NEK7 kinase inhibitors

Abstract

Compounds are provided that have activity as inhibitors of NEK7, the compounds having the following structure (I): [Formula 1] TIFF2023528659000213.tif5259, or a pharmaceutically acceptable salt, stereoisomer or prodrug thereof, wherein A, X, Y, R 1 , R 2 , R 3 and R 4 is as defined herein. Methods relating to the preparation and use of such compounds, pharmaceutical compositions comprising such compounds, and methods of modulating NLRP3 inflammasome activity are also provided.

Description

[Technical Field]

[0001] Embodiments of this disclosure generally relate to compounds, as well as methods for preparing and using them as therapeutic or prophylactic agents (for example, for the treatment of inflammation). [Background technology]

[0002] Description of related technologies Inflammasomes are multiprotein complexes whose activation plays a central role in innate immunity and inflammation. To date, four inflammasomes have been described (NLRP1, NLRC4, NLRP3, and AIM2). The NLRP3 inflammasome consists of NLRP3, ASC, and caspase-1. Its activation leads to the activation of caspase-1, which promotes the secretion of IL-1β and IL-18, inflammation-mediating cytokines in animal disease models of several autoimmune diseases, myocardial infarction, metabolic syndrome, inflammatory bowel disease, and macrophage activation syndrome.

[0003] NEK7 is a member of the NIMA-associated kinase (NEK) family, acting as an NLRP3-binding protein and regulating its oligomerization and activation. NEK7 is a serine / threonine kinase essential for mitotic initiation, cell cycle progression, cell division, and mitotic progression. It is expressed in various tissues, including the brain, heart, lungs, liver, and spleen. Overexpression of NEK7 induces the production of abnormal cells and is closely associated with tumors such as retinoblastoma, gallbladder cancer, and head and neck cancers.

[0004] Many inhibitors are widely used to inhibit effector signaling pathways such as IL-1β or IL-18 without eliminating the inflammatory response. Inhibitors of NLRP3 inflammasome activation that block the NLRP3-NEK7 interaction may show therapeutic or prophylactic activity in several human diseases, including type 2 diabetes (T2D), atherosclerosis, gout, and neurodegenerative diseases. However, the exact mechanism of the NLRP-3-NEK7 interaction is not well understood.

[0005] Therefore, there is a need to develop inhibitors that directly target NEK7 and affect the inflammatory response regulated by the NLRP3 inflammasome in several pathological diseases, such as gout, atherosclerosis, type 2 diabetes, metabolic syndrome, macular degeneration, Alzheimer's disease, multiple sclerosis, and inflammatory bowel disease. Embodiments of this disclosure satisfy this need and provide further relevant advantages. [Overview of the project]

[0006] In short, embodiments of the present disclosure provide compounds (including pharmaceutically acceptable salts, stereoisomers, and prodrugs) that can modulate the activity of the NLRP3 inflammasome.

[0007] In one embodiment, the present invention has the following structure (I): [ka] [In the formula, A, X, Y, R 1 , R 2 , R 3 and R 4 Each of these is defined as follows: The present invention provides compounds having [a certain characteristic], pharmaceutically acceptable salts thereof, stereoisomers, or prodrugs.

[0008] In another embodiment, a pharmaceutical composition comprising the disclosed compound and a method of using it for the treatment of inflammation are also provided. [Modes for carrying out the invention]

[0009] In the following description, certain specific details are provided to give a full understanding of the various embodiments of this disclosure. However, those skilled in the art will understand that this disclosure may be implemented without these details.

[0010] Unless the context requires otherwise, the word “comprise” and its variations (such as “comprises” and “comprising”) throughout this specification and the claims should be interpreted in an open and inclusive sense, that is, “including, but not limited to.”

[0011] In this description, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer value, where appropriate, and fractional values ​​(e.g., one-tenth and one-hundredth of an integer) within the listed range, unless otherwise specified. The terms “about” and “approximately” as used herein mean ±20%, ±10%, ±5%, or ±1% of the indicated range, value, or structure, unless otherwise specified. The terms “one (a)” and “one (an)” as used herein should be understood to mean “one or more” of the listed components. The use of options (e.g., “or”) should be understood to mean one of the options, both, or any combination thereof.

[0012] Throughout this specification, any reference to “one embodiment” or “an embodiment” means that a particular feature, structure, or characteristic described in relation to an embodiment is included in at least one embodiment of the present invention. Therefore, the phrases “in one embodiment” or “in an embodiment” appearing in various places throughout this specification do not necessarily all refer to the same embodiment. Furthermore, certain features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0013] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by a person of ordinary skill in the art to which this disclosure pertains. The singular forms “a,” “an,” and “the” used herein and in the claims include plural references unless the context clearly indicates otherwise.

[0014] "Amino" refers to the -NH2 radical.

[0015] "Carboxy" or "carboxyl" refers to the -CO2H radical.

[0016] "Cyano" refers to the -CN radical.

[0017] "Hydroxy" or "hydroxyl" refers to the -OH radical.

[0018] "Nitro" refers to the -NO2 radical.

[0019] "Oxo" refers to an =O substituent.

[0020] "Thiol" refers to the -SH substituent.

[0021] "Thioxo" refers to the =S substituent.

[0022] "Alkyl" consists only of carbon and hydrogen atoms, with 1 to 12 carbon atoms (C1-C1). 12 A saturated linear or branched hydrocarbon radical has 1 to 8 carbon atoms (C1-C8 alkyl) or 1 to 6 carbon atoms (C1-C6 alkyl), or any value within these ranges (e.g., C4-C6 alkyl and the like), and is bonded to the rest of the molecule by a single bond (e.g., methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl and the like). The number of carbon atoms mentioned relates to the carbon skeleton and carbon branching, but does not include carbon atoms of any substituent. Unless otherwise explicitly stated herein, alkyl groups may be substituted as appropriate.

[0023] "Alkenyl" consists only of carbon and hydrogen atoms, has one or more carbon-carbon double bonds, and has 2 to 12 carbon atoms (C2-C 12 Alkenyls refer to unsaturated linear or branched hydrocarbon radicals having 2 to 8 carbon atoms (C2-C8 alkenyls) or 2 to 6 carbon atoms (C2-C6 alkenyls), or any value within these ranges, bonded to the rest of the molecule by a single bond (e.g., ethenyl, propa-1-enyl, buta-1-enyl, penta-1-enyl, penta-1,4-dienyl, and similar groups). The number of carbons mentioned relates to the carbon skeleton and carbon branching, but does not include carbon atoms of any substituents. Unless otherwise explicitly stated herein, alkenyl groups may be substituted as appropriate.

[0024] The term "alkynyl" refers to a group of 2 to 12 carbon atoms (C2-C2). 12"(alkynyl), having from 2 to 9 carbon atoms (C2-C9 alkynyl), or from 2 to 6 carbon atoms (C2-C6 alkynyl), or any value within these ranges, and refers to an unsaturated straight-chain or branched-chain hydrocarbon radical having at least one carbon-carbon triple bond. Examples of alkynyl groups may be selected from groups consisting of ethynyl, propargyl, but-1-ynyl, but-2-ynyl and the like. The number of carbons mentioned relates to the carbon skeleton and carbon branching, and does not include carbon atoms of any substituents. Unless otherwise clearly stated herein, the alkynyl group may be appropriately substituted."

[0025] "Alkoxy" refers to a radical of the formula -OR a wherein R a is, as defined above, an alkyl radical having from 1 to 12 carbon atoms (C1-C 12 alkoxy), from 1 to 8 carbon atoms (C1-C8 alkoxy) or from 1 to 6 carbon atoms (C1-C6 alkoxy), or any value within these ranges. Unless otherwise clearly stated herein, the alkoxy group may be appropriately substituted."

[0026] "Aminyl" refers to a radical of the formula -NR a R b wherein R a is H or C1-C6 alkyl and R b is C1-C6 alkyl (as defined above). The C1-C6 alkyl portion of the aminyl group may be appropriately substituted unless otherwise stated."

[0027] "Aminylalkylcycloalkyl" refers to a radical of the formula -R a R b NR c R d wherein R a is cycloalkyl (as defined herein), R b is C1-C6 alkyl (as defined above), R c is H or C1-C6 alkyl,d This refers to a radical that is C1-C6 alkyl. Unless otherwise specified, the cycloalkyl and each C1-C6 alkyl portion of the aminylalkylcycloalkyl group may be substituted as appropriate.

[0028] An "aromatic ring" refers to a cyclic planar molecule or a part of a molecule (i.e., a radical) that has a ring of resonance bonds that exhibits high stability compared to other bond arrangements of the same combination of atoms. Generally, an aromatic ring has a set of atoms covalently bonded on the same plane and contains an even number of π electrons that are not multiples of 4 (i.e., 4n+2 π electrons, where n=0, 1, 2, 3, etc.) (e.g., alternating double and single bonds). Examples of aromatic rings include, but are not limited to, phenyl, naphthenyl, imidazolyl, pyrrolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridonyl, pyridazinyl, and pyrimidonyl. Unless otherwise explicitly stated herein, an "aromatic ring" includes all radicals, which may be substituted as appropriate.

[0029] "Aryl" refers to a group of 6 to 18 carbon atoms (for example, 6 to 10 carbon atoms (C6- 10 This refers to a carbocyclic cyclic radical comprising an aryl group and at least one carbocyclic aromatic ring. In embodiments of the present invention, the aryl radical may be a monocyclic, dicyclic, tricyclic, or tetracyclic cyclic system, and may be a condensed cyclic system or a cross-linked cyclic system. Examples of aryl radicals include, but are not limited to, aryl radicals derived from acetantrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluorantene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Unless otherwise explicitly stated herein, the aryl group may be substituted as appropriate.

[0030] "Cyanoalkyl" refers to an alkyl group containing at least one cyano substituent. The -CN substituent may be located on a primary, secondary, or tertiary carbon. Unless otherwise explicitly stated herein, cyanoalkyl groups may be substituted as appropriate. "Carbocyclic" or "carbocyclic" refers to a cyclic system in which each ring atom is carbon.

[0031] A "cycloalkyl" consists only of carbon atoms and hydrogen atoms, and may be a fused ring system or a bridged ring system, with 3 to 15 ring carbon atoms (C3-C 15 Cycloalkyl), 3 to 10 ring carbon atoms (C3-C 10 This refers to a non-aromatic monocyclic or polycyclic carbocyclic radical having 3 to 8 ring carbon atoms (C3-C8 cycloalkyl), or any value within this range (e.g., 3 to 4 carbon atoms (C3-C4 cycloalkyl)), which is saturated or partially unsaturated and bonded to the rest of the molecule by a single bond. Examples of monocyclic radicals include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of polycyclic radicals include adamantyl, norbornyl, dekalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and similar. Unless otherwise explicitly stated herein, cycloalkyl groups may be substituted as appropriate.

[0032] "Alkylcycloalkyl" is defined by formula -R a R b A radical group of which, in the formula, R a is a cycloalkyl group, and R b is an alkyl group (as defined above), a radical group. Unless otherwise explicitly stated herein, alkylcycloalkyl groups may be substituted as appropriate.

[0033] "Condensed" refers to any ring structure described herein that is condensed with another ring structure.

[0034] "Halo" refers to bromo, chloro, fluoro, or iodine.

[0035] "Haloalkyl" refers to an alkyl radical (as defined above) that is substituted with one or more halo radicals (as defined above), such as trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and the like. Unless otherwise explicitly stated herein, haloalkyls may be substituted as appropriate.

[0036] "Halocycloalkyl" refers to a cycloalkyl radical (as defined above) that is substituted with one or more halo radicals (as defined above), such as trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and the like. Unless otherwise explicitly stated herein, halocycloalkyls may be substituted as appropriate.

[0037] "Haloalkylcycloalkyl" is a compound of the formula -R a R b A radical group of which, in the formula, R a is a cycloalkyl group, and R b is a radical group that is a haloalkyl group (as defined above). Unless otherwise explicitly stated herein, haloalkylcycloalkyl groups may be substituted as appropriate.

[0038] "Halocycloalkylalkyl" is a compound of the formula -R a R b A radical group of which, in the formula, R a R is an alkyl group, b is a radical group that is a halocycloalkyl group (as defined above). Unless otherwise explicitly stated herein, the halocycloalkyl group may be substituted as appropriate.

[0039] "Heterocyclylcycloalkyl" is a compound of the formula -R a R b A radical group of which, in the formula, R a is a cycloalkyl group, and R b This refers to a radical group that is a heterocyclyl group (as defined above). Unless otherwise explicitly stated herein, heterocyclylcycloalkyl groups may be substituted as appropriate.

[0040] "Hydroxyalkyl" refers to an alkyl radical (as defined above) that is substituted with one or more hydroxyl radicals. The hydroxyalkyl radical is bonded to the main chain via an alkyl carbon atom. Unless otherwise explicitly stated herein, the hydroxyalkyl group may be substituted as appropriate.

[0041] "Heterocyclyl" refers to a 3- to 18-membered, e.g., 3- to 10-membered or 3- to 8-membered, non-aromatic ring radical having 1 to 10 ring carbon atoms (e.g., 2 to 10) and 1 to 6 ring heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. Unless otherwise explicitly stated herein, heterocyclyl radicals are partially or completely saturated and may be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, including fused ring systems, spirocyclic ring systems, and / or bridging ring systems. The nitrogen, carbon, and sulfur atoms in the heterocyclyl radical may be oxidized as appropriate, and the nitrogen atom may be quaternized as appropriate. Examples of such heterocyclyl radicals include, but are not limited to, dioxolanil, thienyl[1,3]dithianil, decahydroisoquinolyl, furanolil, imidazolinil, imidazolidinil, isothiazolidinil, isoxazolidinil, morpholinil, octahydroindolyl, octahydroisoindolyl, hexahydro-1H-pyrrolidine, 2-oxopiperazinil, 2-oxopiperidinil, 2-oxopyrrolidinil, oxazolidinil, oxylanil, piperidinil, piperadinil, 4-piperidonil, azetidinil, pyrrolidinil, pyrazolidinil, quinuclidinil, thiazolidinil, tetrahydrofuryl, trithianil, tetrahydropyranil, thiomorpholinil, thiamorpholinil, 1-oxo-thiomorpholinil, and 1,1-dioxo-thiomorpholinil. Unless otherwise explicitly stated herein, heterocyclyl groups may be substituted as appropriate.

[0042] "Haloheterocyclyl" refers to a heterocyclyl group containing at least one halo substituent. The halo substituent may be located on a primary, secondary, or tertiary carbon. Unless otherwise explicitly stated herein, the haloheterocyclyl group may be substituted as appropriate.

[0043] "Haloheterocyclylalkyl" is a compound of formulas -R a R b A radical group of which, in the formula, R a R is an alkyl group, bThis refers to a radical group that is a haloheterocyclyl group (as defined above). Unless otherwise explicitly stated herein, the haloheterocyclylalkyl group may be substituted as appropriate.

