Tyrosine Kinase Inhibitors

Abstract

The present disclosure is directed, in part, to a method of treating cancer, such as a B cell cancer, in a patient in need thereof, comprising administering to the patient an effective amount of a disclosed tyrosine kinase inhibitor. In some embodiments, the cancer harbors a BTK mutation.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of and priority to U.S. Application No. 63 / 328,348, filed April 7, 2022, the contents of which are incorporated herein by reference in their entirety. [Background technology]

[0002] Bruton's tyrosine kinase (BTK) inhibition is a novel strategy in the treatment of B-cell malignancies. Ibrutinib is the most studied BTK inhibitor and the first agent of this new class approved by the US Food and Drug Administration (FDA) and the European Medicines Agency (EMA). Resistance to ibrutinib treatment is due to the selection of cells harboring pathogenic mutations that alter BTK. The most common resistance mutation is a cysteine ​​(C) to serine (S) substitution at position 481, which prevents covalent binding of ibrutinib to the thiol group at the ATP binding site. When this mutation was introduced into the germline of mice, B-cell development remained normal, demonstrating functional compatibility. The BTK mutations C481F, C481G, C481R, and C481Y are highly prevalent in a subset of CLL patients, but are much less frequent than C481S.

[0003] Acalabrutinib is a second-generation BTK inhibitor that binds covalently to wild-type C481. It has greater selectivity and fewer side effects than ibrutinib. Acalabrutinib is FDA-approved for the treatment of mantle cell lymphoma (MCL) and CLL / small lymphocytic leukemia. Zanubrutinib is also a more selective and irreversible BTK inhibitor that is FDA-approved for the treatment of MCL. Zanubrutinib shows potent preclinical activity and minimal off-target effects in patients with Waldenström's macroglobulinemia.

[0004] Reversible non-covalent inhibitors are also selective for BTK and do not bind C481, so inhibitory activity is likely to be at least partially maintained in the presence of the C481S mutant. Non-covalent inhibitors have shown high potency against BTK mutants, including C481R and T474I / M, in in vitro assays. The non-covalent BTK inhibitor fenebrutinib has proven safe and demonstrated comparable BTK inhibition against wild-type and C481S mutants and is in phase I trials. Summary of the Invention [Problem to be solved by the invention]

[0005] Thus, there is a need for the development of BTK inhibitors and methods for the treatment of diseases or conditions that respond to BTK inhibition, such as B cell malignancies, to reduce side effects, and to overcome resistance to, for example, ibrutinib treatment. [Means for solving the problem]

[0006] The present disclosure is directed, at least in part, to a method of treating cancer with compounds that modulate, e.g., inhibit, Bruton's tyrosine kinase (BTK). For example, disclosed herein is a method of treating cancer in a patient in need thereof, comprising administering to the patient an effective amount of a Bruton's tyrosine kinase (BTK) inhibitor represented by formula (I) or a pharma- ceutically acceptable salt or stereoisomer thereof. TIFF2025514034000002.tif3282 formula, R 1 NHR a and N.R. a R b is selected from the group consisting of: R a and R b are each independently selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C3-C6 alkynyl, C3-C6 cycloalkyl, and C3-C6 cycloalkenyl, or R a and R bmay be taken together with the nitrogen to which they are attached to form a C3-C6 heterocyclyl. R 2 OR 11 , Hydrogen, Halo, NHR 11 , C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl. R 3 is NHCO2R 4 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, aryl, halo, aryloxy, NH(CO)NR 5 R 6 , NH(CO)R 7 , NH-C1-C6 alkyl, NH-C2-C6 alkenyl, NH(CH2) n -aryl, (CH2) p -heteroaryl, (CH2) q CO2R 8 , (CH2) r COR 9 and NHSO2R 10 wherein each C1-C6 alkyl, C2-C6 alkenyl, aryl or heteroaryl group in the preceding list is independently selected from the group consisting of C1-C6 alkyl, halo, OH, NR c R d ,CONR c R d , C1-C6 alkoxy, aryloxy, and CO2H. R 4 From R 11 are each independently selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, and aryl. R c and R d are each independently selected from the group consisting of hydrogen, C1-C6 alkyl, and phenyl. n, p, q and r are each independently selected from 0, 1, 2, 3, 4, 5 and 6.

[0007] In some embodiments, the cancer is a B cell cancer, for example, a cancer selected from the group consisting of chronic lymphocytic leukemia, small lymphocytic leukemia, mantle cell lymphoma, non-Hodgkin's lymphoma, marginal zone lymphoma, and Waldenstrom's macroglobulinemia.

[0008] Further disclosed herein is a method of treating a cancer alleviated by selective inhibition of BTK in a patient in need thereof, comprising administering to the patient an effective amount of a tyrosine kinase inhibitor. In some embodiments, the cancer has a BTK mutation, e.g., the cancer has been identified as having a BTK mutation, e.g., a C481 mutation.

[0009] In some embodiments, the tyrosine kinase inhibitor is an inhibitor of BTK. In some embodiments, the BTK inhibitor for use in the methods described herein can be, for example, tert-butyl (4-(4-amino-1-(2-(4-(dimethylamino)piperidin-1-yl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenyl)carbamate or a pharma- ceutical acceptable salt thereof, represented by the formula: TIFF2025514034000003.tif3172

[0010] For example, disclosed herein is a method of treating a cancer harboring a BTK mutation, e.g., a cancer identified as having a BTK mutation, in a patient in need thereof, comprising administering to the patient an effective amount of a tyrosine kinase inhibitor, e.g., tert-butyl (4-(4-amino-1-(2-(4-(dimethylamino)piperidin-1-yl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenyl)carbamate, or a pharma- ceutical acceptable salt thereof.

[0011] Further disclosed herein is a method of treating a B cell cancer harboring a BTK C481 mutation in a patient in need thereof, comprising administering to the patient an effective amount of tert-butyl (4-(4-amino-1-(2-(4-(dimethylamino)piperidin-1-yl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenyl)carbamate, or a pharmaceutically acceptable salt thereof, wherein the cancer is resistant to treatment with ibrutinib, acalabrutinib, zanubrutinib, and / or fenebrutinib.

[0012] Also provided herein is a method of treating squamous cell carcinoma in a patient identified as having carcinoma and in need of treatment, comprising administering to the patient an effective amount of a tyrosine kinase inhibitor having the following formula: TIFF2025514034000004.tif3172 Here, the squamous cell carcinoma is selected from the group consisting of cutaneous squamous cell carcinoma, esophageal squamous cell carcinoma, tongue squamous cell carcinoma, head and neck squamous cell carcinoma, vulvar squamous cell carcinoma, and lung squamous cell carcinoma. [Brief description of the drawings]

[0013] [Figure 1] 1 is a square and dot plot showing significant sensitivity of pan-cancer squamous cell lines to Compound A (p<0.05).

