one-1-yl) heptanoic acid and 3,4,5-trimethoxy Benzylamine was converted to the title compound by the method of Example 26 to give 69 mg (92%).
Elemental Analysis for C ₂₈ H ₃₂ ClN ₃ O ₆ (EA):
Calc .: C, 62.05; H, 5.95; N, 7.75.
Found: C, 61.78; H, 5. 82; N, 7.49.
MS (FD) m / z 541 (M−H).
Compounds of the invention are inhibitors of MRP1. Thus, the compounds of the present invention can be used to inhibit any tumors with innate and / or acquired resistance to tumor cell disruptor (s) conferred in part or wholly by MRP1. In other words, treatment of the tumor with an effective amount of a compound of the present invention makes the tumor more sensitive to chemotherapy that is less potent by MRP1.
Vincristine, epirubicin, daunorubicin, doxorubicin and etoposide are tumor cell disruptors that are substrates of MRP1 [see, eg, Cole, et.al., "Pharmacological Characterization of Multidrug Resistant MRP-transfected Human Tumor Cells". , Cancer Research, 54: 5902-5910, 1994]. Since MRP1 is ubiquitous in mammals, especially humans, see Nuter, K, et. al., "Expression of the Multidrug Resistance-Associated Protein (MRP) Gene in Human Cancers", Clin. Can. Res., 1: 1301-1310 (1995)], where chemotherapy whose purpose is to inhibit tumors using any of these agents has the potential to be less effective by MRP1. Thus, tumors of the bladder, bone, breast, lung (small cell), testes and thyroid and more specific types of cancers, such as acute lymphoblastic and myeloid leukemia, Wilms' tumor, neuroblastoma, soft tissue sarcoma, Hodgkin's And non-Hodgkin's lymphomas and organ supportive cancers can be inhibited by the combination of one or more of the above tumor cell disruptors and the compounds of the present invention.
The biological activity of the compounds of the invention was assessed using an initial screening assay that measured quickly and accurately the activity of the compounds tested in inhibiting MRP1 or MDR1. Assays useful for assessing this reversal ability are known in the art [eg, T. McGrath, et al., Biochemical Pharmacology, 38: 3611, 1989; D. Marquardt and M.S. Center, Cancer Research, 52: 3157, 1992; D. Marquardt, et al., Cancer Research, 50: 1426, 1990; And Cole, et.al., "Pharmacological Characterization of Multidrug Resistant MRP-transfected Human Tumor Cells", Cancer Research, 54: 5902-5910, 1994].
Assays for reversal of MRP1-mediated doxorubicin resistance and MDR1-mediated vincristine resistance:
HL60 / ADR and HL60 / VCR are continuous cell lines and doxorubicin and vincristine resistance by culturing human acute myeloid leukemia cell line HL60, respectively, in increasing concentrations of doxorubicin or vincristine until a highly resistant mutant is obtained. Were screened for.
HL60 / ADR and HL60 / VCR cells were grown in RPMI 1640 (Gibco) containing 10% fetal bovine serum (FBS) and 250 μg / ml GENTAMICIN ^™ (Sigma). Cells are harvested and washed twice with assay medium (same as medium); Counting; Dilute to 2 x 10 ⁵ cells / ml in assay medium. 50 microliters of cells were aliquoted into the wells of a 96 well tissue culture plate. One column of each 96 well plate was used as a negative control and medium containing no cells was added.
Test compounds and reference compounds were dissolved in dimethyl sulfoxide (DMSO) at a concentration of 5 mM. Samples were diluted to 20 μM in assay medium and 25 μl of each test compound was added to 6 wells. Assay standards were run in four replicates. 0.4 microliters of 25% DMSO was added to four wells as a solvent control. Assay medium was added to all wells so that the final volume per well was 100 μl.
Plates were incubated at 37 ° C. for 72 hours in a humidified incubator with 5% carbon dioxide atmosphere. Cell growth and viability were measured by oxidizing the tetrazolium salt using standard conditions. Plates were incubated at 37 ° C. for 3 hours. Absorbance was measured at 490 nm using a microtiter plate reader.
The test compound's ability to reverse the resistance of HL60 / ADR and HL60 / VCR cells to doxorubicin can be absorbed by wells containing tumor cell disintegrators without test compounds and by those containing test compounds in addition to tumor cell disintegrators. Was determined by comparison. A control was used to remove the background so that the results were not reliably artificial. The results of the assay are expressed as inhibition of cell growth. Tumor cell disintegrator alone at the concentrations tested generally did not inhibit the growth of HL60 / ADR or HL60 / VCR cells.
Representative compounds of formula (I) have demonstrated significant effects in reversing MRP1 multidrug resistance. Many compounds have shown very significant improvement in activity with tumor cell disruptors as opposed to tumor cell disruptors alone. In addition, the majority of the compounds tested showed a significant degree of selective inhibition of the HL60 / ADR cell line rather than the HL60 / VCR cell line.
In practicing the methods of the present invention, when administering a tumor cell disruptor, the amount of tumor cell disruptor used is variable. The amount of tumor cell disruptor actually administered includes the disease to be treated, the route of administration chosen, the actual tumor cell disruptor administered, the age, weight and response of the individual patient (mammal), and the depth of symptoms of the patient. It should be understood that the decision may be made by a physician in light of the circumstances involved. Of course, the amount of tumor cell disruptor administered should be determined by the patient's physician and closely monitored. After deciding which tumor cell disintegrator to use, "The Physician's Desk Reference", published by Medical Economics Company at Montvale, NJ 07645-1742, determines the amount of tumor cell disintegrator to be administered. This can help doctors, and the literature is updated annually.
Preferred agents and methods of the invention using these agents are those that do not contain a tumor cell disruptor. Therefore, it is preferable to administer the compound of the present invention separately from the tumor cell disruptor. Tumor cell disintegrators referred to herein are commercially available and may be purchased in preformulation form suitable for the methods of the present invention.
Compounds of formula (I) alone or optionally in admixture with tumor cell disruptors are generally administered in the form of pharmaceutical preparations. These agents can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal. Such formulations are prepared in a manner known in the pharmaceutical art and comprise one or more active compounds of formula (I).
The invention also includes a method of using a pharmaceutical formulation containing a pharmaceutical carrier with the compound of formula (I) and optionally a tumor cell disrupting agent as the active ingredient. In preparing the formulations of the present invention, the active ingredient (s) are generally included in a carrier, which may be mixed with an excipient, diluted with an excipient, or in the form of a capsule, small pouch, paper or other container. When an excipient is used as a diluent, it may be a solid, semisolid or liquid substance which acts as a carrier, carrier or medium for the active ingredient. Thus, the formulations may contain tablets, pills, powders, sugar-containing tablets, small bags, cachets, elixir suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), for example up to 10% by weight of active compound. Ointments, soft and hard gelatin capsules containing, suppositories, sterile injectable solutions, and sterile packaged powders.
In preparing the formulations, it is necessary to mill the active compound (s) to provide the appropriate particle size before combining with the other components. If the active compound (s) are substantially insoluble, they are usually milled to a particle size of less than 200 mesh. If the active compound (s) are substantially water soluble, the particle size is usually adjusted, for example by milling to about 40 mesh, to provide a substantially uniform distribution in the formulation.
Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, Water, syrup, and methyl cellulose. The formulations may additionally include lubricants such as talc, magnesium stearate, and mineral oils; Wetting agents; Emulsifying and suspending agents; Preservatives such as methyl- and propylhydroxybenzoate; Sweeteners; And flavoring agents. The formulations of the present invention may be formulated to provide rapid release, sustained release or sustained release of the active ingredient after administration to a patient using methods known in the art.
The formulations are preferably formulated in unit dosage forms in which each dosage unit contains about 5 to about 100 mg, more generally about 10 to about 30 mg of each active ingredient. The term "unit dosage form" means that the physically separated into a single dosage form suitable for humans and other mammals, each unit containing a predetermined amount of active substance calculated to produce the desired therapeutic effect, with a suitable pharmaceutical excipient Say unit.
Compounds of formula (I) are effective over a wide dosage range. For example, the dosage per day is usually in the range of about 0.5 to about 30 mg per kg of body weight. In the treatment of adults, about 1 to about 15 mg / kg / day in single or divided doses is particularly preferred. However, the amount of compound actually administered may be determined by the physician in light of the relevant circumstances including the disease to be treated, the route of administration chosen, the actual compound administered, the age, weight and response of the individual patient, and the depth of the patient's symptoms and Therefore, it should be understood that the above dosage ranges are not intended to limit the scope of the invention in any sense. In some cases, dosages below the lower limit of the above range may be more appropriate, while in other cases, higher dosages may be any harmful as long as more dosages are divided into several smaller dosages for administration over the day. Can be used without side effects.