[0044] "Heterocyclylalkyl" is a compound of formulas -R a R b A radical group of which, in the formula, R a R is an alkyl group, b This refers to a radical group that is a heterocyclyl group (as defined above). Unless otherwise explicitly stated herein, heterocyclylalkyl groups may be substituted as appropriate.

[0045] "Heteroaryl" refers to a 5- to 18-membered, for example, 5- to 6-membered cyclic radical containing 1 to 6 cyclic heteroatoms selected from the group consisting of 1 to 13 cyclic carbon atoms, nitrogen, oxygen, and sulfur, and at least one aromatic ring. Heteroaryl radicals may be monocyclic, bicyclic, tricyclic, or tetracyclic systems, and may be fused or bridging systems. The nitrogen, carbon, or sulfur atoms in the heteroaryl radical may be oxidized as appropriate, and the nitrogen atom may be quaternized as appropriate. Examples include azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranil, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanil, benzonaphthofuranil, benzoxazolyl, benzodioxolyl, benzodioxynil, benzopyranil, benzopyranonil, benzofuranil, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinil, dibenzofuranil, dibenzothiophenyl, furanil, isothiazolyl, imidazolyl, indazolyl, indazolyl, indazolyl, isoindri Examples include, but are not limited to, indolinyl, isoindolinyl, isoquinolyl, indolidinyl, isoxazolyl, naphthilidinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, 1-oxidepyridinyl, 1-oxidepyrimidinyl, 1-oxidepyradinyl, 1-oxidepyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxadinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyridinyl, pyrimidinyl, pyridadinyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl). Unless otherwise explicitly stated herein, heteroaryl groups may be substituted as appropriate.

[0046] Oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, and 1,3,4-thiadiazolyl have the following structures, respectively: [ka] This refers to oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, and 1,3,4-thiadiazolyl, which are oxazolyl, isoxa It is covalently bonded to one of the carbon atoms in the ring of zolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, and 1,3,4-thiadiazolyl, and to the rest of the molecule.

[0047] As used herein, the term “substituted” means any of the above groups (e.g., alkyl, alkenyl, alkylene, alkylcarbonyl, alkoxy, alkoxyalkyl, aminylalkyl, aryl, cyanoalkyl, cycloalkyl, haloalkyl, heterocyclyl, heterocyclene, heterocyclylalkyl, heteroaryl, heteroarylalkyl, and / or hydroxylalkyl) in which at least one hydrogen atom (e.g., 1, 2, 3, or all hydrogen atoms) is substituted by bonding with a non-hydrogen substituent. Examples of non-hydrogen substituents include, but are not limited to, amino, carboxyl, cyano, hydroxyl, halo, nitro, oxo, thiol, thioxo, alkyl, alkenyl, alkylcarbonyl, alkoxy, aryl, cyanoalkyl, cycloalkyl, haloalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, and / or hydroxylalkyl substituents, each of which may also be appropriately substituted with one or more of the above substituents.

[0048] In some specific embodiments, optional substitutions include halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, C3-C8 halocycloalkyl, and C6-C 10 The group is independently selected from aryls, 5- or 6-membered heteroaryls, C1-C6 alkoxys, and 3- to 8-membered heterocyclines.

[0049] The terms “effective dose” or “therapeutic effective dose” refer to the amount of a compound described herein sufficient to achieve the intended use, such as the treatment of a disease as defined below. The therapeutic effective dose may vary depending on the intended therapeutic use (in vivo) or the subject and condition being treated, e.g., the subject’s weight and age, the severity of the condition, the method of administration, and similar factors, which can be readily determined by those skilled in the art. The term also applies to doses that induce a specific response in target cells, e.g., a reduction in platelet adhesion and / or cell migration. The specific dose will vary depending on the particular selected compound, the administration regimen to be followed, whether it is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system through which the compound is transported.

[0050] As used herein, “treatment” or “to treat” refers to a method for obtaining beneficial or desirable outcomes, such as therapeutic and / or preventive effects, with respect to a disease, disorder, or medical condition, but not limited to these. Therapeutic utility means the eradication or remission of the underlying disease being treated. Therapeutic utility is also achieved by the eradication or remission of one or more physiological symptoms associated with the underlying disease, such that the subject shows improvement despite still suffering from the underlying disease. Preventive effects include delaying or eliminating the onset of a disease or illness, delaying or eliminating the onset of symptoms of a disease or illness, slowing, stopping, or reversing the progression of a disease or illness, or any combination thereof. In some embodiments, for preventive utility, a composition is administered to a subject at risk of developing a particular disease, or to a subject reporting one or more physiological symptoms of a disease, even if the subject has not been diagnosed with the disease.

[0051] As used herein, the terms “co-administration,” “administered in combination,” and grammatically equivalent terms encompass the administration of two or more drugs to an animal (including humans) such that both drugs and / or their metabolites are present simultaneously in the subject. Co-administration includes administration in separate compositions simultaneously, administration in separate compositions at different times, or administration in a composition containing both drugs.

[0052] "Pharmacologically acceptable salts" include both acid and base addition salts.

[0053] A "pharmaceutically acceptable acid addition salt" refers to a salt that retains the biological effects of a free base, is biologically acceptable, or is biologically suitable for administration to a subject. See, for general information, SM Berge, et al., "Pharmaceutical Salts", J. Pharm. Sci., 1977, 66:1-19, and Handbook of Pharmaceutical Salts, Properties, Selection, and Use, Stahl and Wermuth, Eds., Wiley-VCH and VHCA, Zurich, 2002. A preferred pharmaceutically acceptable acid addition salt is one that is pharmacologically effective, suitable for contact with patient tissues, and does not cause excessive toxicity, inflammation, or allergic reactions. Pharmaceutically acceptable acid addition salts include inorganic acids (but not limited to these, e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and similar substances) and organic acids (but not limited to these, e.g., acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, It is formed with glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphate, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucinic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, and similar substances.

[0054] A "pharmaceutically acceptable base addition salt" refers to a salt that retains the biological effects of a free acid, is biologically acceptable, or is biologically suitable for administration to a subject. See, in general, SM Berge, et al., "Pharmaceutical Salts", J. Pharm. Sci., 1977, 66:1-19, and Handbook of Pharmaceutical Salts, Properties, Selection, and Use, Stahl and Wermuth, Eds., Wiley-VCH and VHCA, Zurich, 2002. A preferred pharmaceutically acceptable base addition salt is one that is pharmacologically effective, suitable for contact with patient tissues, and does not cause excessive toxicity, inflammation, or allergic reactions. pharmaceutically acceptable base addition salts are prepared by adding an inorganic or organic base to a free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and similar salts. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, primary, secondary, and tertiary amines, substituted amines such as naturally occurring substituted amines, cyclic amines, and salts of basic ion exchange resins (e.g., ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydravamin, choline, betaine, benetamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like). Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0055] In some embodiments, pharmaceutically acceptable salts include quaternary ammonium salts such as quaternary amine alkyl halides (e.g., methyl bromide) salts.

[0056] The terms “antagonist” and “inhibitor” are used interchangeably and refer to compounds that have the activity to inhibit the biological function of a target protein by inhibiting the activity or expression of the protein (e.g., the association of the NLRP3 inflammasome or NEK7 or NLRP3 inflammasome-NEK7). Thus, the terms “antagonist” and “inhibitor” are defined in the context of the biological role of the target protein. Preferred antagonists as used herein are those that specifically interact with (e.g., bind to) the target, but compounds that inhibit the biological activity of the target protein by interacting with other members of the signaling pathway to which the target protein is a member are also clearly included within the scope of this definition. Preferred biological activities inhibited by antagonists are those related to tumor development, growth, or spread.

[0057] As used herein, the term “agonist” refers to a compound that has the ability to induce or enhance the biological function of a target protein by inhibiting the activity or expression of that target protein. Therefore, the term “agonist” is defined in the context of the biological role of the target polypeptide. Preferred agonists as used herein are those that specifically interact with (e.g., bind to) the target, but compounds that induce or enhance the biological activity of the target polypeptide by interacting with other members of the signaling pathway to which the target polypeptide is a member are also clearly included within this definition.

[0058] "Signal transduction" is the process by which stimuli or inhibitory signals are transmitted to and within cells, triggering intracellular responses.

[0059] The term "selective inhibition" or "selectively inhibiting" refers to the ability of a biological agent to preferentially reduce target signaling activity compared to off-target signaling activity, either directly or indirectly, through interaction with the target.

[0060] "Subject" refers to an animal such as a mammal, for example, a human. The methods described herein may be useful in both therapeutic and veterinary applications for humans. In some embodiments, the subject is a mammal, and in some embodiments, the subject is a human.

[0061] "Mammals" include both humans and domesticated animals (such as laboratory animals and pets (e.g., cats, dogs, pigs, cows, sheep, goats, horses, rabbits)) and non-domesticated animals (such as wild animals and their relatives).

[0062] The term "prodrug" is intended to refer to a compound that may be converted under physiological conditions or by solvolysis to a bioactive compound described herein (e.g., the compound of structure (I)). Therefore, the term "prodrug" refers to a precursor of a pharmaceutically acceptable bioactive compound. In some embodiments, a prodrug is inactive when administered to a subject but is converted in vivo to an active compound, for example, by hydrolysis. Prodrug compounds often provide advantages in mammals such as solubility, histocompatibility, or delayed release (see, e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam)). Discussions of prodrugs are found in “Pro-drugs as Novel Delivery Systems,” ACS Symposium Series, Vol. 14 and Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are fully incorporated herein by reference. The term “prodrug” is also intended to include any covalent carrier that releases the active compound in vivo when such prodrug is administered to a mammalian subject. Prodrugs of active compounds described herein are generally prepared by modifying a functional group in the active compound in such a way that the modification is cleaved either by conventional procedures or in vivo to become the parent active compound. Prodrugs include compounds in which a hydroxyl, amino, or thiol group is bonded to any group that, when the prodrug of the active compound is administered to a mammalian subject, cleaves to form a free hydroxyl, free amino, or free mercapto group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of the hydroxyl functional group in the active compound, or acetamide, formamide, and benzamide derivatives of the amine functional group.

[0063] The term "in vivo" refers to events that occur within the subject's body.

[0064] The embodiments disclosed herein are also intended to encompass all pharmaceutically acceptable compounds of structure (I).

[0065] Certain embodiments are also intended to encompass in vivo metabolites of the disclosed compounds. Such products may arise primarily from enzymatic processes, such as oxidation, reduction, hydrolysis, amidation, or esterification of the administered compound. Therefore, embodiments include compounds produced by processes involving the administration of the disclosed compounds to mammals for a period sufficient to yield their metabolites. Such products are generally identified by administering a radiolabeled compound of the disclosed to an animal (such as a rat, mouse, guinea pig, or monkey) or a human in a detectable dose, allowing sufficient time for metabolism to occur, and then isolating the conversion product from urine, blood, or other biological samples.

[0066] The terms "stable compound" and "stable structure" are intended to indicate compounds that are robust enough to survive isolation from the reaction mixture to a usable purity and formulation into an effective therapeutic agent.

[0067] Often, crystallization results in solvates of the compounds disclosed herein. As used herein, the term “solvate” refers to an aggregate comprising one or more compounds of the Disclosure together with one or more solvent molecules. In some embodiments, the solvent is water, in which case the solvate is a hydrate. Or, in other embodiments, the solvent is an organic solvent. Thus, the compounds of the Disclosure may exist as hydrates (such as monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, and the like) and their corresponding solvated forms. In some embodiments, the compounds of the Disclosure are true solvates, while in others, the compounds of the Disclosure are merely holding added water or a mixture of water and some added solvent.

[0068] "Optional" or "as appropriate" means that the event or situation described thereafter may or may not occur, and that the description includes both cases in which such event or situation occurs and cases in which it does not. For example, "aryls that may be substituted as appropriate" means that the aryl radical may or may not be substituted, and that the description includes both substituted aryl radicals and aryl radicals without substituents.

[0069] "Pharmaceutical composition" refers to a formulation of a medium generally accepted in the art for the delivery of the compounds of this disclosure to a mammal, such as a human. Such a medium may include all pharmaceutically acceptable carriers, diluents, or additives for that purpose.

[0070] "Pharmacologically acceptable carriers, diluents or additives" include, but are not limited to, any adjuvants, carriers, additives, flow enhancers, sweeteners, diluents, preservatives, colorants, flavor enhancers, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers.

[0071] A “stereoisomer” refers to a compound that consists of the same atoms bonded together by the same bonds but has a different three-dimensional structure and is not interchangeable. This disclosure intends various stereoisomers and mixtures thereof and includes “enantiomers,” which refer to two stereoisomers that are mirror images of each other and whose molecules cannot be superimposed.

[0072] The compounds of this disclosure (i.e., compounds of structure (I)) or their pharmaceutically acceptable salts may contain one or more geometrically asymmetric centers, which may give rise to stereoisomers (e.g., enantiomers, diastereomers, and other stereoisomers defined, for example, as (R)- or (S)- from an absolute stereochemistry standpoint, or as (D)- or (L)- for amino acids). Embodiments therefore include all such conceivable isomers, as well as their racemic and optically pure forms. The optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or they may be resolved using conventional techniques, e.g., chromatography and fractional crystallization. Conventional techniques for preparing / isolating each enantiomer include chiral synthesis from a suitable optically pure precursor, or resolution of racemic mixtures (or racemic mixtures of salts or derivatives) using, for example, chiral high-pressure liquid chromatography (HPLC). Where a compound described herein contains an olefin double bond or other geometrically asymmetrical center, unless otherwise specified, the compound is intended to include both E and Z geometric isomers. Similarly, all tautomers are also intended to be included.

[0073] Embodiments of the present disclosure include rotational isomers and conformationally restricted states of all kinds of compounds of the present invention. Atropisomers, which are stereoisomers arising from binding rotations of single bonds, are also included (where steric strain or other inducing energy differences create a barrier to rotation that is high enough to allow isolation of each conformational isomer). For example, certain compounds of the present disclosure may exist as a mixture of atropisomers, and the presence of one atropisomer may be purified or concentrated.

[0074] In some embodiments, the compound of structure (I) is a mixture of enantiomers or diastereomers. In other embodiments, the compound of structure (I) is substantially a single enantiomer or diastereomer.

[0075] "Tautomerism" refers to the transfer of a proton from one atom in a molecule to another atom in the same molecule. Embodiments therefore include tautomers of the disclosed compounds.

[0076] The chemical nomenclature and chemical diagrams used herein are a modified form of IUPAC nomenclature, using the ACD / Name Version 9.07 software program and / or the ChemDraw Professional Version 17.0.0.206 software naming program (CambridgeSoft). For complex chemical names used herein, substituents are generally named before the group to which they are bonded. For example, cyclopropylethyl contains an ethyl skeleton with a cyclopropyl substituent. Except as described below, all bonds are shown in the chemical diagrams herein, except for all bonds on some carbon atoms (assuming there are enough hydrogen atoms to satisfy the valence).