[0014] [Diagram 2] Figure 1 shows the statistical association between gene conversion in 38 cancer genes and a shift in sensitivity to compound A (measured by 10 log IC50). A shift to the left represents increased sensitivity. The size of the circle represents the number of cell lines carrying the alteration (minimum number of cell lines per alteration).

[0015] [Diagram 3] The 10 log IC50 distribution of compound A versus FAT1 mutation status is shown (0=wild type, 1=mutated, p=0.16).

[0016] [Figure 4] Tumor volume in a KYSE70 (esophageal squamous cell carcinoma) xenograft model is shown over time by treatment group (vehicle or Compound A).

[0017] [Diagram 5] Tumor volume in a Cal27 (tongue squamous cell carcinoma) xenograft model over time by treatment group (vehicle or Compound A). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] The features and other details of the present disclosure are described in more detail below.Before further description of the present disclosure, certain terms employed in the present specification, examples and appended claims are collected here.These definitions should be read in the light of the remaining parts of the present disclosure as understood by those skilled in the art.Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.

[0019] definition The term "alkyl" as used herein refers to a saturated straight or branched hydrocarbon, e.g., a straight or branched group of 1-6, 1-5, 1-4, or 1-3 carbon atoms, referred to herein as C1-C6 alkyl, C2-C6 alkyl, C1-C4 alkyl, and C1-C3 alkyl, respectively. For example, "C1-C6 alkyl" refers to a straight or branched saturated hydrocarbon containing 1-6 carbon atoms. Examples of C1-C6 alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, and neopentyl. In another example, "C2-C6 alkyl" refers to a straight or branched saturated hydrocarbon containing 1-5 carbon atoms. Examples of C2-C6 alkyl groups include, but are not limited to, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, and neopentyl. In a further example, "C1-C4 alkyl" refers to a linear or branched saturated hydrocarbon containing 1 to 4 carbon atoms. Examples of C1-C4 alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, isopropyl, isobutyl, sec-butyl, and tert-butyl. Exemplary alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 3-methyl-2-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, and hexyl.

[0020] The term "alkenyl" as used herein refers to an unsaturated straight or branched chain hydrocarbon having at least one carbon-carbon double bond. Exemplary alkenyl groups include, but are not limited to, straight or branched chain groups having 2 to 6 or 3 to 4 carbon atoms, referred to herein as C2-C5 alkenyl, C2-C6 alkenyl, and C3-C4 alkenyl, respectively. Exemplary alkenyl groups include, but are not limited to, vinyl, allyl, butenyl, pentenyl, and the like.

[0021] The term "alkynyl" as used herein refers to an unsaturated straight or branched chain hydrocarbon having at least one carbon-carbon triple bond. Exemplary alkynyl groups include, but are not limited to, straight or branched chain groups having 2 to 6 or 3 to 6 carbon atoms, referred to herein as C2-6 alkynyl and C3-6 alkynyl, respectively. Exemplary alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, methylpropynyl, and the like.

[0022] The term "alkoxy" as used herein refers to a straight or branched alkyl group attached to an oxygen (alkyl-O-). Exemplary alkoxy groups include, but are not limited to, alkoxy groups having 1 to 6 or 2 to 6 carbon atoms, referred to herein as C1-C5 alkoxy, C1-C6 alkoxy, and C2-C6 alkoxy, respectively. Exemplary alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy, and the like.

[0023] The term "cycloalkyl" as used herein refers to a monocyclic saturated or partially unsaturated hydrocarbon ring (carbocyclic) system, e.g., each ring is fully saturated or contains one or more units of unsaturation, but no ring is aromatic. A cycloalkyl can have 3 to 6 or 4 to 6 carbon atoms in its ring system, and is referred to herein as C3-C6 cycloalkyl or C4-C6 cycloalkyl, respectively. Exemplary cycloalkyl groups include, but are not limited to, cyclohexyl, cyclohexenyl, cyclopentyl, cyclopentenyl, cyclobutyl, and cyclopropyl.

[0024] The term "aryl" refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10 or 14 pi electrons shared in a cyclic arrangement) having 6 to 14 ring carbon atoms and zero heteroatoms provided to the aromatic ring system ("C6- 14 In some embodiments, an aryl group has 6 ring carbon atoms ("C6 aryl", e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("C 10 aryl," e.g., naphthyl, such as 1-naphthyl and 2-naphthyl. In some embodiments, an aryl group has 14 ring carbon atoms ("C 14"Aryl" includes ring systems in which an aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups, where the radical or point of attachment is on the aryl ring, and in such cases the number of carbon atoms continues to refer to the number of carbon atoms in the aryl ring system. Typical aryl groups include, but are not limited to, groups derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene, hexalene, as-indacene, s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, preadene, pyrene, pyranthrene, rubicene, triphenylene and trinaphthalene.Specific aryl groups include phenyl, naphthyl, indenyl and tetrahydronaphthyl.

[0025] The terms "halo" and "halogen," as used herein, refer to fluoro (F), chloro (Cl), bromo (Br) and / or iodo (I).

[0026] The terms "hydroxy" and "hydroxyl" as used herein refer to an --OH group.

[0027] The term "hetero", when used to describe a compound or a group present on a compound, means that one or more carbon atoms in the compound or group are replaced with one or more heteroatoms. As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Examples of heteroatoms include, but are not limited to, nitrogen, oxygen, and sulfur.

[0028] Hetero can apply to any of the above hydrocarbyl groups such as alkyl having 1 to 5, especially 1 to 3, heteroatoms, e.g., heteroalkyl; cycloalkyl, e.g., heterocyclyl; aryl, e.g., heteroaryl; cycloalkenyl, e.g., cycloheteroalkenyl.

[0029] The term "heteroaryl" or "heteroaromatic group" as used herein refers to an aromatic 5-10 membered ring system containing one or more heteroatoms, e.g., 1-3 heteroatoms such as nitrogen, oxygen, sulfur, etc. The term can also be used to refer to a 5-7 membered monocyclic heteroaryl or an 8-10 membered bicyclic heteroaryl. Where possible, the heteroaryl ring may be linked to adjacent radicals, but not limited to carbon or nitrogen. Examples of heteroaryl rings include, but are not limited to, furan, thiophene, pyrrole, pyrrolopyridine, indole, thiazole, oxazole, isothiazole, isoxazole, imidazole, benzimidazole, imidazopyridine, pyrazole, triazole, pyridine, or pyrimidine.