To prepare solid preparations, such as tablets, the main active ingredient (s) are mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogeneous mixture of the compounds of the invention. When referring to these preformulation compositions as homogeneous, this allows the active ingredient (s) to be uniformly dispersed throughout the formulation so that the formulation can be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. The solid preform is then subdivided into unit dosage forms of the type described above containing from 0.1 to about 500 mg of the active ingredient of the invention.
The tablets or pills of the present invention may be coated or otherwise formulated to provide dosage forms that provide the benefit of sustaining action. For example, tablets or pills may include internal and external dosage components, where the external dosage component is in the form of an envelope above the internal dosage component. The two components can be separated by an enteric layer that withstands disintegration in the stomach, allowing the internal components to pass through the duodenum or be released free of charge. Various materials can be used in such enteric layers or coatings, and such materials include many polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol and cellulose acetate.
New formulations, which may be in liquid form, may be incorporated to be administered orally or by injection, and may include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and edible oils such as cottonseed oil, sesame oil, coconut oil, Or flavored emulsions with peanut oil, as well as elixirs and similar pharmaceutical excipients.
Preparations for inhalation or insufflation include solutions and suspensions in pharmaceutical aqueous or organic solvents, or mixtures and powders thereof. Liquid or solid preparations may contain suitable pharmaceutical excipients as described above. Preferably, the formulation is administered by the oral or nasal respiratory route for local or systemic effects. Preferably the composition in the pharmaceutical solvent can be nebulized by the use of an inert gas. The nebulization solution may breathe directly from the nebulization apparatus or attach the nebulization apparatus to a face mask, tent or intermittent amount of pressure respirator. The solution, suspension or powder formulation may be administered orally or intranasally, preferably from a device that delivers the formulation in a suitable manner.
The following formulation examples are illustrative only and are not intended to limit the scope of the invention in any sense. "Active ingredient (s)" means a compound according to formula (I) or a pharmaceutical salt or solvate thereof and optionally at least one tumor cell disruptor.
<Example 1>
Hard gelatin capsules were prepared containing the following ingredients:
ingredientVolume (mg / capsule) Active ingredient (s)30.0 Starch305.0 Magnesium stearate5.0
The ingredients were mixed and filled into hard gelatin capsules in an amount of 340 mg.
<Example 2>
Tablet formulations were prepared using the following ingredients:
ingredientVolume (mg / tablet) Active ingredient (s)25.0 Cellulose, microcrystalline200.0 Colloidal silicon dioxide10.0 Stearic acid5.0
The ingredients were blended and compressed to produce tablets weighing 240 mg each.
<Example 3>
A dry powder inhaler formulation was prepared containing the following ingredients:
ingredientweight % Active ingredient (s)5 Lactose95
The active ingredient is mixed with lactose and the mixture is added to a dry powder inhaler.
<Example 4>
Tablets each containing 30 mg of the active ingredient were prepared as follows:
ingredientVolume (mg / tablet) Active ingredient (s)30.0 mg Starch45.0 mg Microcrystalline cellulose35.0 mg Polyvinylpyrrolidone (with 10% solution in water)4.0 mg Sodium carboxymethyl starch4.5 mg Magnesium stearate0.5 mg talc1.0 mg Sum120 mg
The active ingredient, starch and cellulose are no. 20 mesh U.S. Pass the sieve and mix thoroughly. A solution of polyvinylpyrrolidone was mixed with the resulting powder and then 16 mesh U.S. Passed through a sieve. The granules thus prepared are dried at 50-60 ° C. and 16 mesh U.S. Passed through a sieve. No. in advance 30 mesh U.S. Sodium carboxymethyl starch, magnesium stearate, and talc, which were passed through a sieve, were added to the granules, which were mixed on a tableting machine after mixing to obtain tablets each weighing 120 mg.
<Example 5>
40 mg of each medicament was prepared as follows:
ingredientVolume (mg / capsule) Active ingredient (s)40.0 mg Starch109.0 mg Magnesium stearate1.0 mg Sum150.0 mg
The active ingredient, cellulose, starch and magnesium stearate are blended and No. 20 mesh U.S. The sieve was passed through and filled into hard gelatin capsules in an amount of 150 mg.
<Example 6>
Suppositories containing 25 mg of each active ingredient were prepared as follows:
ingredientamount Active ingredient (s)25 mg Saturated Fatty Acid GlycerideAmount to total 2,000 mg
Active ingredient No. 6 mesh U.S. The sieve was passed through and suspended in saturated fatty acid glycerides pre-melted using the minimum heat required. The mixture was then poured into a suppository mold of nominal 2.0 g capacity and allowed to cool.
<Example 7>
Suspensions containing 50 mg of drug per 5.0 ml of each dose were prepared as follows:
ingredientamount Active ingredient (s)50.0 mg Xanthan gum4.0 mg Sodium Carboxymethyl Cellulose (11%) Microcrystalline Cellulose (89%)50.0 mg Sucrose1.75 g Sodium benzoate10.0 mg Flavoring and Coloring AgentsQuantity Purified waterAmount to be 5.0 ml
The active ingredient, sucrose and xanthan gum are blended and No. 10 mesh U.S. The sieve was passed through and then mixed with a solution of sodium carboxymethyl cellulose and microcrystalline cellulose in previously prepared water. Sodium benzoate, flavors and colorants were diluted with some water and added with stirring. Sufficient water was then added to make the required volume.
<Example 8>
Capsules containing 15 mg of each drug were prepared as follows:
ingredientVolume (mg / tablet) Active ingredient (s)15.0 mg Starch407.0 mg Magnesium stearate3.0 mg Sum425.0 mg
The active ingredient, cellulose, starch and magnesium stearate are blended and No. 20 mesh U.S. The sieve was passed through and filled into hard gelatin capsules in an amount of 425 mg.
<Example 9>
Intravenous preparations can be prepared as follows:
ingredientVolume (mg / tablet) Active ingredient (s)250.0 mg Isotonic saline1000 ml
<Example 10>
Topical formulations can be prepared as follows:
ingredientVolume (mg / tablet) Active ingredient (s)1-10 g Oil painting wax30 g Liquid paraffin20 g White soft paraffinThe amount that adds up to 100 g
The white soft paraffin was heated until it melted. Liquid paraffin and emulsified wax were incorporated and stirred until dissolved. Stirring was continued until the active ingredient was added and dispersed. The mixture was then cooled until it became a solid.
<Example 11>
Sublingual or buccal tablets each containing 10 mg of the active ingredient may be prepared as follows:
ingredientAmount per tablet Active ingredient (s)10.0 mg Glycerol210.5 mg water143.0 mg Sodium citrate4.5 mg Polyvinyl alcohol26.5 mg Polyvinylpyrrolidone15.5 mg Sum410.0 mg
Glycerol, water, sodium citrate, polyvinyl alcohol and polyvinylpyrrolidone were mixed together with continuous stirring and maintaining the temperature at about 90 ° C. When the polymer was in solution, the solution was cooled to about 50-55 ° C. and the active ingredients were slowly mixed. The homogeneous mixture was poured into moldings made of inert material to prepare a drug containing dispersion matrix having a thickness of about 2-4 mm. This dispersion matrix was then cut to form individual tablets of the appropriate size.
Other preferred agents used in the methods of the present invention use transdermal delivery devices ("patches"). Such transdermal patches can be used to continuously or discontinuously inject the compounds of the present invention in controlled amounts. The construction and use of transdermal patches for the delivery of pharmaceutical agents are known in the art. See, for example, US Pat. No. 5,023,252, issued June 11, 1991, which is incorporated herein by reference. ]. Such patches may be configured for continuous, pulsatile, or on demand delivery of pharmaceutical agents.
Often, it is desirable or necessary to introduce pharmaceutical agents directly or indirectly into the brain. Direct techniques generally include placing a drug delivery catheter in the ventricle of the host to bypass the blood transfusion gateway. One such implantable delivery system used to transport biological factors to specific anatomical regions of the body is described in US Pat. No. 5,011,472, issued April 30, 1991, which is incorporated herein by reference.
Generally preferred indirect techniques typically include formulating the composition to provide drug latentiation by converting the hydrophilic drug into a fat soluble drug or a prodrug. Latency is generally achieved through blocking of hydroxy, carbonyl, sulfate, and primary amine groups present on the drug to make the drug more soluble and transportable through the blood transfusion gateway. Alternatively, delivery of hydrophilic drugs may be enhanced by intraarterial injection of hypertonic solution that may temporarily open the blood-brain barrier.