[0077] compound This disclosure provides compounds (including pharmaceutically acceptable salts, stereoisomers, and prodrugs) that can modulate the activity of the NLRP3 inflammasome.

[0078] Embodiments of this disclosure have the following structure (I): [ka] [In the formula, A is C6-C 10 Ariel, C3-C 10 Cycloalkyl, 3-10 membered heterocyclyl, or 5-6 membered monocyclic heteroaryl, each having one or more R 5 This may be substituted as appropriate; X is either N or CH; Y is either CHOH or NH; R 1 is H or C1-C6 alkyl; R 2These are C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3- to 8-membered heterocyclyl, or 5 or 6-membered heteroaryl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyl; R 3 The heteroaryl compounds are selected from oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl and 1,3,4-thiadiazolyl, each being an amino, halo, cyano, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl They may be appropriately substituted with one or more substituents selected from chloroalkyl, C3-C8 alkylcycloalkyl, C3-C8 haloalkylcycloalkyl, C3-C8 aminylalkylcycloalkyl, C1-C6 cyanoalkyl, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered heterocyclylcycloalkyl, 3- to 8-membered haloheterocyclyl, 3- to 8-membered haloheterocyclylalkyl, C3-C8 halocycloalkyl and C3-C8 halocycloalkylalkyl, and combinations thereof; R 4 These include H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3-8 member heterocyclyl, and C6-C 10 An aryl or a 5- or 6-membered heteroaryl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, and C1-C6 alkoxy; R 5These are, independently, halo, cyano, C1-C6 alkyl, C1-C6 hydroxylalkyl, or C1-C6 haloalkyl. The present invention provides compounds having the property, or pharmaceutically acceptable salts, stereoisomers, or prodrugs thereof.

[0079] In some embodiments of structure (I), A is C6-C 10 Ariel, C3-C 10 Cycloalkyl, 3-10 membered heterocyclyl, or 5-6 membered monocyclic heteroaryl, each having one or more R 5 This may be substituted as appropriate; X is either N or CH; Y is either CHOH or NH; R 1 is H or C1-C6 alkyl; R 2 These are C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3- to 8-membered heterocyclyl, or 5 or 6-membered heteroaryl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyl; R 3 is a heteroaryl selected from oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl and 1,3,4-thiadiazolyl, each of which may be appropriately substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl and C3-C8 halocycloalkyl; R 4These include H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3-8 member heterocyclyl, and C6-C 10 An aryl or a 5- or 6-membered heteroaryl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, and C1-C6 alkoxy; R 5 These are, independently, halo, C1-C6 alkyl, C1-C6 alkyl, or C1-C6 haloalkyl.

[0080] One embodiment is a compound of structure (I): [In the formula, A is C6-C 10 Ariel, C3-C 10 Cycloalkyl, 3-10 membered heterocyclyl, or 5-6 membered monocyclic heteroaryl, each having one or more R 5 This may be substituted as appropriate; X is either N or CH; Y is either CHOH or NH; R 1 is H or C1-C6 alkyl; R 2 is H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3- to 8-membered heterocyclyl, or 5 or 6-membered heteroaryl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyl; R 3The heteroaryl compounds are selected from oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl and 1,3,4-thiadiazolyl, each being an amino, halo, cyano, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl They may be appropriately substituted with one or more substituents selected from chloroalkyl, C3-C8 alkylcycloalkyl, C3-C8 haloalkylcycloalkyl, C3-C8 aminylalkylcycloalkyl, C1-C6 cyanoalkyl, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered heterocyclylcycloalkyl, 3- to 8-membered haloheterocyclyl, 3- to 8-membered haloheterocyclylalkyl, C3-C8 halocycloalkyl and C3-C8 halocycloalkylalkyl, and combinations thereof; R 4 These include H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3-8 member heterocyclyl, and C6-C 10 An aryl or a 5- or 6-membered heteroaryl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, and C1-C6 alkoxy; R 5 Each of these is independently a halo, cyano, C1-C6 alkyl, C1-C6 hydroxylalkyl, C1-C6 alkoxy, or C1-C6 haloalkyl. The present invention provides a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof.

[0081] In one embodiment, R 1 In another embodiment, R 1These are C1-C6 alkyl groups, such as methyl groups.

[0082] In one embodiment, a compound of structure (I) (wherein R 2 The provided molecules are branched C4-C6 alkyl, C3-C4 cycloalkyl, C3-C8 heterocyclyl, or 5- or 6-membered heteroaryl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyl.

[0083] In another embodiment, a compound of structure (I) (wherein R 2 The provided molecules are branched C4-C6 alkyl, C3-C4 cycloalkyl, or C3-C8 heterocyclyl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyl.

[0084] In a particular embodiment, R 2 The substituents are cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyls.

[0085] In different embodiments, R 2 The substituents are methyl, isopropyl, 2-methylpropyl, or allyl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyls.

[0086] In different embodiments, R2 The substituents are methyl, ethyl, isopropyl, 2-methylpropyl, or allyl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyls.

[0087] In other embodiments, R 2 The substituents are oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, or dioxidetetrahydrothiophenyl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyls.

[0088] In other embodiments, R 2 The substituents are oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, azetidinyl, or dioxidetetrahydrothiophenyl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyls.

[0089] In other embodiments, R 2 is oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, azetidinyl, pyrrolidinyl, or dioxidetetrahydrothiophenyl, each of which may be appropriately substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyl.

[0090] In a further embodiment, R 2is pyridinyl, optionally substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyl.

[0091] In any of the foregoing embodiments, R 2 is unsubstituted. In other of the foregoing embodiments, R 2 is substituted with one or more of hydroxyl and fluoro.

[0092] In any of the foregoing embodiments, R 2 is unsubstituted. In other of the foregoing embodiments, R 2 is substituted with one or more of hydroxyl, methyl, methoxy, and fluoro.

[0093] In a more particular embodiment, R 2 has one of the following structures:

Chemical formula

[0094] In a further particular embodiment, R 2 has one of the following structures:

Chemical formula

[0095] In a further particular embodiment, R 2 has one of the following structures:

Chemical formula

[0096] In some embodiments, R 2It may be optionally substituted with one or more substituents selected from the group consisting of halo, hydroxyl, cyano, aminyl, C1-C6 alkyl, and 3- to 8-membered heterocyclyl.

[0097] In some embodiments, R 2 has the following structure:

Chemical formula

[0098] In any of the above-described embodiments, R 3 is oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl or 1,3,4-oxadiazolyl, each of which may be optionally substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl and C3-C8 halocycloalkyl. For example, in one embodiment, R 3 is isoxazolyl and may be optionally substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl and C3-C8 halocycloalkyl. In a further specific embodiment, R 3 is substituted with C1-C6 alkyl, C1-C6 haloalkyl, C3-C8 cycloalkyl or C3-C8 halocycloalkyl.

[0099] In a further embodiment, R 3is oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,3,4-oxadiazolyl, thiazolyl, isothiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, or 1,2,4-triazolyl, each of which may be appropriately substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, cyano, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl and C3-C8 halocycloalkyl, or combinations thereof.

[0100] In one embodiment, R 3 isoxazolyl, which may be appropriately substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, cyano, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl and C3-C8 halocycloalkyl, or combinations thereof.

[0101] In one embodiment, R 3 This is thiazolyl and may be appropriately substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, cyano, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl and C3-C8 halocycloalkyl, or combinations thereof.

[0102] In one embodiment, R 3 isothiazolyl, which may be appropriately substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, cyano, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl and C3-C8 halocycloalkyl, or combinations thereof.

[0103] In one embodiment, R 3 is 1,2,4-thiadiazolyl, which may be appropriately substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, cyano, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl and C3-C8 halocycloalkyl, or combinations thereof.

[0104] In one embodiment, R 3 is 1,3,4-thiadiazolyl, which may be appropriately substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, cyano, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl and C3-C8 halocycloalkyl, or combinations thereof.

[0105] In one embodiment, R 3 is 1,3,4-oxadiazolyl, which may be appropriately substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, cyano, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl and C3-C8 halocycloalkyl, or combinations thereof.

[0106] In one embodiment, R 3 is 1,2,4-triazolyl and may be appropriately substituted with one or more substituents selected from halo, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, cyano, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl and C3-C8 halocycloalkyl, or combinations thereof.

[0107] In a further embodiment, R 3 This is substituted with C1-C6 alkyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, cyano, C1-C6 aminyl, C1-C6 hydroxylalkyl, 3- to 8-membered heterocyclyl or C3-C8 halocycloalkyl, or a combination thereof.

[0108] In various embodiments, R 3 The structure is as follows: [ka] It has one of the following.

[0109] In a further embodiment, R 3 The structure is as follows: [ka] It has one of the following.

[0110] In a further embodiment, R 3 The structure is as follows: [ka] It has one of the following.

[0111] In other embodiments, R 4 In another embodiment, R 4 These are C1-C6 alkyl groups, such as methyl groups.

[0112] In one embodiment, Y is CHOH. In another embodiment, Y is NH.

[0113] In other embodiments, X is N. In yet another embodiment, X is CH.

[0114] In various embodiments, A is C6-C 10 Ariel, C3-C 10A cycloalkyl or a 5-6 member monocyclic heteroaryl, each containing one or more R 6 It may be substituted as appropriate. A is understood to be a divalent radical.

[0115] In one embodiment, A is divalent, and C6-C 10 It may be optionally substituted with aryl atoms. In one embodiment, A may be optionally substituted with a divalent, 3- to 8-membered saturated or partially unsaturated carbocyclic ring. In one embodiment, A may be optionally substituted with a divalent, 3- to 10-membered heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In one embodiment, A may be optionally substituted with a divalent, 5- to 6-membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0116] In one embodiment, A is a divalent group selected from phenyl, pyridinyl, cyclohexyl, and cyclohexenyl, each of which may be substituted as appropriate.

[0117] In other embodiments, A is phenyl. In different embodiments, A is saturated or unsaturated cyclohexyl. In further embodiments, A is pyridinyl.

[0118] In further embodiments, A is pyrimidinyl and may be substituted as appropriate.

[0119] In any of the embodiments described above, A is non-substitutable. In a different embodiment described above, A is one or more R 5 It is replaced by R. For example, in some embodiments, 5 is a halo. In other embodiments, R 5 is fluoro. In other different embodiments, R 5 That is Chlorophyll.

[0120] In some embodiments, R 5is cyano. In some embodiments, R 5 is a C1-C6 alkyl group. In one embodiment, R 5 is methyl. In some embodiments, R 5 is a C1-C6 haloalkyl. In one embodiment, R 5 R is difluoromethyl. In further embodiments, R 5 is a C1-C6 hydroxylalkyl group. In one embodiment, R 5 It is -CH2OH.

[0121] In one embodiment, A is phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctanyl, [4.3.0]bicyclononanyl, [4.4.0]bicyclodecanyl, [2.2.2]bicyclooctanyl, fluorenyl, indanyl, tetrahydronaphthyl, acridinyl, azocinyl, benzimidazolyl, benzofuranil, benzothiofuranil, benzothiophenyl , benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzoisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, carborinyl, chromanil, chromenil, cinnolinil, decahydroquinolinil, dithiadinyl, tetrahydrofuranil, furanil, flazanil, imidazolidinyl, imidazolinil, imidazolyl, 1H-indazolyl, indolenyl, indolinil, indolidine, indolyl, 3-indolyl, Isoindolenyl, isoindolenyl, isobenzofuranil, isochromanil, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl, naphthilidinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthryl Dinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperadinyl, piperidinyl, pteridinyl, prinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridoxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2-pyrrolyl, pyrrrolyl, quinazolinyl, quinolinyl, 4H-quinolidinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiadiadinyl, 1,2,A divalent group selected from 3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, and xanthenyl, each of which may be substituted as appropriate.

[0122] In a particular embodiment, A has the following structure: [ka] It has one of the following.

[0123] In other specific embodiments, A has the following structure: [ka] It has one of the following.

[0124] In one embodiment, the compound of structure (I) is a modulator of the NLRP3 inflammasome.

[0125] In certain embodiments, the compound of structure (I) is an inhibitor of NEK7 in a patient or in a biological sample.

[0126] In various different embodiments, the compound has one of the structures listed in Table 1 below, or is a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof. The compounds in Table 1 are prepared as described in the examples or by methods known in the art, and analyzed by mass spectrometry and / or 1 The results were analyzed by 1H NMR. [Table 1] [Table 2] Table 3 Table 4 Table 5 Table 6 Table 7 Table 8 Table 9 Table 10 Table 11 Table 12 Table 13 Table 14 Table 15 Table 16 Table 17 Table 18 Table 19 Table 20 [Table 21] [Table 22] [Table 23]

[0127] In this explanation, it is understood that combinations of substituents and / or variables in the given formulas are acceptable only if such contributions result in a stable compound.

[0128] In further embodiments, various compounds of the Disclosure, existing in the form of free bases or acids, can be converted to their pharmaceutically acceptable salts by treatment with suitable inorganic or organic bases or acids in a manner known to those skilled in the art. Salts of the compounds of the Disclosure can be converted to the form of free bases or acids by standard techniques.

[0129] The methods for preparing the compounds described herein are shown below. Generally, starting components may be obtained from sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, or synthesized according to information known to those skilled in the art (see, for example, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th edition (Wiley, December 2000)), or prepared as described herein.

[0130] The following general reaction scheme is structure (I): [ka] [In the formula, A, X, Y, R 1 , R 2 , R 3 and R 4 Each of these is as defined herein. This document describes typical preparation methods for compounds or their pharmaceutically acceptable salts, stereoisomers, or prodrugs.

[0131] General reaction scheme 1 The following is a general reaction scheme (where X is in the formula) 1 It is a halogen, and X, R 1 , R 2 An example of a method for preparing amine intermediate B (where A has the meaning described herein) is illustrated. [ka]

[0132] As shown in the general reaction scheme 1, in the presence of a base, functionalizing pyrazolopyrimidine with an alkyl or aryl boronic acid or a suitable electrophile gives intermediate A, which is then arylated with a palladium catalyst (followed by a reduction step if necessary) to give amine intermediate B.

[0133] General reaction scheme 2 The following general reaction scheme illustrates an example of a method for preparing carbamate intermediate C.

Chemical formula

[0134] As shown in the general reaction scheme 2, intermediate C can be prepared by reacting phenyl chloroformate with the indicated heteroaryl amine (amine substitution analog of R 3 ) in the presence of a base. The general reaction scheme 2 shows the preparation of a compound where R 4 is H, but compounds where R 4 is other than H can be prepared by introducing R 4 after the preparation of intermediate I or by a similar method using a properly substituted heteroaryl amine.

[0135] General reaction scheme 3 The following general reaction scheme illustrates an example of a method for preparing a compound of structure (I).