[0030] The terms "heterocyclyl", "heterocycle" or "heterocyclic group" are art-recognized and refer to saturated or partially unsaturated 4-10 membered ring structures that contain one to three heteroatoms, such as nitrogen, oxygen, and sulfur. When possible, the heterocyclyl ring may be linked to adjacent radicals through a carbon or nitrogen. The term may also be used to refer to 4-10 membered saturated or partially unsaturated ring structures that are bridged, fused, or spirocyclic ring structures that contain one to three heteroatoms, such as nitrogen, oxygen, and sulfur. Examples of heterocyclyl groups include, but are not limited to, pyrrolidine, piperidine, morpholine, thiomorpholine, piperazine, oxetane, azetidine, tetrahydrofuran, dihydrofuran, dihydropyran, tetrahydropyran, and the like. In some embodiments, the heterocycle is a spiroheterocycle. In some embodiments, the heterocycle is a bridged heterocycle. "Spiroheterocyclyl" or "spiroheterocycle" refers to a polycyclic heterocyclyl having rings joined through one common atom (called a spiroatom), the rings containing N, O, and S(O) as ring atoms. m (wherein m is an integer of 0 to 2).

[0031] "SRC family", "SRC kinase family" and "SRC family of kinases" refer to a family of non-receptor tyrosine kinases including, but not limited to, BLK, FGR, FRK, FYN, HCK, LCK, LYN, SRC and YES.

[0032] "Treatment" includes any effect that results in the improvement of a condition, disease, disorder, etc., e.g., alleviation, reduction, modulation, or elimination. For example, "treating" or "treatment" of a condition, disorder, or condition includes: (1) preventing or delaying the appearance of clinical symptoms of an existing condition, disorder, or condition in a human suffering from or predisposed to suffering from the condition, disorder, or condition, but who has not yet experienced or manifested clinical or subclinical symptoms of the condition, disorder, or condition; (2) inhibiting the condition, disorder, or condition, i.e., arresting, reducing, or delaying the progression of the disease or its recurrence (in the case of maintenance therapy) or at least one clinical or subclinical symptom thereof; or (3) relieving or attenuating the disease, i.e., causing regression of the condition, disorder, or condition, or at least one clinical or subclinical symptom thereof. It is to be understood that "treating" or "treatment" includes prevention as well as alleviation of established symptoms.

[0033] The term "disorder" refers to, and is used interchangeably with, the terms "disease," "condition," or "illness," unless otherwise specified.

[0034] "Pharmaceutically or pharmacologically acceptable" includes molecular entities and compositions that do not produce adverse, allergic or other untoward reactions when administered to animals or humans. For human administration, preparations should meet the standards of sterility, pyrogenicity, general safety and purity as required by FDA biologics standards.

[0035] "Individual", "patient" or "subject" are used interchangeably and include any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, or primates, and most preferably humans. The compounds of the present disclosure may be administered to mammals, such as humans, but also to other mammals, such as animals requiring veterinary treatment, for example, livestock animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, sheep, pigs, horses, etc.) and laboratory animals (e.g., rats, mice, guinea pigs, etc.). The mammal treated in the methods of the present disclosure is desirably a mammal in which treatment of, for example, cancer or a blood disorder is desired. "Modulation" includes antagonism (e.g., inhibition), agonism, partial antagonism and / or partial agonism.

[0036] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a compound of interest that elicits a biological or medical response in a tissue, system or animal (e.g., a mammal or human) that is desired by a researcher, veterinarian, medical doctor or other clinician. The compounds of the present disclosure are administered in a therapeutically effective amount to treat a disease. Alternatively, a therapeutically effective amount of a compound is the amount necessary to achieve a desired therapeutic and / or prophylactic effect.

[0037] As used herein, the term "pharmaceutically acceptable salt" refers to salts of acidic or basic groups that may be present in the compounds used in the compositions. Compounds included in the compositions that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. Acids that can be used to form pharma- ceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, for example, salts containing pharma- ceutically acceptable anions, including, but not limited to, malate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)). The compounds contained in the present composition that are acidic in nature can form base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal salts or alkaline earth metal salts, particularly calcium salts, magnesium salts, sodium salts, lithium salts, zinc salts, potassium salts and iron salts. Compounds contained in the present composition that contain a basic or acidic moiety can also form pharma-ceutically acceptable salts with various amino acids. Compounds of the present disclosure may contain both acidic and basic groups, for example, one amino group and one carboxylic acid group. In such cases, the compounds may exist as acid addition salts, zwitterions or base salts.

[0038] The compounds of the present disclosure may contain one or more chiral centers and / or double bonds and therefore may exist as stereoisomers, such as geometric isomers, enantiomers or diastereomers. As used herein, the term "stereoisomer" includes all geometric isomers, enantiomers and / or diastereomers of a compound. For example, if a compound is depicted with a particular chiral center(s), a compound depicted without such chirality at that and other chiral centers of the compound is within the scope of the present disclosure. That is, a compound depicted in two dimensions with "flat" or "straight" bonds, for example, rather than in three dimensions with solid or dashed wedge bonds. Stereospecific compounds may be designated with the symbols "R" or "S", depending on the arrangement of the substituents around the stereogenic carbon atom. The present disclosure encompasses all the various stereoisomers of these compounds and mixtures thereof. Mixtures of enantiomers or diastereomers may be designated "(±)" in the nomenclature, although one of skill in the art will recognize that structures may implicitly indicate chiral centers. Drawings of chemical structures, e.g., generic chemical structures, are understood to encompass all stereoisomeric forms of the named compound, unless otherwise indicated.

[0039] Individual enantiomers and diastereomers of the compounds of the present disclosure can be prepared synthetically from commercially available starting materials containing asymmetric or stereogenic centers, or by preparing a racemic mixture followed by separation methods well known to those skilled in the art. These separation methods include (1) binding the mixture of enantiomers to a chiral auxiliary, recrystallizing or chromatographically separating the resulting mixture of diastereomers, and liberating the optically pure products from the auxiliary, (2) forming salts with optically active resolving agents, (3) directly separating the mixture of optical enantiomers on a chiral liquid chromatography column, or (4) kinetically separating using stereoselective chemical or enzymatic reagents. Racemic mixtures can also be resolved into the constituent enantiomers by well-known methods, such as chiral phase gas chromatography or crystallization in chiral solvents. Stereoselective syntheses are chemical or enzymatic reactions in which a single reactant forms an unequal mixture of stereoisomers upon the generation of a new stereocenter or the transformation of an existing stereocenter, and are well known in the art. Stereoselective synthesis includes both enantioselective and diastereoselective transformations, see, e.g., Carreira and Kvaerno, Classics in Stereoselective Synthesis, Wiley-VCH:Weinheim, 2009.