权利要求:
Claims (13)
[1" claim-type="Currently amended] A compound of formula (I) or a pharmaceutical salt or solvate thereof.
<Formula I>

Where
m is an integer from 1 to 6,
R is COR ¹ , amino, NH-Pg or NHCOR ² ,
R ¹ is hydroxy, C ₁ -C ₆ alkoxy, or NR ³ R ⁴ ,
Pg is an amino protecting group,
R ² is C ₁ -C ₆ alkyl, substituted C ₁ -C ₄ alkyl, aryl, substituted aryl, (CH ₂ ) _n -heterocycle, (CH ₂ ) _n -substituted heterocycle,
R ³ is independently at each occurrence hydrogen or C ₁ -C ₆ alkyl,
R ⁴ is C ₁ -C ₆ alkyl, norbornan-2-yl, aryl, substituted aryl, CH ₂ CH (CH ₃ ) phenyl, (CH ₂ ) _n heterocycle or (CH ₂ ) _n -substituted hetero Cycle,
n is 0, 1 or 2.
[2" claim-type="Currently amended] The compound of claim 1, wherein m is an integer of 2 to 6, R is t-butyloxycarbonylamino, trifluoroacetylamino, 3,4,5-trimethoxybenzoylamino, 3,4,5- A compound selected from trimethoxyanilinylcarboxy and 3,4,5-trimethoxybenzylaminilcarboxy or a pharmaceutical salt or solvate thereof.
[3" claim-type="Currently amended] A method of inhibiting MRP1 in a mammal comprising administering an effective amount of a compound of Formula (I) or a pharmaceutical salt or solvate thereof to a mammal in need of inhibition of MRP1.
<Formula I>