Chemical formula

[0136] Intermediate B and intermediate C are treated with a base (e.g., trimethylamine, etc.) in THF to obtain a compound of structure (I).

[0137] General reaction scheme 4 The following general reaction scheme illustrates an example of a method for preparing a compound of structure (I).

Chemical formula

[0138] Intermediate B is reacted with phenyl chloroformate under appropriate conditions to obtain intermediate C. Next, intermediate C is coupled with an amine in THF using a suitable base (e.g., trimethylamine, DIPEA, DMAP, and similar) to obtain the compound of structure (I).

[0139] Any of the above reaction schemes can be modified at any step to add and / or change substituents or change the order of steps, as needed, at any stage of the overall synthesis of the compound of interest. For example, those skilled in the art may instead prepare a carbamate analog of intermediate B, and R 3 It will be easy to understand that the compound of structure (I) can be prepared by reacting it with an amine analog.

[0140] Those skilled in the art will recognize that in the process of preparing the compounds described herein, the functional groups of intermediate compounds may need to be protected with appropriate protecting groups. Such functional groups include, but are not limited to, hydroxyl, amino, mercapto, and carboxylic acids. Suitable hydroxyl protecting groups include trialkylsilyl or diarylalkylsilyl (e.g., t-butyldimethylsilyl, t-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable amino, amidino, and guanidino protecting groups include t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable mercapto protecting groups include -C(O)-R” (where R is alkyl, aryl, or arylalkyl), p-methoxybenzyl, trityl, and the like. Suitable carboxylic acid protecting groups include alkyl, aryl, or arylalkyl esters. Protecting groups are known to those skilled in the art and can be added or removed as appropriate according to the usual art as described herein. The use of protecting groups is described in detail in Green, TW and PGM Wutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley. As those skilled in the art will recognize, the protecting group may also be a polymer resin (such as Wang resin, Rink resin, or 2-chlorotrityl chloride resin).

[0141] It will be recognized by those skilled in the art that such protected derivatives of the compounds of the present disclosure may not have pharmacological activity on their own, but may be administered to mammals and subsequently metabolized in the body to become pharmacologically active compounds of the present disclosure. Such derivatives are therefore sometimes referred to as “prodrugs.” Prodrugs of the compounds of the present disclosure are included within the scope of embodiments of the present invention.

[0142] Pharmaceutical composition Other embodiments relate to pharmaceutical compositions. A pharmaceutical composition comprises one (or more) of the aforementioned compounds and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is formulated for oral administration. In other embodiments, the pharmaceutical composition is formulated for injection. In further embodiments, the pharmaceutical composition comprises the compounds disclosed herein and additional therapeutic agents (e.g., anticancer agents). Examples of such therapeutic agents, not limited to those described herein, are described below.

[0143] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ophthalmic, transpulmonary, transmucosal, transdermal, vaginal, otorhinolaryngeal, nasal, and topical administration. Furthermore, as just a few examples, parenteral delivery includes intramuscular, subcutaneous, intravenous, intrathecal injection, as well as intrathecal, direct intraventricular, intraperitoneal, lymphatic, and intranasal injection.

[0144] In some embodiments, the compounds described herein are often administered locally (e.g., by direct injection into an organ) rather than systemically, often in depot or sustained-release formulations. In certain embodiments, long-acting formulations are administered by implantation (e.g., subcutaneous or intramuscular) or intramuscular injection. Furthermore, in other embodiments, the compounds are delivered by a targeted drug delivery system, for example, by liposomes coated with organ-specific antibodies. In such embodiments, the liposomes target the organ and are selectively taken up by the organ. In yet another embodiment, the compounds described herein are provided in the form of immediate-release formulations, sustained-release formulations, or intermediate-release formulations. In yet another embodiment, the compounds described herein are administered locally.

[0145] In the therapeutic methods described in embodiments of the present invention, an effective amount of a compound of at least one structure (I) is administered to a subject suffering from or diagnosed with such disease, disorder, or condition. The effective amount or effective dose may be determined by methods such as modeling, dose escalation studies, or clinical trials, for example, by the method or route of administration or drug delivery, the pharmacokinetics of the drug, the severity and course of the disease, disorder, or illness, the subject's previous or ongoing treatment, the subject's health status and response to the drug, and the judgment of the treating physician.

[0146] The compounds described herein are effective over a wide dose range. For example, doses of 10 to 5000 mg, 100 to 5000 mg, 1000 to 4000 mg, and 1000 to 3000 mg per day in the treatment of adults are examples of doses used in some embodiments. The exact dose will depend on the route of administration, the form in which the compound is administered, the patient being treated, the patient's weight, and the choice and experience of the attending physician.

[0147] In some embodiments, the compounds of the present disclosure are administered in single doses. Generally, such administration will be by injection (e.g., intravenous injection) to rapidly introduce the drug. However, other routes may be used as needed. Single doses of the compounds of the present disclosure may also be used for the treatment of acute diseases.

[0148] In some embodiments, the compounds of the Disclosure are administered in multiple doses. In some embodiments, the administration is about once, twice, three, four, five, six times, or six or more times per day. In other embodiments, the administration is about once a month, once every two weeks, once a week, or once every other day. In yet another embodiment, the compounds of the Disclosure and another agent (e.g., an anticancer agent) are administered together about once to about six times per day. In yet another embodiment, the administration of the compounds and agents of the Disclosure continues for less than about seven days. In yet another embodiment, the administration continues for about six, ten, fourteen, or twenty-eight days, two months, six months, or one year or more. In some examples, continuous administration is achieved and maintained as long as necessary.

[0149] Administration of the compounds of this disclosure may be continued for as long as necessary. In some embodiments, the compounds of this disclosure are administered for 1, 2, 3, 4, 5, 6, 7, 14, or 28 days or longer. In some embodiments, the compounds of this disclosure are administered for 28, 14, 7, 6, 5, 4, 3, 2, or less than 1 day. In some embodiments, the compounds of this disclosure are administered continuously for a long period of time, for example, to treat chronic effects.

[0150] In some embodiments, the compounds of this disclosure are administered in individual dosage forms. Due to intersubjective variability in the pharmacokinetics of the compounds, it is known in the art that individualization of the dosing regimen is necessary for optimal therapy.

[0151] In some embodiments, the compounds described herein are formulated into pharmaceutical compositions. In certain embodiments, the pharmaceutical compositions are formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and adjuvants that facilitate the processing of the disclosed compounds into pharmaceutically usable formulations. The appropriate formulation depends on the selected route of administration. Any pharmaceutically acceptable techniques, carriers, and excipients are used as suitable for the formulation of the pharmaceutical compositions described herein. Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999)

[0152] Pharmaceutical compositions comprising one or more compounds of structure (I) and a pharmaceutically acceptable carrier are provided herein.

[0153] There is provided herein a pharmaceutical composition comprising one or more compounds selected from the compounds of structure (I) and a pharmaceutically acceptable diluent, additive, and carrier. In certain embodiments, the described compounds are administered as a pharmaceutical composition in which one or more compounds selected from the compounds of structure (I) are mixed with other active ingredients, such as in combination therapy. All combinations of the active ingredients described in the following combination therapy section and throughout this disclosure are included herein. In certain embodiments, the pharmaceutical composition comprises one or more compounds of structure (I).

[0154] In certain embodiments, the pharmaceutical composition of the compound of structure (I) is a modulator of the NLRP3 inflammasome.

[0155] In certain embodiments, the pharmaceutical composition of the compound of structure (I) inhibits NEK7 when administered to a patient or a biological sample.

[0156] As used herein, a pharmaceutical composition refers to a mixture of one or more compounds selected from the compounds of structure (I) and other chemical components (such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, and / or additives, etc.). In certain embodiments, the pharmaceutical composition facilitates the administration of the compound to a living being. In some embodiments, a therapeutically effective amount of one or more compounds selected from the compounds of structure (I) provided herein is administered to a mammal having a disease, disorder, or condition to be treated in a pharmaceutical composition. In certain embodiments, the mammal is a human. In certain embodiments, the therapeutically effective amount varies depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. The compounds described herein are used alone or in combination with one or more therapeutic agents as components of a mixture.

[0157] In one embodiment, one or more compounds selected from the compounds of structure (I) are formulated in an aqueous solution. In a particular embodiment, the aqueous solution is selected, as just one example, from a physiologically suitable buffer, such as Hanks' solution, Ringer's solution, or saline buffer. In another embodiment, one or more compounds selected from the compounds of structure (I) are formulated for transmucosal administration. In a particular embodiment, the transmucosal formulation includes a penetrating agent suitable for the barrier to be penetrated. In yet another embodiment in which the compounds described herein are formulated for other parenteral injections, suitable formulations include aqueous or non-aqueous solutions. In a particular embodiment, such solution includes a physiologically suitable buffer and / or additives.

[0158] In another embodiment, the compounds described herein are formulated for oral administration. The compounds described herein are formulated by combining the active compound with, for example, a pharmaceutically acceptable carrier or additive. In various embodiments, the compounds described herein are formulated in oral dosage forms, including, but not limited to, tablets, powders, pills, sugar-coated tablets, capsules, liquids, gels, syrups, elixirs, slurries, suspensions, and the like.

[0159] In one embodiment, an oral pharmaceutical is obtained by mixing one or more solid additives with one or more compounds described herein, grinding the resulting mixture as appropriate, adding suitable adjuvants as necessary to obtain a tablet or sugar-coated tablet core, and then processing the granular mixture. Suitable additives include fillers in particular (e.g., sugars such as lactose, sucrose, mannitol, or sorbitol), cellulose preparations (e.g., corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, microcrystalline cellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, etc.), or others (e.g., polyvinylpyrrolidone (PVP or povidone) or calcium phosphate, etc.). In certain embodiments, disintegrants are added as appropriate. Disintegrants include, but are not limited to, cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or its salts (e.g., sodium alginate, etc.).

[0160] In one embodiment, a dosage form (e.g., a sugar-coated tablet core and a tablet) is provided with one or more suitable coatings. In a particular embodiment, a concentrated sugar solution is used to coat the dosage form. The sugar solution optionally comprises additional components (just as an example, gum arabic, talc, polyvinylpyrrolidone, Carbopol gel, polyethylene glycol, and / or titanium dioxide), a lacquer solution, and a suitable organic solvent or solvent mixture. Dyes and / or pigments are also optionally added to the coating for identification. Furthermore, dyes and / or pigments are optionally used to characterize different combinations of doses of the active compound.

[0161] In one embodiment, a therapeutically effective amount of at least one of the compounds described herein is formulated into other oral dosage forms. These oral dosage forms include push-fit capsules made of gelatin, and sealed soft capsules made of gelatin and a plasticizer (e.g., glycerol or sorbitol). In certain embodiments, the push-fit capsule contains the active ingredient in a mixture with one or more fillers. Fillers include, but are not limited to, lactose, binders (e.g., starch), and / or lubricants (e.g., talc or magnesium stearate), and optionally, stabilizers. In other embodiments, the soft capsule contains one or more active compounds dissolved or suspended in a suitable liquid. Suitable liquids include, but are not limited to, one or more fatty oils, liquid paraffin, or liquid polyethylene glycol. Further stabilizers may be added as appropriate.

[0162] In further embodiments, the compounds described herein are formulated for parenteral injection (such as formulations suitable for bolus injection or continuous infusion). In certain embodiments, the injectable formulation is provided in single-dose dosage forms (e.g., in ampoules) or in multi-dose containers. Preservatives are added to the injectable formulation as appropriate. In further embodiments, the pharmaceutical composition is formulated in a form suitable for parenteral injection as a sterile suspension, solution, or emulsion in an oily or aqueous vehicle. The parenteral injection formulation comprises formulation agents as appropriate (e.g., suspending agents, stabilizers, and / or dispersants). In certain embodiments, the pharmaceutical formulation for parenteral administration includes aqueous solutions of the water-soluble active compound. In further embodiments, a suspension of one or more compounds selected from the compounds of structure (I) is prepared as a suitable oily suspension for injection. Suitable lipophilic solvents or vehicles used in the pharmaceutical compositions described herein include, but are not limited to, fatty oils (e.g., sesame oil), synthetic fatty acid esters (e.g., ethyl oleate or triglycerides), or liposomes. In certain embodiments, the aqueous suspension for injection contains a substance that increases the viscosity of the suspension (e.g., sodium carboxymethylcellulose, sorbitol, or dextran). Optionally, the suspension contains a suitable stabilizer or agent that increases the solubility of the compound to enable the preparation of high-concentration solutions. Alternatively, in other embodiments, the active ingredient is in powder form for preparation with a suitable vehicle before use, such as sterile pyrogen-removed water.

[0163] A pharmaceutical composition comprises at least one pharmaceutically acceptable carrier, diluent, or additive, and one or more compounds selected from the compounds of structure (I) described herein as the active ingredient. The active ingredient is in the form of a free acid or free base, or a pharmaceutically acceptable salt. Furthermore, the methods and pharmaceutical compositions described herein include the use of N-oxides, crystalline forms (also called polymorphs), and active metabolites of these compounds having the same type of activity. All tautomers of the compounds described herein fall within the range of compounds presented herein. Furthermore, the compounds described herein include both non-solvated forms and solvated forms with pharmaceutically acceptable solvents (e.g., water, ethanol, and the like). The solvated forms of the compounds presented herein are also considered to be disclosed herein. Furthermore, the pharmaceutical composition may optionally include other agents or pharmaceutical preparations, carriers, adjuvants (e.g., preservatives, stabilizers, wetting agents, or emulsifiers), dissolution accelerators, osmotic salts, buffers, and / or other therapeutically useful substances.

[0164] Methods for preparing compositions containing the compounds described herein include formulating the compounds with one or more inert, pharmaceutically acceptable additives or carriers to form solids, semi-solids, or liquids. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets, and suppositories. Liquid compositions include solutions in which the compounds are dissolved, emulsions containing the compounds, or solutions containing liposomes, micelles, or nanoparticles containing the compounds disclosed herein. Semi-solid compositions include, but are not limited to, gels, suspensions, and creams. The forms of pharmaceutical compositions described herein include liquid solutions or suspensions, solid forms suitable for dissolving or suspending in a liquid before use, or emulsions. These compositions may also, as appropriate, contain small amounts of non-toxic auxiliary substances (e.g., wetting agents, emulsifiers, pH buffers, etc.).

[0165] In some embodiments, a pharmaceutical composition comprising one or more compounds selected from the compounds of structure (I) typically takes the form of a liquid in which the drug exists in a solution, a suspension, or both. Generally, when the composition is administered as a suspension, a first portion of the drug exists in a solution, and a second portion of the drug exists in the form of suspended particles in a liquid matrix. In some embodiments, the liquid composition includes a gel formulation. In other embodiments, the liquid composition is water-soluble.