[0040] Geometric isomers resulting from the arrangement of substituents around a carbon-carbon double bond or around a cycloalkyl or heterocycloalkyl may also exist in the compounds of the present disclosure. The symbol (double line with dashed line and straight line) indicates a bond that may be a single bond, a double bond, or a triple bond as described herein. Substituents around a carbon-carbon double bond are designated as being in the "Z" or "E" configuration, where the terms "Z" and "E" are used according to IUPAC standards. Unless otherwise specified, structures depicting double bonds encompass both the "E" and "Z" isomers.

[0041] Substituents around a carbon-carbon double bond can be designated "cis" or "trans", with "cis" referring to substituents on the same side of the double bond and "trans" referring to substituents on opposite sides of the double bond. The arrangement of substituents around a carbocyclic ring can also be designated "cis" or "trans". The term "cis" refers to substituents on the same side of the plane of the ring and the term "trans" refers to substituents on opposite sides of the plane of the ring. A mixture of compounds in which the substituents are located on both the same and opposite sides of the plane of the ring is referred to as "cis / trans".

[0042] The compounds disclosed herein can exist in solvated and unsolvated forms with pharma- ceutically acceptable solvents such as water, ethanol, etc., and the present disclosure is intended to encompass both solvated and unsolvated forms. In one embodiment, the compound is amorphous. In one embodiment, the compound is a single polymorph. In another embodiment, the compound is a mixture of polymorphs. In another embodiment, the compound is in a crystalline form.

[0043] The present disclosure also encompasses isotopically labeled compounds of the present disclosure that are identical to those described herein, except that one or more atoms are replaced with an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, e.g., 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F and 36 For example, the compounds of the present disclosure may have one or more H atoms replaced with deuterium.

[0044] Certain isotopically labeled disclosed compounds (e.g., 3 H and 14C) are useful in compound and / or substrate tissue distribution assays. Tritium isotopes (i.e., 3 H) and carbon-14 isotopes (i.e., 14 C) is particularly preferred because it is easy to prepare and detect. 2 Substitution with heavy isotopes such as H) may confer certain therapeutic advantages resulting from greater metabolic stability (e.g., increased half-life in vivo or reduced dosage requirements) and therefore may be preferred in some circumstances. Isotopically labeled compounds of the present disclosure can generally be prepared following procedures similar to those disclosed in the Examples herein by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0045] In this disclosure, the term "and / or" is used to mean either "and" or "or," unless otherwise indicated.

[0046] As used herein, the terms "a" and "an" are intended to mean one or more, unless otherwise specified. For example, the term "agent" includes both a single agent and a combination of two or more agents.

[0047] When the term "about" is used before a quantitative value, the present disclosure also includes the specific quantitative value itself, unless specifically stated otherwise. As used herein, the term "about" refers to a ±10% variation from the nominal value, unless otherwise indicated or inferred.

[0048] method In some embodiments, disclosed herein is a method of treating cancer in a patient in need thereof, comprising administering to the patient an effective amount of a Bruton's tyrosine kinase (BTK) inhibitor represented by Formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof. TIFF2025514034000005.tif3285In formula, R 1 NHR a and N.R. a Rb is selected from the group consisting of: R a and R b are each independently selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C3-C6 alkynyl, C3-C6 cycloalkyl, and C3-C6 cycloalkenyl, or R a and R b may be taken together with the nitrogen to which they are attached to form a C3-C6 heterocyclyl. R 2 OR 11 , Hydrogen, Halo, NHR 11 , C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl. R 3 is NHCO2R 4 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, aryl, halo, aryloxy, NH(CO)NR 5 R 6 , NH(CO)R 7 , NH-C1-C6 alkyl, NH-C2-C6 alkenyl, NH(CH2) n -aryl, (CH2) p -heteroaryl, (CH2) q CO2R 8 , (CH2) r COR 9 and NHSO2R 10 wherein each C1-C6 alkyl, C2-C6 alkenyl, aryl or heteroaryl group in the preceding list is independently selected from the group consisting of C1-C6 alkyl, halo, OH, NR c R d ,CONR c R d , C1-C6 alkoxy, aryloxy, and CO2H. R 4 From R 11 are each independently selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, and aryl. R c and Rd are each independently selected from the group consisting of hydrogen, C1-C6 alkyl, and phenyl. n, p, q and r are each independently selected from 0, 1, 2, 3, 4, 5 and 6.

[0049] In some embodiments, R a and R b is, each independently, C1-C6 alkyl or C2-C6 alkenyl. For example, in certain embodiments, R a and R b Each is CH. In another embodiment, R a is C1-C6 alkyl or C2-C6 alkenyl, R b is hydrogen. For example, in certain embodiments, R a is CH3 and R b is hydrogen.

[0050] In a further embodiment, R 2 is C1-C6 alkoxy or hydrogen. For example, in certain embodiments, R 2 is OCH3. In another embodiment, R 4 From R 11 are each independently C1-C6 alkyl.

[0051] In some embodiments, R 3 is NHCO2-C1-C6 alkyl, NHCO-C1-C6 alkyl, NH(CH2) n -aryl, NHCONH-C1-C6 alkyl, (CH2) p -heteroaryl and (CH2) q For example, in certain embodiments, R 3 is selected from the group consisting of NHCO2-tBu, NHCOCH2C(CH3)3, NHCH2phenyl, NHCONH-tBu, CH2-(4-methyl-oxazol-2-yl) and CH2CO2-tBu. 3 is NHCO2-tBu.