Where
m is an integer from 1 to 6,
R is COR ¹ , amino, NH-Pg or NHCOR ² ,
R ¹ is hydroxy, C ₁ -C ₆ alkoxy, or NR ³ R ⁴ ,
Pg is an amino protecting group,
R ² is C ₁ -C ₆ alkyl, substituted C ₁ -C ₄ alkyl, aryl, substituted aryl, (CH ₂ ) _n -heterocycle, (CH ₂ ) _n -substituted heterocycle,
R ³ is independently at each occurrence hydrogen or C ₁ -C ₆ alkyl,
R ⁴ is C ₁ -C ₆ alkyl, norbornan-2-yl, aryl, substituted aryl, CH ₂ CH (CH ₃ ) phenyl, (CH ₂ ) _n heterocycle or (CH ₂ ) _n -substituted hetero Cycle,
n is 0, 1 or 2.
[4" claim-type="Currently amended] The method of claim 3, wherein the mammal is a human.
[5" claim-type="Currently amended] The compound of formula I, wherein m is an integer from 2 to 6, R is t-butyloxycarbonylamino, trifluoroacetylamino, 3,4,5-trimethoxybenzoylamino, 3 And 4,5-trimethoxyanilinylcarboxy and 3,4,5-trimethoxybenzylaminylcarboxy or a pharmaceutical salt or solvate thereof.
[6" claim-type="Currently amended] Administering an effective amount of a compound of formula (I) or a pharmaceutical salt or solvate thereof in combination with an effective amount of one or more tumor cell disintegrants to a mammal in need thereof: A method of inhibiting a resistant tumor or a tumor prone to resistance in a mammal.
<Formula I>