[0166] In one embodiment, the aqueous suspension contains one or more polymers as a suspending agent. Examples of polymers include water-soluble polymers such as cellulose polymers (e.g., hydroxypropyl methylcellulose) and water-insoluble polymers such as cross-linked carboxyl-containing polymers. Specific pharmaceutical compositions described herein include mucosal-adhering polymers, selected from, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylic acid), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate, and dextran.

[0167] The pharmaceutical composition may also include, as appropriate, a solubilizer that helps dissolve one or more compounds selected from the compounds of structure (I). The term “solubilizer” generally includes agents that result in the formation of micelles or true solutions of the drug. Certain acceptable nonionic surfactants, such as polysorbate 80, are useful as solubilizers, as are ophthalmologically acceptable glycols, polyglycols (e.g., polyethylene glycol 400), and glycol ethers.

[0168] Furthermore, the pharmaceutical composition may contain one or more pH adjusters or buffers as appropriate (such as acids like acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid; bases like sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and trishydroxymethylaminomethane; and buffers like citrate / dextrose, sodium bicarbonate, and ammonium chloride). Such acids, bases, and buffers are included in amounts necessary to maintain the pH of the composition, within acceptable limits.

[0169] The composition may also include, as appropriate, one or more salts in amounts necessary to bring the osmolality of the composition into an acceptable range. Such salts include those having sodium, potassium, or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate, or bisulfite anions. Suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.

[0170] Other pharmaceutical compositions may optionally include one or more preservatives that inhibit microbial activity. Suitable preservatives include mercury-containing substances such as merfen and thiomersal, stabilized chlorine dioxide, and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

[0171] The composition may contain one or more surfactants to enhance physical stability or for other purposes. Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils (e.g., polyoxyethylene (60) hydrogenated castor oil), as well as polyoxyethylene alkyl ethers and alkylphenyl ethers (e.g., octoxynol 10, octoxynol 40).

[0172] The composition may include one or more antioxidants to enhance chemical stability, if necessary. Suitable antioxidants include, but are not limited to, ascorbic acid and sodium metabisulfite.

[0173] In one embodiment, the aqueous suspension composition is packaged in single doses in a container that cannot be resealed. Alternatively, a resealable multi-dose container is used, in which case the composition typically contains a preservative.

[0174] In other embodiments, other delivery systems for hydrophobic pharmaceutical compounds are utilized. Liposomes and emulsions are examples of delivery vehicles or carriers useful herein. In some embodiments, organic solvents, such as N-methylpyrrolidone, are also utilized. In further embodiments, the compounds described herein are delivered using sustained-release systems, such as a semipermeable matrix of a solid hydrophobic polymer containing the therapeutic agent. Various sustained-release materials are useful herein. In some embodiments, sustained-release capsules release the compound over a period of several weeks to over 100 days. Depending on the chemical properties and biological stability of the therapeutic reagent, further strategies for protein stabilization are utilized.

[0175] In one embodiment, the formulations described herein include one or more antioxidants, metal chelating agents, thiol-containing compounds, and / or other common stabilizers. Examples of such stabilizers include, but are not limited to, (a) glycerol in approximately 0.5% to 2% w / v, (b) methionine in approximately 0.1% to 1% w / v, (c) monothioglycerol in approximately 0.1% to 2% w / v, (d) EDTA in approximately 1 mM to 10 mM, (e) ascorbic acid in approximately 0.01% to 2% w / v, (f) polysorbate 80 in approximately 0.003% to 0.02% w / v, (g) polysorbate 20 in approximately 0.001% to 0.05% w / v, (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrin, (l) pentosan polysulfate and other heparin analogs, (m) divalent cations such as magnesium and zinc, or (n) combinations thereof.

[0176] In some embodiments, the concentrations of one or more compounds selected from the compounds of structure (I) provided in the pharmaceutical compositions of the present disclosure are 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25%, 19%, 18.75%, 18.50%, 18.25%, 18%, 17.75%, 17.50%, 17.25%, 17%, 16.75%, 16.50%, 16.25%, 16%, 15.75%, 15.50%, 15.25%, 15%, 14.75%, 14.50%, 14.25%, 14%, 13.75%, 13.50%, 13.25%. 13%, 12.75%, 12.50%, 12.25% 12%, 11.75%, 11.50%, 11.25% 11%, 10.75%, 10.50%, 10.25% 10%, 9.75%, 9.50%, 9.25% 9%, 8.75%, 8.50%, 8.25% 8%, 7.75%, 7.50%, 7.25% 7%, 6.75%, 6.50%, 6.25% 6%, 5.75%, 5.50%, 5.25% 5%, 4.75%, 4.50%, 4.25%, 4%, 3.75%, 3.50%, 3.25%, 3%, 2.75%, 2.50%, 2.25%, 2%, 1.75%, 1.50%, 125% , 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w / w, w / v, or v / v or more.

[0177] In some embodiments, the concentration of one or more compounds selected from the compounds of structure (I) provided in the pharmaceutical compositions of the present disclosure is approximately 0.0001% to approximately 50%, approximately 0.001% to approximately 40%, approximately 0.01% to approximately 30%, approximately 0.02% to approximately 29%, approximately 0.03% to approximately 28%, approximately 0.04% to approximately 27%, approximately 0.05% to approximately 26%, approximately 0.06% to approximately 25%, and approximately 0. The ranges are approximately 0.7% to 24%, approximately 0.08% to 23%, approximately 0.09% to 22%, approximately 0.1% to 21%, approximately 0.2% to 20%, approximately 0.3% to 19%, approximately 0.4% to 18%, approximately 0.5% to 17%, approximately 0.6% to 16%, approximately 0.7% to 15%, approximately 0.8% to 14%, approximately 0.9% to 12%, and approximately 1% to 10%, all within the ranges of w / w, w / v, or v / v.

[0178] In some embodiments, the amounts of one or more compounds selected from the compounds of structure (I) provided in the pharmaceutical compositions of the present disclosure are 10g, 9.5g, 9.0g, 8.5g, 8.0g, 7.5g, 7.0g, 6.5g, 6.0g, 5.5g, 5.0g, 4.5g, 4.0g, 3.5g, 3.0g, 2.5g, 2.0g, 1.5g, 1.0g, 0.95g, 0.9g, 0.85g, 0.8g, 0.75g, 0.7g, 0.65g, 0.6g, 0.55g, 0.5g, 0.45g, 0.4g, 0.35g, 0.3g, 0.25g, 0.2g, 0.15g, 0.1g, 0.09g, 0.08g, 0.07g, 0.06g, 0.05g, 0.04g, 0.03g, 0.02g, 0.01g, 0.009g, 0.008g, 0.007g, 0.006g, 0.005g, 0.004g, 0.003g, 0.002g, 0.001g, 0.0009g, 0.0008g, 0.0007g, 0.0006g, 0.0005g, 0.0004g, 0.0003g, 0.0002g, or 0.0001g or less.

[0179] In some embodiments, the amount of one or more compounds selected from the compounds of structure (I) provided in the pharmaceutical compositions of the present disclosure is in the range of 0.0001 to 10 g, 0.0005 to 9 g, 0.001 to 8 g, 0.005 to 7 g, 0.01 to 6 g, 0.05 to 5 g, 0.1 to 4 g, 0.5 to 4 g, or 1 to 3 g.

[0180] Packaging materials used for packaging the pharmaceutical compositions described herein include, for example, those found in U.S. Patent Nos. 5,323,907, 5,052,558, and 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging materials suitable for the selected formulation and the intended method of administration and mode of treatment. For example, a container may contain one or more compounds described herein, either in the composition or in combination with another agent disclosed herein. The container may have a sterile access port as appropriate (for example, the container may be a solution bag for intravenous injection or a vial with a stopper that can be punctured with a subcutaneous needle). Such a kit may contain the compounds together with an identifying description or label or instructions for its use in the manner described herein.

[0181] For example, a kit generally includes one or more additional containers, each containing one or more of the various materials (e.g., reagents and / or devices, which may be concentrated as appropriate) that are desirable from a commercial and user perspective for the use of the compounds described herein. Examples of such materials, though not limited to, include buffers, diluents, filters, needles, syringes, carriers, packaging, containers, vials and / or tube labels, and accompanying documentation with instructions for use. A complete set of instructions would also generally be included. Labels may be on the container or attached to the container, as appropriate. For example, if the letters, numbers, or other characters forming the label are affixed, cast, or etched onto the container itself, the label is on the container; if the label is included, for example, as accompanying documentation, within a container or carrier that also holds the container, the label is attached to the container. Furthermore, labels are used to indicate that the contents are intended for a specific therapeutic application. Additionally, labels indicate how to use the contents, such as the methods described herein. In some embodiments, the pharmaceutical composition is contained in a pack or dispenser device containing one or more single-dose dosage forms of the compounds provided herein. The pack contains, for example, metal foil or plastic foil (such as a blister pack). Alternatively, the pack or dispenser device is accompanied by instructions for administration. Alternatively, the pack or dispenser is accompanied by a notice related to the container in the format prescribed by the administrative agency that regulates the manufacture, use, or sale of pharmaceuticals, the notice reflecting agency approval of the form of the drug for administration to humans or animals. Such notice is, for example, labeling approved as a prescription drug by the U.S. Food and Drug Administration, i.e., an approved product insert. In some embodiments, compositions containing the compounds described herein, formulated in a suitable pharmaceutical carrier, are prepared, placed in a suitable container, and labeled for the treatment of the indicated disease.

[0182] method Embodiments of this disclosure are useful as modulators of the NLRP3 inflammasome via inhibition of NEK7 in host animal species. Therefore, the compound of structure (I) is also useful in the treatment of diseases mediated by effector signaling molecules such as IL-1β and IL-18.

[0183] The host or patient may belong to any mammalian species, such as primates (especially humans), rodents (mice, rats, and hamsters, etc.); rabbits; horses, cattle, dogs, cats, etc. Animal models are of interest for experimental investigations and provide models for the treatment of human diseases.

[0184] In one embodiment, the present disclosure is useful as an inhibitor of the NLRP3 inflammasome activation mechanism. Therefore, the compound of structure (I) is also useful for treating diseases caused by its activation in host animal species.

[0185] In another embodiment, the compound of structure (I) is useful as an inhibitor of the NLRP3 (protein)-NEK7 (protein) interaction. Therefore, the compound is also useful for treating diseases caused by the association of NLRP3-NEK7 in host animal species.

[0186] In one embodiment, the compound of structure (I) is useful for treating human diseases mediated by an effector selected from the group consisting of IL-1β, IL-18, and caspase-1.

[0187] Embodiments of the present invention also relate to the use of the compound of structure (I) and / or physiologically acceptable salts thereof for prophylactic treatment or therapeutic intervention and / or monitoring of diseases caused, mediated, and / or regulated by NLRP3 inflammasome activity. Furthermore, embodiments of the present invention relate to the use of the compound of structure (I) and / or physiologically acceptable salts thereof for the purpose of producing drugs for prophylactic treatment or therapeutic intervention and / or monitoring of diseases caused, mediated, and / or regulated by NLRP3 inflammasome activity. In one embodiment, the present invention provides the use of the compound of structure I or a physiologically acceptable salt thereof for the purpose of producing drugs for prophylactic treatment or therapeutic intervention of NLRP3-mediated disorders.

[0188] In another embodiment, the disclosure relates to a method for treating an NLRP3 inflammasome-mediated inflammatory disease or disorder by administering a therapeutically effective amount of a compound of structure (I) to a patient in need of treatment.

[0189] In one embodiment, diseases that can be treated with a compound of structure (I) include type II diabetes, atherosclerosis, Alzheimer's disease, aging, fatty liver, metabolic syndrome, asthma, psoriasis, obesity, acute and chronic tissue injury due to infection, gout, arthritis, enteritis, hepatitis, peritonitis, silicosis, sunburn due to UV radiation, contact hypersensitivity, sepsis, cancer, neurodegenerative diseases, multiple sclerosis, and Mackle-Wells syndrome.

[0190] In one other embodiment, the compound of structure (I) is used in a method for treating a disorder or disease selected from autoimmune disorders, inflammatory disorders, cardiovascular diseases, neurodegenerative disorders, bacterial and viral infections, allergies, asthma, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, transplantation, sperm motility, red blood cell deficiency, graft rejection, lung disorders, respiratory diseases, ischemic conditions, and cancer.

[0191] In some embodiments, the NEK7-related disorders treatable with compounds of structure (I) are selected from rheumatoid arthritis, psoriatic arthritis, osteoarthritis, systemic lupus erythematosus, lupus nephritis, ankylosing spondylitis, osteoporosis, systemic sclerosis, multiple sclerosis, psoriasis, type 1 diabetes, type 2 diabetes, inflammatory bowel disease (Crohn's disease and ulcerative colitis), hyperimmune globulin D emia and periodic fever syndromes, cryopyrin-associated periodic fever syndromes, Schnitzler syndrome, systemic juvenile idiopathic arthritis, adult-onset Still's disease, gout, pseudogout, SAPHO syndrome, Castleman disease, sepsis, stroke, atherosclerosis, celiac disease, DIRA (IL-I receptor antagonist deficiency), Alzheimer's disease, Parkinson's disease, and cancer.

[0192] Therapeutic methods in which a compound of at least one structure (I) is administered in combination with an anti-inflammatory or therapeutic agent are also included herein. Anti-inflammatory agents include, but are not limited to, NSAIDs, nonspecific and COX-2 specific cyclooxygenase enzyme inhibitors, gold compounds, corticosteroids, methotrexate, tumor necrosis factor (TNF) antagonists, immunosuppressants, and methotrexate. Examples of NSAIDs include, but are not limited to, ibuprofen, flurbiprofen, naproxen and naproxen sodium, diclofenac, combinations of diclofenac sodium and misoprostol, sulindac, oxaprozin, diflunisal, piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen, sodium nabumetone, sulfasalazine, tolmetin sodium, and hydroxychloroquine.

[0193] Other examples of NSAIDs include COX-2 specific inhibitors, such as celecoxib, valdecoxib, lumiracoxib, and / or etoricoxib.

[0194] In some embodiments, the anti-inflammatory agent is a salicylate. Examples of salicylates include, but are not limited to, acetylsalicylic acid, i.e., aspirin, sodium salicylate, and choline and magnesium salicylate.

[0195] Anti-inflammatory drugs may also be corticosteroids. For example, corticosteroids may include cortisone, dexamethasone, methylprednisolone, prednisolone, prednisolone sodium phosphate, or prednisone.

[0196] In further embodiments, the anti-inflammatory agent is a gold compound, such as sodium goldthiomalate or auranofin.

[0197] The disclosure also includes embodiments in which the anti-inflammatory agent is a metabolic inhibitor, such as a dihydrofolate reductase inhibitor (e.g., methotrexate) or a dihydroorotate dehydrogenase inhibitor (e.g., leflunomide).