[0052] In some embodiments, a BTK inhibitor for use in the methods described herein can be, for example, tert-butyl (4-(4-amino-1-(2-(4-(dimethylamino)piperidin-1-yl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenyl)carbamate, or a pharma- ceutically acceptable salt thereof, represented below: TIFF2025514034000006.tif3172

[0053] For example, in certain embodiments, a BTK inhibitor for use in the methods described herein is: TIFF2025514034000007.tif3172

[0054] In other embodiments, the BTK inhibitor for use in the methods described herein can be, for example, tert-butyl (4-(4-amino-1-(2-(4-(methylamino)piperidin-1-yl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenyl)carbamate, or a pharma- ceutically acceptable salt thereof, represented below: TIFF2025514034000008.tif3172

[0055] For example, in certain embodiments, a BTK inhibitor for use in the methods described herein is: TIFF2025514034000009.tif3172

[0056] In some embodiments, the cancer is a B cell cancer, for example, in some embodiments, the cancer is selected from the group consisting of chronic lymphocytic leukemia, small lymphocytic leukemia, mantle cell lymphoma, non-Hodgkin's lymphoma, marginal zone lymphoma, and Waldenstrom's macroglobulinemia.

[0057] In other embodiments, the cancer harbors a BTK mutation. For example, in some embodiments, the cancer has been identified as having a BTK mutation. In certain embodiments, the BTK mutation is a C481 mutation. For example, in certain embodiments, the BTK mutation is selected from the group consisting of, but not limited to, a C481F mutation, a C481G mutation, a C481R mutation, a C481S mutation, and a C481Y mutation. In some embodiments, the BTK mutation is a C481S mutation.

[0058] In some embodiments, the BTK mutation in the cancer is the result of previously treating the cancer with one or more other cancer therapeutic agents. In certain embodiments, treating the cancer with one or more other cancer therapeutic agents is no longer effective in treating the cancer. In other embodiments, the one or more other cancer therapeutic agents are selected from the group consisting of, for example, but not limited to, ibrutinib, acalabrutinib, zanubrutinib, fenebrutinib, tirabrutinib, trebrutinib, evobrutinib, pirtobrutinib, and spebrutinib.

[0059] In some embodiments, the BTK mutation is a heterozygous BTK mutation. In other embodiments, the BTK mutation is a homozygous BTK mutation. In further embodiments, the BTK inhibitor is administered orally. In other embodiments, the BTK inhibitor is administered subcutaneously. In further embodiments, the compound is administered intraperitoneally. In yet other embodiments, the BTK inhibitor is administered intravenously.

[0060] In some embodiments, the method may further optionally include administering one or more additional cancer chemotherapeutic agents. For example, in other embodiments, the method may further optionally include administering an additional cancer chemotherapeutic agent.

[0061] In certain embodiments, the tyrosine kinase inhibitor is an inhibitor of BTK, hi other embodiments, the tyrosine kinase inhibitor further inhibits a member of the SRC kinase family, including, but not limited to, BLK, FGR, FRK, FYN, HCK, LCK, LYN, SRC, and YES.

[0062] Also disclosed herein is a method of treating a cancer alleviated by selective inhibition of BTK in a patient in need thereof, comprising administering to the patient an effective amount of a tyrosine kinase inhibitor represented by the following, or a pharma- ceutical acceptable salt thereof: TIFF2025514034000010.tif3172

[0063] In some embodiments, the cancer is selected from the group consisting of chronic lymphocytic leukemia, small lymphocytic leukemia, mantle cell lymphoma, non-Hodgkin's lymphoma, marginal zone lymphoma, and Waldenstrom's macroglobulinemia. In certain embodiments, the cancer harbors a BTK mutation. For example, in some embodiments, the BTK mutation is a C481 mutation, e.g., a BTK mutation selected from the group consisting of a C481F mutation, a C481G mutation, a C481R mutation, a C481S mutation, and a C481Y mutation.

[0064] Further disclosed herein is a method of treating a cancer harboring a BTK mutation in a patient in need thereof, comprising administering to the patient an effective amount of a tyrosine kinase inhibitor represented by the following formula: TIFF2025514034000011.tif3172

[0065] In some embodiments, the cancer is selected from the group consisting of chronic lymphocytic leukemia, small lymphocytic leukemia, mantle cell lymphoma, non-Hodgkin's lymphoma, marginal zone lymphoma, and Waldenstrom's macroglobulinemia. In certain embodiments, the cancer harbors a BTK mutation. For example, in some embodiments, the BTK mutation is a C481 mutation, e.g., a BTK mutation selected from the group consisting of a C481F mutation, a C481G mutation, a C481R mutation, a C481S mutation, and a C481Y mutation.

[0066] In some embodiments, the BTK mutation in the cancer is the result of previously treating the cancer with one or more other cancer therapeutic agents. In certain embodiments, treating the cancer with one or more other cancer therapeutic agents is no longer effective in treating the cancer. In other embodiments, the one or more other cancer therapeutic agents are selected from the group consisting of, for example, but not limited to, ibrutinib, acalabrutinib, zanubrutinib, fenebrutinib, tirabrutinib, trebrutinib, evobrutinib, pirtobrutinib, and spebrutinib.

[0067] Further disclosed herein is a method of treating a B cell cancer harboring a BTK C481 mutation in a patient in need thereof, comprising administering to the patient an effective amount of tert-butyl (4-(4-amino-1-(2-(4-(dimethylamino)piperidin-1-yl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenyl)carbamate, or a pharmaceutically acceptable salt thereof, wherein the cancer is resistant to treatment with ibrutinib, acalabrutinib, zanubrutinib, fenebrutinib, tirabrutinib, trebrutinib, evobrutinib, pirtobrutinib, and / or spebrutinib.

[0068] In some embodiments, the cancer types disclosed herein, such as B cell cancers, have been identified as having BTK mutations. Methodologies for detecting BTK mutations contemplated herein are known in the art.

[0069] In particular, in certain embodiments, the present disclosure provides a method of treating the above medical indications comprising administering to a patient in need thereof an effective amount of a BTK inhibitor disclosed herein. In certain other embodiments, the present disclosure provides a method of treating the above medical indications in a patient in need thereof comprising orally, subcutaneously, or intravenously administering to the patient a composition comprising a disclosed BTK inhibitor.

[0070] Without wishing to be bound by any particular theory, it is believed that the compounds of the present disclosure (e.g., tert-butyl (4-(4-amino-1-(2-(4-(dimethylamino)piperidin-1-yl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenyl)carbamate) are also inhibitors of SRC family kinases, and that their combined effect on both SRC family kinases and BTK (both wild-type and mutant) is advantageous in the treatment of B cell cancers.

[0071] In some embodiments, the therapeutic methods disclosed herein may provide an anti-proliferative effect. In some embodiments, the therapeutic methods disclosed herein may be methods for treating a proliferative disease or disorder. The terms "proliferative disease" and "proliferative disorder" are used interchangeably herein and relate to unwanted, excessive or abnormal cell proliferation, e.g., undesired or uncontrolled cell proliferation, such as neoplastic or hyperplastic proliferation, whether in vitro or in vivo. Examples of proliferative conditions include, but are not limited to, pre-malignant and malignant cell proliferation, including, but not limited to, cancer, lymphoma, leukemia, solid tumors.