Where
m is an integer from 1 to 6,
R is COR ¹ , amino, NH-Pg or NHCOR ² ,
R ¹ is hydroxy, C ₁ -C ₆ alkoxy, or NR ³ R ⁴ ,
Pg is an amino protecting group,
R ² is C ₁ -C ₆ alkyl, substituted C ₁ -C ₄ alkyl, aryl, substituted aryl, (CH ₂ ) _n -heterocycle, (CH ₂ ) _n -substituted heterocycle,
R ³ is independently at each occurrence hydrogen or C ₁ -C ₆ alkyl,
R ⁴ is C ₁ -C ₆ alkyl, norbornan-2-yl, aryl, substituted aryl, CH ₂ CH (CH ₃ ) phenyl, (CH ₂ ) _n heterocycle or (CH ₂ ) _n -substituted hetero Cycle,
n is 0, 1 or 2.
[7" claim-type="Currently amended] The method of claim 6, wherein the mammal is a human.
[8" claim-type="Currently amended] 8. The compound of formula I according to claim 7, wherein m is an integer from 2 to 6, R is t-butyloxycarbonylamino, trifluoroacetylamino, 3,4,5-trimethoxybenzoylamino, 3 And 4,5-trimethoxyanilinylcarboxy and 3,4,5-trimethoxybenzylaminylcarboxy or a pharmaceutical salt or solvate thereof.
[9" claim-type="Currently amended] 8. The method of claim 7, wherein the tumor cell disintegrant is selected from doxorubicin, daunorubicin, epirubicin, vincristine and etoposide.
[10" claim-type="Currently amended] 8. The method of claim 7, wherein the tumor is of Wilms type, bladder, bone, breast, lung (small cell), testis or thyroid, or the tumor is acute lymphoblastic and myeloid leukemia, neuroblastoma, soft tissue sarcoma, Method associated with Hodgkin's and non-Hodgkin's lymphoma or organ support cancer.
[11" claim-type="Currently amended] A pharmaceutical formulation comprising a compound of formula (I) or a pharmaceutical salt or solvate thereof in combination with one or more pharmaceutical carriers, diluents or excipients.
<Formula I>