[0198] Therapeutic agents include drugs for pain and inflammation, such as histamine and histamine antagonists, bradykinin and bradykinin antagonists, 5-hydroxytryptamine (serotonin), lipid substances produced by the biotransformation of products of selective hydrolysis of membrane phospholipids, eicosanoids, prostaglandins, thromboxanes, leukotrienes, aspirin, nonsteroidal anti-inflammatory drugs, analgesics and antipyretics, drugs that inhibit the synthesis of prostaglandins and thromboxanes, and selective agents for inducible cyclooxygenase. Inhibitors, selective inhibitors of inducible cyclooxygenase-2, autacoids, paracrine hormones, somatostatins, gastrin, cytokines that mediate interactions involved in humoral and cellular immune responses, lipid-derived autacoids, eicosanoids, β-adrenergic agonists, ipratropium, glucocorticoids, methylxanthines, sodium channel blockers, opioid receptor agonists, calcium channel blockers, membrane stabilizers, and leukotriene inhibitors may also be included.

[0199] Other embodiments of the present disclosure relate to combinations in which at least one anti-inflammatory compound is an anti-monoclonal antibody (e.g., eculizumab or paxerizumab), a TNF antagonist (e.g., etanercept), or infliximab (an anti-TNF alpha monoclonal antibody).

[0200] Therapeutic agents used in combination with the compound of structure (I) may also include small molecule compounds that inhibit the activation of the NLRP3 inflammasome, such as MCC950, sulforaphane, isoliquiritigenin, β-hydroxybutyrate, flufenamic acid, mefenamic acid, 3,4-methylenedioxy-β-nitrostyrene (MNS), and parthenolide.

[0201] Further embodiments of the present disclosure relate to combinations in which at least one active agent is an immunosuppressant compound, such as an immunosuppressant compound selected from methotrexate, leflunomide, cyclosporine, tacrolimus, azathioprine, and mycophenolate mofetil.

[0202] The compound of the disclosed structure (I) may be administered in combination with other known therapeutic agents (such as anticancer drugs). As used herein, the term “anticancer drug” refers to any drug administered to a cancer patient for the purpose of treating cancer.

[0203] In some embodiments, the anticancer drug belongs to the following category. Alkylating agents: e.g., altoretamine, bendamustine, busulfan, carmustine, chlorambucil, chlormethine, cyclophosphamide, dacarbazine, ifosfamide, improsulfan, tosylate, lomustine, melphalan, mitobronitol, mitractol, nimustine, ranimustine, temozolomide, thiotepa, treosulfan, mechloretamine, carbocon; apadicone, fotemustine, glucosphamide, paliphosphamide, pipobromane, trophosphamide, uramustine, TH-3024, VAL-0834, etc. Platinum compounds: for example, carboplatin, cisplatin, eptaplatin, miriplatin hydrate, oxaliplatin, lovaplatin, nedaplatin, picoplatin, satraplatin; lovaplatin, nedaplatin, picoplatin, satraplatin, etc. DNA agonists: for example, amrubicin, bisanthren, decitabine, mitoxantrone, procarbazine, trabectedin, clofarabine; amsacrin, brostaricin, picantrone, laromustine 1,3, etc. Topoisomerase inhibitors: e.g., etoposide, irinotecan, razoxane, sobuzoxane, teniposide, topotecan; amonafide, berotecan, eriptinium acetate, boreroxine, etc. Microtubule polymerization regulators: e.g., cabazitaxel, docetaxel, eribulin, ixabepyrone, paclitaxel, vinblastine, vincristine, vinorelbine, vindesine, vinflunin; phosbletabrin, tesetaxel, etc. Antimetabolites: e.g., asparaginase 3, azacitidine, levofolinate calcium, capecitabine, cladribine, cytarabine, enocitabine, phloxuridine, fludarabine, fluorouracil, gemcitabine, mercaptopurine, methotrexate, nelarabine, pemetrexed, pralatrexate, azathioprine, thioguanine, carmofur; doxyfluridine, elacitarabine, larcitrexed, sapacitabine, tegafur 2,3, trimethrexate, etc. Anticancer antibiotics: for example, bleomycin, dactinomycin, doxorubicin, epirubicin, idarubicin, rebamisol, miltefosine, mitomycin C, romidepsin, streptozocin, barurubicin, dinostatin, zolubicin, daunurobicin, plicamycin; acralubicin, peplomycin, pirarubicin, etc. Hormones / antagonists: e.g., abarelix, abiraterone, bicalutamide, buserelin, carsterone, chlorotrianicene, degarelix, dexamethasone, estradiol, fluocortone, fluoxymesterone, flutamide, fulvestrant, goserelin, histrelin, leuprorelin, megestrol, mitotane, nafarelin, nandrolone, nilutamide, octreotide, prednisolone, raloxifene, tamoxifen, thyrotropin alpha, toremifene, trilostane, triptorelin, diethylstilbestrol; acorbifen, danazol, deslorerin, epithiostanol, orteronel, enzalutamide 1,3, etc. Aromatase inhibitors: e.g., aminoglutethimide, anastrozole, exemestane, fadrozole, letrozole, testolactone; formestane, etc. Small molecule kinase inhibitors: e.g., crizotinib, dasatinib, erlotinib, imatinib, lapatinib, nilotinib, pazopanib, regorafenib, ruxolitinib, sorafenib, sunitinib, vandetanib, vemurafenib, bosutinib, gefitinib, axitinib; afatinib, arisertib, dabrafenib, dacomitinib, dinaciclib, dovicinib B, Enzastaurin, Nintedanib, Lenvatinib, Linifanib, Lincitinib, Masitinib, Midostaurin, Motesanib, Neratinib, Orantinib, Perifosin, Ponatinib, Radotinib, Rigosatib, Tipifanib, Tivantinib, Tivozanib, Trametinib, Pimasertib, Brivanib Alanine Ester, Cejilanib, etc.

[0204] In some embodiments, the drugs administered in combination with the compounds described herein include any suitable drugs that are usefully delivered by inhalation, such as analgesics (e.g., codeine, dihydromorphine, ergotamine, fentanyl, or morphine), angina preparations (e.g., diltiazem), antiallergic drugs (e.g., cromoglycate, ketotifen, or nedocromil), antiinfective drugs (e.g., cephalosporins, penicillin, streptomycin, sulfonamides, tetracycline, or pentamidine), antihistamines (e.g., metapirylene), anti-inflammatory drugs (e.g., beclomethasone, flunisolide, budesonide, tipredane, triamcinolone acetonide, or fluticasone), antitussives (e.g., noscapine), and bronchodilators (e.g., ephedra). Drine, adrenaline, fenoterol, formoterol, isoprenaline, metaproterenol, phenylephrine, phenylpropanolamine, pyrbuterol, reproterol, limiterol, salbutamol, salmeterol, terbutaline, isoethaline, tulobuterol, orciprenaline or (-)-4-amino-3,5-dichloro-α-[[[6-[2-(2-pyridinyl)ethoxy]hexyl]-amino]methyl]benzenemethanol), diuretics (e.g., amiloride), anticholinergics (e.g., ipratropium, atropine or oxytropium), hormones (e.g., cortisone, hydrocortisone or prednisolone), xanthines (e.g., aminophylline, choline theophylline, lysine theophylline) These include theophyllinates (or theophylline), as well as therapeutic proteins and peptides (e.g., insulin or glucagon). It will be apparent to those skilled in the art that, where appropriate, the drugs are used in the form of salts (e.g., as alkali metal or amine salts, or as acid addition salts), as esters (e.g., lower alkyl esters), or as solvates (e.g., hydrates) to optimize the activity and / or stability of the drugs.

[0205] The agents disclosed herein or other suitable agents are administered according to the disease being treated. Therefore, in some embodiments, one or more of the compounds of this disclosure will be co-administered with other agents such as those described above. When used in combination therapy, the compounds of this disclosure are administered simultaneously with or separately from a second agent. This combination administration may include administering the two agents simultaneously in the same dosage form, simultaneously in separate dosage forms, and separately. That is, either the compounds of this disclosure or any of the agents described above may be formulated together in the same dosage form and administered simultaneously. Alternatively, either the compounds of this disclosure or any of the agents described above may be present in separate formulations and administered simultaneously. In another alternative, either the compound of this disclosure or any of the agents described above may be administered immediately afterward, and vice versa. In some embodiments of the separate administration protocol, either the compound of this disclosure or any of the agents described above may be administered with a delay of several minutes, several hours, or several days.

[0206] In some embodiments, the compound of structure (I) is administered as monotherapy.

[0207] For the identification of signal transduction or mechanistic pathways, and for the detection of interactions between various signal transduction pathways, scientists have developed suitable models or model systems, such as cell culture models and transgenic animal models. To quantify specific stages of a signal transduction cascade, interacting compounds can be used to modulate the signal. The compounds of the embodiments of the present invention can also be used as reagents for examining NEK7-dependent signal transduction pathways in animal and / or cell culture models, or in clinical diseases as referred to in this application.

[0208] The methods of the embodiments of the present invention can be carried out either in vitro or in vivo. The sensitivity of specific cells to treatment with the compound of structure (I) can be determined, in particular, by in vitro testing, whether in the course of research or clinical application. Generally, cell cultures are combined with various concentrations of the compound for a period of time sufficient for the active agent to inhibit NEK7 activity, usually from about 1 hour to 1 week. In vitro treatment can be performed using biopsy samples or cultured cells derived from cell lines.

[0209] In some embodiments, IC of a compound with structure (I) that inhibits NEK7 50 This was determined by the concentration of the compound required to inhibit 50% of the activity of NEK kinase. The compound of structure (I) was used in concentrations of less than about 5 mM, preferably less than about 1 mM, and more preferably less than about 0.100 mM, as described in more detail in the examples. 50 The efficacy value was shown.

[0210] The examples and preparations described below further illustrate and demonstrate the compounds of this disclosure and the methods for preparing and testing such compounds. It should be understood that the scope of this disclosure is not limited in any way by the scope of the following examples and preparations. In the following examples, and throughout the specification and claims, molecules having only one stereocenter exist as racemic mixtures unless otherwise specified. Molecules having two or more stereocenters exist as racemic mixtures of diastereomers unless otherwise specified. Single enantiomers / diastereomers may be obtained by methods known to those skilled in the art. [Examples]

[0211] The following examples are provided for illustrative purposes only.

[0212] General procedure All proton NMR experiments were recorded at 400 MHz using a Bruker NEO Spectrometer equipped with a BBFO probe. The deuterated solvent contained less than 0.05% v / v of tetramethylsilane (set at 0.00 ppm) which was used as a reference signal. If the deuterated solvent did not contain tetramethylsilane, the peak of the remaining non-deuterated solvent was used as the reference signal according to the published guidelines (J. Org. Chem. 1997, 62(21), 7512-7515). Chemical shifts are expressed in parts per million (ppm, in δ units). Coupling constants are in Hertz (Hz). Splitting patterns describe the apparent multiplicity and are expressed as s (singular), d (double), t (tripular), q (quadular), m (multiple), qt (quintular), or brs (broad single). LC / MS analysis was performed using an Agilent Technologies UHPLC 1290 Infinity II with a G6125 MS detector. The microwave reaction was performed using a standard protocol on an Anton Paar GmbH Monowave 300.

[0213] NEK7 Enzyme Assay Casein substrate (milk-derived, a mixture of hydrolyzed and partially dephosphorylated α, β, and κ caseins, obtained from Sigma Aldrich, catalog #C4765, diluted to a final concentration of 1 mg / mL with distilled water) and fully recombinant human NEK7 (expressed by baculovirus in Sf9 insect cells with an N-terminal GST tag, obtained from SignalChem, catalog #N09-10G, 0.1 μg / μL) were mixed in assay buffer (20 mM Hepes pH 7.5, 10 mM MgCl2, 1 mM EGTA, 0.02% Brij35, 0.02 mg / mL BSA, 0.1 mM Na3VO4, 2 mM DTT, 1% DMSO). The compound of interest (stepwise 3-fold dilution with DMSO from 10 μM to 0.5 nM) or Vehicle (1% DMSO) was partitioned into the kinase reaction mixture using acoustic technology (Echo550; nanoliter range). After incubating at room temperature for 20 minutes, [33 The kinase reaction was initiated by adding [P]-ATP (specific activity 10 μCi / μl), and the mixture was incubated at room temperature for 2 hours. Next, the reaction was stopped by spotting the reaction mixture onto phosphocellulose P81 paper. After washing, the radioactivity of the P81 paper was measured, and the kinase activity data was expressed as a percentage of the residual kinase activity in the test sample compared to the vehicle reaction product. IC was calculated using Prism (GraphPad Software). 50 Values ​​and curve fit were obtained. IL-1β release assay Approximately 1.5 million THP-1 cells were seeded in each well of a 6-well TC plate and incubated with 40 nM PMA in RPMI (10% FBS, 1% Penstrep) for 24 hours. Next, the culture medium was removed and the cells rested in RPMI (10% FBS, 1% Penstrep) for 24 hours. After removing the culture medium, the cells were pretreated with various concentrations of the compound of interest in RPMI (5% FBS) for 2 hours (typically serially diluted 3-fold in RPMI + 5% FBS, from 1 μM to 0.5 nM). The culture medium was removed again and the cells were incubated with 250 ng / mL LPS and the compound of interest (concentrations as described above) in RPMI (5% FBS) for 2 hours. Finally, the culture medium was removed and the cells were incubated with 20 μM nigericin and the compound of interest (concentrations as described above) in Opti-MEM for 30 minutes. Next, the cell culture medium was collected, and the amount of cleaved IL-1β was determined using a JESS instrument (Protein Simple) and standard protocol. Cleaved IL-1β antibody was obtained from Cell Signaling (catalog #83186S) and used at a 1:20 dilution in antibody diluent 2. For chemiluminescent detection, Protein Simple 1x anti-Rabbit HRP secondary antibody was used along with Protein Simple luminol and peroxide. The primary antibody incubation time was increased from 30 minutes to 60 minutes.