[0072] Further disclosed herein is a method of treating squamous cell carcinoma in a patient identified as having said carcinoma and in need of treatment, comprising administering to the patient an effective amount of a tyrosine kinase inhibitor of Formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof. TIFF2025514034000012.tif3282In formula, R 1 NHR a and N.R. a R b is selected from the group consisting of: R a and R b are each independently selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C3-C6 alkynyl, C3-C6 cycloalkyl, and C3-C6 cycloalkenyl, or R a and R b may be taken together with the nitrogen to which they are attached to form a C3-C6 heterocyclyl. R 2 OR 11 , Hydrogen, Halo, NHR 11 , C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl. R 3 is NHCO2R 4 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, aryl, halo, aryloxy, NH(CO)NR 5 R 6 , NH(CO)R 7 , NH-C1-C6 alkyl, NH-C2-C6 alkenyl, NH(CH2) n -aryl, (CH2) p -heteroaryl, (CH2) q CO2R 8 , (CH2) r COR 9 and NHSO2R 10 wherein each C1-C6 alkyl, C2-C6 alkenyl, aryl or heteroaryl group in the preceding list is independently selected from the group consisting of C1-C6 alkyl, halo, OH, NR c R d ,CONR c R d , C2-C6 alkoxy, aryloxy, and CO2H. R 4 From R 11are each independently selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, and aryl. R c and R d are each independently selected from the group consisting of hydrogen, C1-C6 alkyl, and phenyl. n, p, q and r are each independently selected from 0, 1, 2, 3, 4, 5 and 6.

[0073] In some embodiments, R a and R b is, each independently, C1-C6 alkyl or C2-C6 alkenyl. For example, in certain embodiments, R a and R b Each is CH. In another embodiment, R a is C1-C6 alkyl or C2-C6 alkenyl, R b is hydrogen. For example, in certain embodiments, R a is CH3 and Rb is hydrogen.

[0074] In a further embodiment, R 2 is C1-C6 alkoxy or hydrogen. For example, in certain embodiments, R 2 is OCH3. In another embodiment, R 4 From R 11 are each independently C1-C6 alkyl.

[0075] In some embodiments, R 3 is NHCO2-C1-C6 alkyl, NHCO-C1-C6 alkyl, NH(CH2) n -aryl, NHCONH-C1-C6 alkyl, (CH2) p -heteroaryl and (CH2) q For example, in certain embodiments, R 3is selected from the group consisting of NHCO2-tBu, NHCOCH2C(CH3)3, NHCH2phenyl, NHCONH-tBu, CH2-(4-methyl-oxazol-2-yl) and CH2CO2-tBu. 3 is NHCO2-tBu.

[0076] For example, in some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of: TIFF2025514034000013.tif31156

[0077] In certain embodiments, the tyrosine kinase inhibitor is: TIFF2025514034000014.tif3172

[0078] In some embodiments, the squamous cell carcinoma is, for example, selected from the group consisting of cutaneous squamous cell carcinoma, esophageal squamous cell carcinoma, tongue squamous cell carcinoma, head and neck squamous cell carcinoma, vulvar squamous cell carcinoma, and lung squamous cell carcinoma.

[0079] For example, provided herein is a method of treating squamous cell carcinoma in a patient identified as having said carcinoma and in need of treatment, comprising administering to the patient an effective amount of a tyrosine kinase inhibitor represented by the following formula, or a pharma- ceutical acceptable salt thereof: TIFF2025514034000015.tif3172 Here, the squamous cell carcinoma is selected from the group consisting of cutaneous squamous cell carcinoma, esophageal squamous cell carcinoma, tongue squamous cell carcinoma, head and neck squamous cell carcinoma, vulvar squamous cell carcinoma, and lung squamous cell carcinoma.

[0080] The treatment methods defined herein can be applied as monotherapy or can be combined with conventional surgery, radiation therapy, gene therapy, or chemotherapy or molecular targeted drug therapy in addition to the compounds of the present invention. Such combined treatment can be achieved by simultaneous, sequential, or separate administration of the individual components of the treatment. The term "combination" as used herein refers to simultaneous, separate, or sequential administration. In some embodiments, "combination" refers to simultaneous administration. In other embodiments, "combination" refers to separate administration. In further embodiments, "combination" refers to sequential administration. When administration is sequential or separate, the delay in administration of the second component should not be such that the beneficial effect of the combination is lost. EXAMPLES

[0081] The following non-limiting examples illustrate the present disclosure.

[0082] tert-Butyl (4-(4-amino-1-(2-(4-(dimethylamino)piperidin-1-yl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenyl)carbamate (compound A) can be prepared according to the synthetic procedures described in WO2016 / 185160, the contents of which are incorporated herein by reference. The desmethyl derivative tert-butyl (4-(4-amino-1-(2-(4-(methylamino)piperidin-1-yl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-methoxyphenyl)carbamate (compound B) can be prepared by a similar synthetic procedure.

[0083] Example 1 Tyrosine kinase inhibition test Compound A was dissolved in dimethyl sulfoxide (DMSO) and further diluted with assay buffer to a final concentration of 0.5 μmol / L. A reference compound for assay control was also prepared in the same manner. For BTK inhibition tests, full-length human BTK [2-659 (terminal) amino acids of accession number NP_000052] was expressed as an N-terminal GST fusion protein (103 kDa) using a baculovirus expression system. GST-BTK was purified using glutathione sepharose chromatography. For BTK[C481S] inhibition tests, full-length human BTK [2-659 (terminal) amino acids and C481S of accession number NP_000052] was expressed as an N-terminal GST fusion protein (103 kDa) using a baculovirus expression system. GST-BTK[C481S] was purified using glutathione sepharose chromatography.