Where
m is an integer from 1 to 6,
R is COR ¹ , amino, NH-Pg or NHCOR ² ,
R ¹ is hydroxy, C ₁ -C ₆ alkoxy, or NR ³ R ⁴ ,
Pg is an amino protecting group,
R ² is C ₁ -C ₆ alkyl, substituted C ₁ -C ₄ alkyl, aryl, substituted aryl, (CH ₂ ) _n -heterocycle, (CH ₂ ) _n -substituted heterocycle,
R ³ is independently at each occurrence hydrogen or C ₁ -C ₆ alkyl,
R ⁴ is C ₁ -C ₆ alkyl, norbornan-2-yl, aryl, substituted aryl, CH ₂ CH (CH ₃ ) phenyl, (CH ₂ ) _n heterocycle or (CH ₂ ) _n -substituted hetero Cycle,
n is 0, 1 or 2.
[12" claim-type="Currently amended] (a) a compound of formula (I) or a pharmaceutical salt or solvate thereof,
(b) at least one tumor cell disruptor, and
(c) one or more pharmaceutical carriers, diluents or excipients
Pharmaceutical formulations comprising a.
<Formula I>

Where
m is an integer from 1 to 6,
R is COR ¹ , amino, NH-Pg or NHCOR ² ,
R ¹ is hydroxy, C ₁ -C ₆ alkoxy, or NR ³ R ⁴ ,
Pg is an amino protecting group,
R ² is C ₁ -C ₆ alkyl, substituted C ₁ -C ₄ alkyl, aryl, substituted aryl, (CH ₂ ) _n -heterocycle, (CH ₂ ) _n -substituted heterocycle,
R ³ is independently at each occurrence hydrogen or C ₁ -C ₆ alkyl,
R ⁴ is C ₁ -C ₆ alkyl, norbornan-2-yl, aryl, substituted aryl, CH ₂ CH (CH ₃ ) phenyl, (CH ₂ ) _n heterocycle or (CH ₂ ) _n -substituted hetero Cycle,
n is 0, 1 or 2.
[13" claim-type="Currently amended] 13. The formulation of claim 12, wherein the tumor cell disruptor is selected from doxorubicin, daunorubicin, epirubicin, vincristine and etoposide.

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同族专利:

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公开号 | 公开日

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NO20005042L|2000-10-06|

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EP1069897A1|2001-01-24|

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EA200001036A1|2001-04-23|

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CA2327706A1|1999-10-14|

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JP2002510624A|2002-04-09|

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WO1999051227A1|1999-10-14|

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IL138391D0|2001-10-31|

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US6417193B1|2002-07-09|

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HU0101736A3|2002-04-29|

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HU0101736A2|2001-11-28|

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TR200002803T2|2000-12-21|

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NO20005042D0|2000-10-06|

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SK15002000A3|2001-06-11|

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AU3468099A|1999-10-25|

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BR9909497A|2000-12-12|

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PL343312A1|2001-08-13|

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CN1296408A|2001-05-23|

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ID27048A|2001-02-22|

引用文献:

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公开号 | 申请日 | 公开日 | 申请人 | 专利标题

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法律状态:
1998-04-08|Priority to US8107798P

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1998-04-08|Priority to US60/081,077

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1999-04-07|Application filed by 피터 지. 스트링거, 일라이 릴리 앤드 캄파니

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1999-04-07|Priority to PCT/US1999/007343

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2001-05-25|Publication of KR20010042482A

优先权:

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申请号 | 申请日 | 专利标题

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US8107798P| true| 1998-04-08|1998-04-08|

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US60/081,077|1998-04-08|

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PCT/US1999/007343|WO1999051227A1|1998-04-08|1999-04-07|Methods for inhibiting mrp1|