[0214] Abbreviation °C (degrees Celsius); 1¹H NMR (Proton Nuclear Magnetic Resonance); ACN (Acetonitrile); Boc (Tert-Butyloxycarbonyl); DCM (Dichloromethane); DIPEA (N,N-Diisopropylethylamine); DMAP (4-Dimethylaminopyridine); DMF (N,N-Dimethylformamide); DMSO-d6 (Deuterated Dimethyl Sulfoxide); eq (Equivalent); Â (Ethyl Acetate); g (Gram); (g) Gas; h (Hour); HPLC (High-Performance Liquid Chromatography); LCMS (Liquid Chromatography Mass Spectrometry); MeOH (Methanol); mg (Milligram); min (Minute); mL (Milliliter) );mmol (millimole);N (n);n-BuOH (1-butanol);Pd(PPh3)4 (palladium-tetrakis(triphenylphosphine));PdCl2(dppf)([1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride);sec- (secondary);TBAF (tetra-n-butylammonium fluoride);TBDMS (tert-butyldimethylsilyl);tert- (tertiary);TFA (trifluoroacetic acid);THF (tetrahydrofuran);TLC (thin-layer chromatography);UPLC (ultrafast performance liquid chromatography)

[0215] Preparation of synthetic intermediates [ka] Copper(II) acetate (0.348 g, 1.916 mmol), 2,2'-bipyridine (0.299 g, 1.916 mmol), and sodium bicarbonate (0.322 g, 3.830 mmol) were added to a stirred solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.500 g, 1.916 mmol) and cyclopropylboronic acid (0.329 g, 3.830 mmol) in dichloroethane (10 mL). The resulting mixture was stirred at 70°C under an oxygen atmosphere for 12 hours. After the reaction was complete (shown by TLC), the reaction mixture was filtered through a diatomaceous earth pad and then rinsed with DCM (20 mL × 2). The combined filtrate was washed with water (20 mL) and brine (25 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. This was purified by flash chromatography (silica gel, 230-400 mesh, in petroleum ether, eluted with 20% siRNA) to obtain the title compound as an off-white solid (0.24 g, yield 36%). 1 H NMR (400 MHz, DMSO-d6) δ = 8.21 (s, 1H), 3.74-3.79 (m, 1H), 1.11-1.15 (m, 2H), 1.04-1.09 (m, 2H).LCMS: 301.8 [M+H]

[0216] [ka] Cs2CO3 (12.38 g, 38.31 mmol) and 2-iodopropane (3.60 g, 21.16 mmol) were added to a stirred solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (5.00 g, 19.15 mmol) in DMF (25 mL) in a sealed 25 mL tube. The reaction mixture was stirred at 90°C for 16 hours. After the reaction was complete (shown by TLC), the mixture was poured over crushed ice (50 g) and stirred for 15 minutes. The resulting solid was filtered, washed with water (2 × 5 mL), and dried to obtain the title compound as an off-white solid (3.25 g, 56% yield). 1H NMR (400 MHz, DMSO-d6) δ = 8.18 (s, 1H), 4.93-4.99 (m, 1H), 1.42 (d, J = 6.8 Hz, 6H). LCMS: 303.8 [M+H]

[0217] Intermediate A3 to A12 The following intermediates were prepared using the same procedure as described for intermediate A2 (replacing 2-iodopropane with the appropriate reagent shown below (alkyl halide or tosylate)). [Table 24] [Table 25]

[0218] [ka] NaH2PO4 (0.044 g, 0.372 mmol) was added to a mixture of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.100 g, 0.380 mmol), 2,2-dimethyloxirane (0.055 g, 0.760 mmol), and K2CO3 (0.050 g, 0.372 mmol) in acetonitrile (3 mL) and water (1 mL). The resulting solution was subjected to microwave irradiation at 150°C for 1 hour. After the reaction was complete (shown by TLC), the solvent was removed under reduced pressure to obtain the crude product, which was purified by flash chromatography (silica gel, 230-400 mesh, in petroleum ether, eluted with 25% siRNA) to obtain the title compound as a light brown solid (0.064 g, yield 51%). 1 H NMR (400 MHz, DMSO-d6) δ = 8.20 (s, 1H), 4.19 (s, 2H), 1.09 (s, 6H).LCMS: 334.0 [M+H]

[0219] [ka] Starting with 3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (1.00 g, 3.83 mmol) and pyridine-4-ylboronic acid (0.94 g, 7.66 mmol), the title compound was prepared following the same procedure as described for intermediate A1, and obtained as a light brown solid (0.27 g, 21% yield). LCMS: 338.8 [M+H]

[0220] [ka] Cs2CO3 (0.780 g, 2.394 mmol) and methyl iodide (0.138 mL, 2.203 mmol) were added to a solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.500 g, 1.916 mmol) in DMF (3 mL) (at 0°C, in a sealed 25 mL tube). The reaction mixture was stirred at 25°C for 1 hour. After the reaction was complete (shown by TLC), the mixture was poured over crushed ice (50 g) and stirred for 30 minutes. The resulting solid was filtered, washed with water (2 × 5 mL), and dried to obtain the title compound as a yellow solid (0.380 g, 67% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.22 (s, 1H), 6.79 (s, 2H), 3.89 (s, 3H).LCMS: 276.0 [M+H]

[0221] [ka] KOH (1.320 g, 23.0 mmol) and iodine (1.620 g, 12.8 mmol) were added to a solution of 4-chloro-1H-pyrazolo[4,3-c]pyridine (1.000 g, 6.4 mmol) in dioxane (10 mL), and the resulting mixture was stirred at 75°C for 4 hours. After the reaction was complete (shown by TLC), the reaction mixture was filtered through a diatomaceous earth pad, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by reverse-phase column chromatography to obtain the title compound as a white solid (0.633 g, yield 63%). 1H NMR (400 MHz, DMSO-d6) δ = 14.12 (bs, 1H), 8.14 (d, J = 6.0 Hz, 1H), 7.66 (d, J = 5.6 Hz, 1H).LCMS: 279.9 [M+H]

[0222] [ka] Starting with 4-chloro-3-iodo-1H-pyrazolo[4,3-c]pyridine (0.630 g, 2.20 mmol) and cyclopropylboronic acid (0.329 g, 3.83 mmol), the title compound was prepared according to the same procedure as described for intermediate A1 and obtained as a white solid (0.430 g, 60% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.21 (d, J = 6.0 Hz, 1H), 7.81 (d, J = 5.6 Hz, 1H), 3.84-3.89 (m, 1H), 1.14-1.17 (m, 4H).LCMS: 319.7 [M+H]

[0223] [ka] A mixture of 4-chloro-1-cyclopropyl-3-iodo-1H-pyrazolo[4,3-c]pyridine (0.20 g) and an aqueous solution of ammonium hydroxide (25% aqueous solution, 8 mL) was subjected to microwave irradiation at 150°C for 2 hours. After the reaction was complete (shown by TLC), the reaction mixture was concentrated under reduced pressure to obtain the title compound as an off-white solid (0.19 g, quantitative yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.11 (s, 1H), 7.38 (s, 1H), 6.68 (bs, 2H), 3.48-3.54 (m, 1H), 0.97-0.99 (m, 4H).LCMS: 301.0 [M+H]

[0224] [ka] 3-(benzyloxy)cyclobutyl methanesulfonate (prepared as described in PCT publication number WO2019 / 092170, 0.491 g, 1.916 mmol) and Cs2CO3 (0.624 g, 1.916 mmol) were added to a solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.250 g, 0.958 mmol) in DMF (5 mL), and the resulting mixture was stirred at 90°C for 12 hours. After the reaction was complete (shown by TLC), the reaction mixture was poured into ice water (50 mL) and extracted with ethyl acetate (2 × 30 mL). The combined organic layer was dried over Na2SO4, filtered, and evaporated under reduced pressure to obtain the title compound as a light brown gum (0.400 g, 75% purity LCMS), which was used without further purification. LCMS: 422.0 [M+H]

[0225] [ka] The title compound was prepared as described in PCT publication number WO2002 / 076986.

[0226] [ka] A mixture of 4-chloro-1-cyclopropyl-3-iodo-1H-pyrazolo[4,3-c]pyridine (A16, 0.880 g, 2.75 mmol) and (2,5-dimethoxyphenyl)methaneamine (1.245 mL, 8.26 mmol) in n-BuOH (10 mL) was stirred at 110°C for 12 hours. After the reaction was complete (shown by LC-MS), the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified using Isolera (silica gel, 230-400 mesh, in petroleum ether, eluted with 40% ethyl acetate) to obtain the title product as a yellow gum (1.0 g, yield 80%). 1H NMR (400 MHz, DMSO-d6) δ = 7.81 (d, J = 6.4 Hz, 1H), 7.19 (d, J = 8.4 Hz, 1H), 6.87 (d, J = 6.0 Hz, 1H), 6.60 (d, J = 2.4 Hz, 1H), 6.53-6.56 (m, 1H), 6.45-6.47 (m, 1H), 4.62 (d, J = 5.6 Hz, 2H), 3.88 (s, 3H), 3.73 (s, 3H), 3.66-3.70 (m, 1H), 1.01-1.10 (m, 4H).LCMS: 451.0 [M+H]

[0227] [ka] The title compound was prepared as described in PCT publication number WO2011 / 119663.

[0228] [ka] K2CO3 (0.397 g, 2.870 mmol) and 1-bromo-2-methoxyethane (0.319 g, 2.299 mmol) were added to a solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.500 g, 1.916 mmol) in DMF (6 mL), and the resulting mixture was stirred in a sealed tube at 80°C for 12 hours. After the reaction was complete (shown by TLC), the reaction mixture was poured over crushed ice (25 g) and extracted with SiO2CO3 (2 × 50 mL). The combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the title product (0.500 g, unpurified), which was used without further purification. 1 H NMR (400 MHz, DMSO-d6) δ = 8.20 (s, 1H), 4.43 (t, J = 5.6 Hz, 2H), 3.75 (t, J = 5.6 Hz, 2H), 3.20 (s, 3H).LCMS: 319.8 [M+H]

[0229] [ka] The title compound was prepared as described in USPTO publication number WO2015 / 165279.

[0230] [ka] The title compound was prepared as described in USPTO publication number WO2018 / 121228.

[0231] [ka] A mixture of 1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A1, 0.500 g, 1.66 mmol), 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.433 g, 1.82 mmol), and K2CO3 (0.688 g, 4.98 mmol) in 1,4-dioxane (25 mL) and water (2.5 mL) was purged with N2 for 10 minutes. Then Pd(PPh3)4 (0.092 g, 0.08 mmol) was added, and the reaction mixture was stirred at 100°C for 16 hours. After the reaction was complete (shown by TLC), the mixture was filtered through a diatomaceous earth pad and then rinsed with ELISA (2 × 10 mL). The combined filtrate was concentrated under reduced pressure to obtain a crude substance, which was purified by flash chromatography (silica gel, 230-400 mesh, in DCM, eluted with 2% MeOH) to obtain the title compound as a yellow solid (0.46 g, yield 98%). 1 H NMR (400 MHz, DMSO-d6) δ = 8.23 ​​(s, 1H), 7.15-7.24 (m, 2H), 6.87-6.91 (m, 1H), 5.47 (bs, 2H), 3.80-3.84 (m, 1H), 1.18-1.19 (m, 2H), 1.05-1.08 (m, 2H).LCMS: 285.0 [M+H]

[0232] [ka] Starting with 3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A2, 1.887 g, 6.23 mmol) and (4-nitrophenyl)boronic acid (1.56 g, 9.34 mmol), the title compound was prepared according to the same procedure as described for intermediate B1, and obtained as a yellow solid (1.242 g, 67% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.38-8.40 (m, 2H), 8.28 (s, 1H), 7.92-7.95 (m, 2H), 5.07-5.14 (m, 1H), 1.51 (d, J = 6.8 Hz, 6H).LCMS: 299.1 [M+H]

[0233] [ka] Iron powder (2.320 g, 41.60 mmol) and ammonium chloride (2.220 g, 41.60 mmol) were added to a stirred solution of 1-isopropyl-3-(4-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (1.242 g, 4.16 mmol) in ethanol (50 mL) and water (20 mL). The resulting mixture was heated at 80°C for 3 hours. After the reaction was complete (shown by TLC), the reaction mixture was filtered through a diatomaceous earth pad and then rinsed with SiO2 (2 × 25 mL). The combined filtrate was concentrated under reduced pressure, the residue was dissolved in SiO2 (100 mL), washed with brine (25 mL), dried over Na2SO4, filtered, and evaporated under reduced pressure to obtain the title compound as a pale yellow solid (1.042 g, quantitative yield), which was left unpurified.

[0234] [ka] Starting with 3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A2, 0.10 g, 0.32 mmol) and (3-fluoro-4-nitrophenyl)boronic acid (0.71 g, 0.39 mmol), the title compound was prepared following the same procedure as described for intermediate B1, and obtained as a yellow solid (0.07 g, 67% yield). LCMS: 315.1 [MH]

[0235] [ka] Starting with 3-(3-fluoro-4-nitrophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.07 g, 0.22 mmol) and Fe / NH4Cl, the title compound was prepared following a procedure similar to that described for step 2 of intermediate B2, and obtained as a pale yellow solid (0.09 g, quantitative yield), which was not further purified. LCMS: 287.1 [M+H]

[0236] [ka] Starting with 3-iodo-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A3, 0.800 g, 2.522 mmol) and (4-nitrophenyl)boronic acid (0.632 g, 3.78 mmol), the title compound was prepared according to the same procedure as described for intermediate B1, and obtained as a yellow solid (0.596 g, 76% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.41-8.43 (m, 2H), 8.30 (s, 1H), 7.99-8.01 (m, 2H), 6.05-6.08 (m, 1H), 4.97-5.12 (m, 4H).LCMS: 311.0 [MH]

[0237] [ka] Starting with 3-(4-nitrophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.596 g, 1.91 mmol) and Fe / NH4Cl, the title compound was prepared following a procedure similar to that described for step 2 of intermediate B2, and obtained as a pale yellow solid (0.42 g, quantitative yield), which was not further purified. LCMS: 283.0 [M+H]

[0238] [ka] Starting with 3-iodo-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A3, 0.110 g, 0.346 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.099 g, 0.420 mmol), the title compound was prepared following a procedure similar to that described for intermediate B1, and obtained as a pale yellow gum (0.077 g, 74% yield). LCMS: 317.1 [M+H]

[0239] [ka] Starting with 1-allyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A4, 0.15 g, 0.49 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.13 g, 0.54 mmol), the title compound was prepared following a procedure similar to that described for intermediate B1, and obtained as a yellow gum (0.13 g, 92% yield). LCMS: 285.0 [M+H]

[0240] [ka] Starting with 1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A1, 0.110 g, 0.36 mmol) and (4-nitrophenyl)boronic acid (0.067 g, 0.40 mmol), the title compound was prepared according to the same procedure as described for intermediate B1, and obtained as a pale yellow solid (0.060 g, 56% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.37 (d, J = 8.8 Hz, 2H), 8.30 (s, 1H), 7.91 (d, J = 8.8 Hz, 2H), 3.91-3.94 (m, 1H), 1.23-1.24 (m, 2H), 1.11-1.14 (m, 2H).LCMS: 297.0 [M+H]

[0241] [ka] Starting with 1-cyclopropyl-3-(4-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.060 g, 0.2 mmol) and Fe / NH4Cl, the title compound was prepared following a procedure similar to that described for step 2 of intermediate B2, and obtained as a pale yellow solid (0.047 g, 89% yield), which was not further purified. LCMS: 266.9 [M+H]