[0084] A 4x substrate / ATP / metal solution was prepared in buffer (20mmol / L HEPES, 0.01% TritonX-100, 5mmol / L DTT, pH 7.5), and a 2x kinase solution was also prepared in buffer (20mmol / L HEPES, 0.01% TritonX-100, 1mmol / L DTT, pH 7.5). 5μL of 4x compound solution, 5μL of 4x substrate / ATP / metal solution, and 10μL of 2x kinase solution were mixed and incubated at room temperature for 1 hour in a well of a polypropylene 384-well microplate. 70μL of termination buffer (QuickScout Screening Assist MSA) was added to the well. The reaction mixture was applied to the LabChipTM system, and the peaks of the product and substrate peptide were separated and quantified. The kinase reaction was evaluated by the product ratio (P / (P+S)) calculated from the peak heights of the product (P) and substrate (S) peptide. [Table 1]

[0085] The inhibition rate of each test solution was calculated based on the readout value of the reaction control (complete reaction mixture) as 0% inhibition rate and the readout value of the background (enzyme (-)) as 100% inhibition rate (Table 2). [Table 2]

[0086] Example 2 Pan-cancer squamous cell carcinomas showed high sensitivity to Compound A. For example, Figure 1 shows a square and dot plot showing significant sensitivity (p<0.05) of pan-cancer squamous cell lines to Compound A. Examples of squamous cell carcinomas contemplated herein include, but are not limited to, cutaneous squamous cell carcinoma, esophageal squamous cell carcinoma, head and neck squamous cell carcinoma, vulvar squamous cell carcinoma, and lung squamous cell carcinoma.

[0087] To identify genetic drivers of sensitivity, transformations in 38 cancer genes were observed to be statistically associated with shifts in compound sensitivity in a panel of 102 cell lines, demonstrating clear trends. For example, Figure 2 shows the correlation between gene transformations in 38 cancer genes and compound A ( 10 logIC 50 Figure 1 shows the statistical association between the shift in sensitivity to the alteration (measured by α, β, β-glucose, β-glucose, β-glucose, and β-glucose). A shift to the left represents increased sensitivity. The size of the circle represents the number of cell lines carrying the alteration (minimum number of cell lines per alteration).

[0088] Without wishing to be bound by any particular theory, gene conversion of the FAT1 gene is believed to be highly prevalent in squamous cell carcinoma. For example, FAT1 is altered (mutation of known significance) in 3% of patients across all cancer types, and is highly prevalent in squamous cell carcinoma, particularly skin cancer and head and neck cancer. Figure 3 shows the relationship between compound A and the mutation state of FAT1. 10 logIC 50 Distributions are shown (0=wild type, 1=mutant, p=0.16).

[0089] Example 3 The purpose of this study was to examine the antitumor effect of oral administration of compound A using a KYSE70 mouse xenograft model of esophageal squamous cell carcinoma.

[0090] overview Prior to the in-life phase of the study, animals were weighed and examined for signs of disability. Animals deemed suitable were randomly assigned to one of two treatment groups: Compound A (40 mg / kg) or vehicle control. Compound A or vehicle was administered orally once daily for 28 days. Tumor readings were collected every 2 days up to 28 days, and tumor volumes were adjusted to 100 mm 3 Treatment was started at 2000 mm 3 Treatment was terminated at time t. A t-test was applied to determine the significance of differences in tumor volume.

[0091] Protocol Compound A was dissolved at 4 mg / mL in vehicle (3 mmol / L sodium citrate buffer, pH 3.0) and administered orally once daily (100 μL per 10 g of mouse body weight). The administration material was aliquoted weekly and stored frozen at −20° C. until administration. To enroll mice in the study, the minimum palpable tumor volume (∼100 mm measured by caliper) was measured in the required number of mice. 3 Subcutaneous tumor volumes were monitored daily until tumor volume reached 0.001 mg / kg / day. Tumor-bearing mice were randomized and assigned to treatment cohorts, resulting in comparable mean tumor volumes and statistics. After randomization and cohort assignment, each treatment cohort was treated according to the dose and schedule. Mice were euthanized when they reached the study endpoint.

[0092] Tumor volumes were measured every 2 days (maximum 3 times a week) using calibrated calipers and were calculated using the formula: length (mm) × width (mm) × height (mm) × 0.50.

[0093] The humane endpoint for tumor volume was when a 20% loss in body weight (excluding tumor weight) was observed in 1 week, or a 15% loss in body weight was observed in 2–3 days, or a body condition score of less than 2 (body condition scoring for scoring guide), or tumor weight exceeded 10% of tumor-free body weight (tumor weight is tumor height × length × width × 0.50 × density 1 g / cm). 3 (based on the calculation) or tumor size is 2000 mm 3 Body weight measurements were performed every 2 days (maximum 3 times per week) during the 28-day treatment period in accordance with the tumor volume measurement schedule. The humane endpoint was reached when body weight loss exceeded 20%.

[0094] result Treatment with Compound A significantly inhibited tumor growth. The mean tumor volume at baseline was 101.4 mm in the vehicle group. 3 , 109.8 mm for compound A group 3 The mean tumor volume on day 27 was 580.5 mm in the vehicle group. 3 , and 31.5 mm in compound A group. 3 The vehicle control group had a 472% increase in tumor volume, while the compound A group had a 71% decrease in tumor volume. By Student's t-test, the tumor volume on day 27 was significantly lower in the compound A group than in the vehicle control group (p≦0.001). The mean baseline body weight was 24.3 g in the compound A group and 24.5 g in the vehicle control group. The mean increase in body weight from baseline on day 27 was 2.0% in the compound A 40 mg / kg group and 3.5% in the vehicle control group.

[0095] A representation of tumor volume over time is shown in Figure 4. Table 3 shows the relative tumor volume and tumor growth inhibition at day 27 by treatment group in the KYSE70 (esophageal squamous cell carcinoma) xenograft model. Abbreviated RTV28 (relative tumor volume, day 28) = [mean tumor volume D28] / [mean tumor volume at baseline]. Abbreviated TGI% (percent tumor growth inhibition) = (1 - [RTV28 of treatment group] / [RTV28 vehicle]) x 100. [Table 3]

[0096] Example 4 The purpose of this study was to investigate the antitumor effect of oral administration of compound A using a tongue squamous cell carcinoma Cal27 mouse xenograft model.

[0097] overview Prior to the in-life phase of the study, animals were weighed and examined for signs of disability. Animals deemed suitable were randomly assigned to one of two treatment groups: Compound A (40 mg / kg) or vehicle control. Compound A or vehicle was administered orally once daily for 28 days. Tumor readings were collected every 2 days up to 28 days, and tumor volumes were adjusted to 100 mm 3 Treatment was started at 2000 mm 3 Treatment was terminated at time t. A t-test was applied to determine the significance of differences in tumor volume.

[0098] Protocol Compound A was dissolved at 4 mg / mL in vehicle (3 mmol / L sodium citrate buffer, pH 3.0) and administered orally once daily (100 μL per 10 g of mouse body weight). The administration material was aliquoted weekly and stored frozen at −20° C. until administration. To enroll mice in the study, the minimum palpable tumor volume (∼100 mm measured by caliper) was measured in the required number of mice. 3 Subcutaneous tumor volumes were monitored daily until tumor volume reached 0.001 mg / kg / day. Tumor-bearing mice were randomized and assigned to treatment cohorts, resulting in comparable mean tumor volumes and statistics. After randomization and cohort assignment, each treatment cohort was treated according to the dose and schedule. Mice were euthanized when they reached the study endpoint.

[0099] Tumor volumes were measured every 2 days (maximum 3 times a week) using calibrated calipers and were calculated using the formula: length (mm) × width (mm) × height (mm) × 0.50.

[0100] The humane endpoint for tumor volume was when a 20% decrease in body weight (excluding tumor weight) was observed over a 1-week period, or a 15% decrease in body weight was observed over a 2-3 day period, or a body condition score of less than 2 (Body Condition Scoring for Scoring Guide), or tumor weight exceeded 10% of tumor-free body weight (tumor weight is calculated as tumor height x length x width x 0.50 x density 1 g / cm). 3 (based on the calculation) or tumor size is 2000 mm 3 Body weight measurements were performed every 2 days (maximum 3 times per week) during the 28-day treatment period in accordance with the tumor volume measurement schedule. The humane endpoint was reached when body weight loss exceeded 20%.

[0101] result Administration of Compound A resulted in substantial TGI. The mean tumor volume at baseline was 116.1 mm in the vehicle group. 3 , Compound A group 116.2mm 3 The mean tumor volume on day 27 was 647.2 mm in the vehicle group. 3 , compound A group 91.4mm 3 The tumor volume increased by 457% in the vehicle control group and decreased by 78% in the compound A group. The tumor volume on day 27 was significantly lower in the compound A group than in the vehicle control group by Student's t-test (p≦0.001). The mean baseline body weight was 23.3 g in the compound A group and 23.7 g in the vehicle control group. The mean increase in body weight from baseline on day 27 was 2.1% in the compound A 40 mg / kg group and 1.8% in the vehicle control group.

[0102] A representation of tumor volume over time is shown in Figure 5. Table 4 shows the relative tumor volume and tumor growth inhibition at day 27 by treatment group in the Cal27 (tongue squamous cell carcinoma) xenograft model. Abbreviated RTV28 (relative tumor volume, day 28) = [mean tumor volume D28] / [mean tumor volume at baseline]. Abbreviated TGI% (percent tumor growth inhibition) = (1 - [RTV28 of treatment group] / [RTV28 vehicle]) x 100. [Table 4]

[0103] Incorporation by Reference All publications and patents mentioned in this specification, including those listed below, are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In the case of conflict, the present application, including definitions herein, will control. Equivalent While specific embodiments of the subject disclosure have been described, the above specification is illustrative and not restrictive. Many variations of the present disclosure will become apparent to those skilled in the art upon review of this specification. The full scope of the present disclosure, along with such variations, along with the full scope of equivalents, should be determined by reference to the claims and the specification. Unless otherwise indicated, all numerical values ​​expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are understood to be modified in all instances by the term "about." Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and appended claims are approximations that may vary depending upon the desired properties sought to be obtained by the present disclosure.

Claims

1. A pharmaceutical composition for treating B-cell cancer harboring the BTK C481S mutation in patients in need, comprising an effective amount of a compound selected from the following group or a pharmaceutically acceptable salt or stereoisomer thereof.

2. The pharmaceutical composition according to claim 1, wherein the compound is as follows.

3. The pharmaceutical composition according to claim 1, wherein the compound is as follows.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the cancer is selected from the group consisting of chronic lymphocytic leukemia, small lymphocytic leukemia, mantle cell lymphoma, non-Hodgkin lymphoma, marginal zone lymphoma, and Waldenström macroglobulinemia.

5. The pharmaceutical composition according to claim 1, wherein the BTK mutation in the cancer is a result of having previously treated the cancer with one or more other cancer treatment agents.

6. The pharmaceutical composition according to claim 5, wherein treating cancer with one or more other cancer treatment agents is no longer effective in treating cancer.

7. The pharmaceutical composition according to claim 5 or 6, wherein one or more other cancer treatment agents are selected from the group consisting of ibrutinib, acalabrutinib, zanubrutinib, fenebrutinib, tirabrutinib, trebrutinib, evobrutinib, pirtobrutinib, and spebratinib.

8. The pharmaceutical composition according to claim 1, wherein the BTK mutation is a heterozygous BTK mutation.

9. The pharmaceutical composition according to claim 1, wherein the BTK mutation is a homozygous BTK mutation.

10. The pharmaceutical composition according to claim 1, wherein the compound is administered orally, subcutaneously, intraperitoneally, or intravenously.

11. Furthermore, the pharmaceutical composition according to claim 1, for optionally administering one or more cancer chemotherapy agents.

12. Furthermore, the pharmaceutical composition according to claim 1, for optionally administering one additional cancer chemotherapy agent.

13. The pharmaceutical composition according to claim 1, wherein the compound further inhibits SRC family kinases.

14. A pharmaceutical composition for treating cancer harboring the BTK C481S mutation, which is mitigated by selective inhibition of BTK in patients requiring the treatment, comprising an effective amount of the compound or a pharmaceutically acceptable salt thereof as described below.

15. The pharmaceutical composition according to claim 14, wherein the cancer is selected from the group consisting of chronic lymphocytic leukemia, small lymphocytic leukemia, mantle cell lymphoma, non-Hodgkin lymphoma, marginal zone lymphoma, and Waldenström macroglobulinemia.

16. A pharmaceutical composition for treating cancer harboring the BTK C481S mutation in patients requiring treatment, comprising an effective amount of a compound represented by the following formula or a pharmaceutically acceptable salt thereof, wherein the cancer is selected from the group consisting of chronic lymphocytic leukemia, small lymphocytic lymphoma, mantle cell lymphoma, non-Hodgkin lymphoma, marginal zone lymphoma, and Waldenström macroglobulinemia.

17. The pharmaceutical composition according to claim 16, wherein the BTK mutation in the cancer is a result of having previously treated the cancer with one or more other cancer treatment agents.

18. The pharmaceutical composition according to claim 17, wherein one or more other cancer treatment agents are selected from the group consisting of ibrutinib, acalabrutinib, zanubrutinib, fenebrutinib, tirabrutinib, trebrutinib, evobrutinib, pirtobrutinib, and spebratinib.

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