[0242] [ka] Starting with 1-cyclobutyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A5, 0.150 g, 0.47 mmol) and (4-nitrophenyl)boronic acid (0.087 g, 0.52 mmol), the title compound was prepared according to the same procedure as described for intermediate B1, and obtained as a pale yellow solid (0.160 g, quantitative yield). 1H NMR (400 MHz, DMSO-d6) δ = 8.34-8.41 (m, 2H), 8.28 (s, 1H), 7.94-7.97 (m, 2H), 5.32-5.43 (m, 1H), 2.65-2.75 (m, 2H), 1.86-1.94 (m, 4H).LCMS: 311.2 [M+H]

[0243] [ka] Starting with 1-cyclobutyl-3-(4-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.16 g, 0.51 mmol) and Fe / NH4Cl, the title compound was prepared following a procedure similar to that described for step 2 of intermediate B2, obtained as a pale yellow solid (0.140 g, quantitative yield), and the process proceeded without further purification. LCMS: 281.0 [M+H]

[0244] [ka] Starting with 1-cyclobutyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A5, 0.250 g, 0.793 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.225 g, 0.952 mmol), the title compound was prepared following the same procedure as described for intermediate B1, and obtained as a pale yellow solid (0.100 g, 32% yield). LCMS: 299.1 [M+H]

[0245] [ka] Starting with 3-iodo-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A6, 0.165 g, 0.49 mmol) and (4-nitrophenyl)boronic acid (0.091 g, 0.54 mmol), the title compound was prepared according to the same procedure as described for intermediate B1, and obtained as a pale yellow solid (0.086 g, 53% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.38-8.40 (m, 2H), 8.30 (s, 1H), 7.93-7.95 (m, 2H), 5.52-5.58 (m, 1H), 4.06-4.15 (m, 2H), 3.88-3.99 (m, 2H), 2.50-2.51 (m, 2H).LCMS: 327.2 [M+H]

[0246] [ka] Starting with 3-(4-nitrophenyl)-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.115 g, 0.35 mmol) and Fe / NH4Cl, the title compound was prepared following a procedure similar to that described for step 2 of intermediate B2, and obtained as a pale yellow solid (0.083 g, 80% yield), which was then removed without further purification. 1 H NMR (400 MHz, DMSO-d6) δ = 8.22 (s, 1H), 7.33 (dd, J = 2.0, 6.6 Hz, 2H), 6.71 (dd, J = 2.0, 6.4 Hz, 2H), 5.48 (bs, 2H), 5.44-5.47 (m, 1H), 4.04-4.12 (m, 2H), 3.86-3.94 (m, 2H), 2.34-2.41 (m, 2H).LCMS: 296.9 [M+H]

[0247] [ka] Starting with 3-iodo-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A6, 0.351 g, 1.00 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.301 g, 1.27 mmol), the title compound was prepared according to the same procedure as described for intermediate B1, and obtained as a pale yellow solid (0.200 g, 60% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.23 ​​(s, 1H), 7.24-7.28 (m, 1H), 7.18-7.21 (m, 1H), 6.88-6.92 (m, 1H), 5.46-5.50 (m, 3H), 4.04-4.12 (m, 2H), 3.87-3.94 (m, 2H), 2.33-2.41 (m, 2H).LCMS: 315.1 [M+H]

[0248] [ka] Starting with 3-iodo-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A7, 0.094 g, 0.273 mmol) and (4-nitrophenyl)boronic acid (0.055 g, 0.328 mmol), the title compound was prepared according to the same procedure as described for intermediate B1, and obtained as a pale yellow solid (0.078 g, 84% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.37-8.40 (m, 2H), 8.29 (s, 1H), 7.93-7.96 (m, 2H), 4.96-5.02 (m, 1H), 4.00-4.04 (m, 2H), 3.54-3.59 (m, 2H), 2.16-2.24 (m, 2H), 1.89-1.94 (m, 2H).LCMS: 340.9 [M+H]

[0249] [ka] Starting with 3-(4-nitrophenyl)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.078 g, 0.23 mmol) and Fe / NH4Cl, the title compound was prepared following a procedure similar to that described for step 2 of intermediate B2, and obtained as a pale yellow solid (0.052 g, 73% yield), which was not further purified. LCMS: 311.1 [M+H]

[0250] [ka] Starting with 3-iodo-1-(tetrahydro-2H-pyran-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A8, 0.050 g, 0.14 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.040 g, 0.17 mmol), the title compound was prepared following a procedure similar to that described for intermediate B1, and obtained as a yellow gum (0.015 g, 32% yield). LCMS: 329.2 [M+H]

[0251] [ka] Starting with 1-(3,3-difluorocyclobutyl)-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A9, 0.100 g, 0.285 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.810 g, 0.341 mmol), the title compound was prepared following the same procedure as described for intermediate B1, and obtained as a pale yellow solid (0.088 g, 93% yield). LCMS: 334.9 [M+H]

[0252] [ka] An aqueous solution of HCl (1.5 N, 5 mL) was added to a solution of 3-iodo-1-(1,4-dioxaspiro[4.5]decane-8-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A10, 0.313 g, 0.780 mmol), and the resulting mixture was stirred at room temperature for 12 hours. After the reaction was complete (shown in UPLC), the solvent was removed under reduced pressure, and the residue (0.232 g, LCMS: 358.0 [M+H]) was obtained and dissolved in THF (5 mL). The resulting solution was cooled to 0°C, NaBH4 (0.050 g, 1.322 mmol) was added, and the mixture was stirred at room temperature for 1 hour. After the reaction was complete (shown in UPLC), an aqueous solution of HCl (1.5 N, 5 mL) was added, and the mixture was extracted with SiO2 (2 × 10 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the title product as a light brown gum (0.196 g). Further purification was not performed. LCMS: 359.8 [M+H]

[0253] [ka] Starting with 4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-1-yl)cyclohexane-1-ol (0.196 g, 0.550 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.160 g, 0.655 mmol), the title compound was prepared following a procedure similar to that described for intermediate B1, and obtained as a yellow gum (0.120 g, 64% yield). LCMS: 343.0 [M+H]

[0254] [ka] Starting with tert-butyl 4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-1-yl)piperidine-1-carboxylate (A11, 0.10 g, 0.23 mmol) and (3-fluoro-4-nitrophenyl)boronic acid (0.05 g, 0.27 mmol), the title compound was prepared following the same procedure as described for intermediate B1, and obtained as a yellow solid (0.04 g, 40% yield). LCMS: 458.1 [MH]

[0255] [ka] TFA (0.5 mL) was added to a solution of tert-butyl 4-(4-amino-3-(3-fluoro-4-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidine-1-yl)piperidine-1-carboxylate (100 mg, 0.218 mmol) in dry DCM (5 mL), and the resulting mixture was stirred at room temperature for 2 hours. After the reaction was complete (shown by ULC), the solvent was evaporated under reduced pressure, and the title product was obtained as a light brown gum (110 mg), which was not further purified. LCMS: 358.1 [MH]

[0256] [ka] Oxetane-3-one (0.020 g, 0.277 mmol) and glacial acetic acid (catalytic amount) were added to a solution of 3-(3-fluoro-4-nitrophenyl)-1-(piperidine-4-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.100 g, 0.280 mmol) in DCM (5 mL), and the resulting mixture was stirred at room temperature for 4 hours. Next, sodium triacetoxyborohydride (0.178 g, 0.840 mmol) was added, and the resulting mixture was stirred at room temperature for 12 hours. After the reaction was complete (shown in ULC), the solution was diluted with DCM (5 mL) and washed with 10% NaHCO3 aqueous solution (5 mL) and brine (5 mL). The organic layer was dried over Na2SO4, filtered, and the solvent was evaporated under reduced pressure to obtain the title product (110 mg), which was then removed without further purification. LCMS: 414.2 [MH]

[0257] [ka] Iron powder (0.135 g, 2.417 mmol) and ammonium chloride (0.142 g, 2.655 mmol) were added to a solution of 3-(3-fluoro-4-nitrophenyl)-1-(1-(oxetan-3-yl)piperidine-4-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.110 g, 0.266 mmol) in ethanol (5 mL) and water (2 mL), and the resulting mixture was stirred at 80°C for 2 hours. After the reaction was complete (shown by TLC), the reaction mixture was filtered through a diatomaceous earth pad and then rinsed with ELISA (5 mL × 2). The combined filtrate was concentrated under reduced pressure to obtain a residue, which was dissolved in EtOAC (10 mL) and then washed with brine (5 mL). The organic layer was separated, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the title compound as an off-white solid (0.05 g, 50% yield). Further purification was not performed. LCMS: 383.9 [M+H]

[0258] [ka] Starting with 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-1-yl)tetrahydrothiophene 1,1-dioxide (A12, 0.08 g, 0.21 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.06 g, 0.25 mmol), the title compound was prepared following a procedure similar to that described for intermediate B1, and obtained as a yellow gum (0.03 g, 40% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.26 (s, 1H), 7.27-7.31 (m, 1H), 7.20-7.23 (m, 1H), 6.88-6.93 (m, 1H), 5.69 (bs, 2H), 5.53 (bs, 2H), 3.72-3.78 (m, 2H), 3.51-3.56 (m, 2H), 2.61-2.70 (m, 2H).LCMS: 362.8 [M+H]

[0259] [ka] Starting with 1-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-1-yl)-2-methylpropan-2-ol (A13, 0.067 g, 0.201 mmol) and (4-nitrophenyl)boronic acid (0.054 g, 0.302 mmol), the title compound was prepared according to the same procedure as described for intermediate B1, and obtained as a yellow solid (0.059 g, 90% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.40 (d, J = 7.6 Hz, 2H), 8.30 (s, 1H), 7.94 (d, J = 6.8 Hz, 2H), 4.32 (bs, 2H), 1.16 (bs, 6H).LCMS: 328.9 [MH]

[0260] [ka] Starting with 1-(4-amino-3-(4-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidine-1-yl)-2-methylpropan-2-ol (0.072 g, 0.219 mmol) and Fe / NH4Cl, the title compound was prepared following a procedure similar to that described for step 2 of intermediate B2, and obtained as a pale yellow solid (0.07 g, quantitative yield), which was not further purified. LCMS: 299.0 [M+H]

[0261] [ka] Starting with 1-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-1-yl)-2-methylpropan-2-ol (A13, 0.110 g, 0.330 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.094 g, 0.396 mmol), the title compound was prepared following the same procedure as described for intermediate B1, and obtained as a pale yellow solid (0.077 g, 66% yield). LCMS: 317.1 [M+H]

[0262] [ka] Starting with 3-iodo-1-(pyridine-4-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A14, 0.250 g, 0.73 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.193 g, 0.81 mmol), the title compound was prepared following a procedure similar to that described for intermediate B1, and obtained as a yellow gum (0.140 g, 59% yield). LCMS: 321.9 [M+H]

[0263] [ka] Starting with 3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A2, 0.115 g, 0.379 mmol) and 2-(3-chloro-4-nitrophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.129 g, 0.455 mmol), the title compound was prepared following a procedure similar to that described for intermediate B1, and obtained as an off-white solid (0.083 g, 66% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.28 (s, 1H), 8.21-8.23 (m, 1H), 7.95-7.95 (m, 1H), 7.82-7.85 (m, 1H), 5.07-5.13 (m, 1H), 1.51 (d, J = 6.4Hz, 6H).LCMS: 332.9 [M+H]

[0264] [ka] Starting with 3-(3-chloro-4-nitrophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (0.083 g, 0.25 mmol) and Fe / NH4Cl, the title compound was prepared following a procedure similar to that described for step 2 of intermediate B2, and obtained as a pale yellow solid (0.900 g, quantitative yield), which was not further purified. LCMS: 302.9 [M+H]

[0265] [ka] Starting with 3-iodo-1-methyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A15, 0.100 g, 0.364 mmol) and 2-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.092 g, 0.364 mmol), the title compound was prepared according to the same procedure as described for intermediate B1 and obtained as a yellow solid (0.094 g, 94% yield). 1H NMR (400 MHz, DMSO-d6) δ =8.23 (s, 1H), 7.45-7.46 (m, 1H), 7.31-7.33 (m, 1H), 6.91-6.93 (m, 1H), 5.68 (bs, 2H), 3.91 (s, 3H).LCMS: 275.0[M+H]

[0266] [ka] Starting with 3-iodo-1-methyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B15, 0.110 g, 0.400 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.104 g, 0.440 mmol), the title compound was prepared following a procedure similar to that described for intermediate B1, and obtained as a brown gum (0.130 g, 97% yield). LCMS: 259.1[M+H]

[0267] [ka] Starting with 1-cyclopropyl-3-iodo-1H-pyrazolo[4,3-c]pyridine-4-amine (A16, 0.190 g, 0.63 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.180 g, 0.75 mmol), the title compound was prepared according to the same procedure as described for intermediate B1 and obtained as a yellow gum (0.070 g, 39% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 7.75-7.76 (m, 1H), 7.21-7.24 (m, 1H), 7.14-7.16 (m, 2H), 6.85-6.87 (m, 1H), 5.78 (bs, 2H), 5.46 (bs, 2H), 3.67-3.69 (m, 1H), 1.09-1.10 (m, 4H).LCMS: 317.1 [M+H]

[0268] [ka] Starting with 1-cyclopropyl-3-iodo-1H-pyrazolo[4,3-c]pyridine-4-amine (A16, 0.190 g, 0.63 mmol) and (4-nitrophenyl)boronic acid (0.126 g, 0.75 mmol), the title compound was prepared according to the same procedure as described for intermediate B1, and obtained as a pale yellow solid (0.090 g, 50% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.36-8.39 (m, 2H), 7.93-7.96 (m, 2H), 7.84 (d, J = 6.0 Hz, 1H), 6.95 (d, J = 6.0 Hz, 1H), 5.98 (bs, 2H), 3.78-3.81 (m, 1H), 1.13-1.16 (m, 4H).LCMS: 296.1 [M+H]

[0269] [ka] Starting with 1-cyclopropyl-3-(4-nitrophenyl)-1H-pyrazolo[4,3-c]pyridine-4-amine (0.093 g, 0.31 mmol) and Fe / NH4Cl, the title compound was prepared following a procedure similar to that described for step 2 of intermediate B2, and obtained as a pale yellow solid (0.052 g, 63% yield), which was not further purified. LCMS: 266.0 [M+H]

[0270] [ka] A mixture of 1-(3-(benzyloxy)cyclobutyl)-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A17, 0.400 g, 0.712 mmol), 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.186 g, 0.783 mmol), and K2CO3 (0.197 g, 1.424 mmol) in dioxane (7 mL), ethanol (3 mL), and water (3 mL) was purged with N2 for 10 minutes. Then PdCl2 (dppf) (0.026 g, 0.036 mmol) was added, and the resulting mixture was again purged with N2 for 10 minutes, and then stirred in a sealed tube at 80°C for 16 hours. After the reaction was complete (shown by TLC and LC-MS), the reaction mixture was filtered through a diatomaceous earth pad and then rinsed with siRNA (3 × 10 mL × 3). The combined filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by flash chromatography (silica gel, 230-400 mesh, in DCM, eluted with 4% MeOH) to obtain the title compound as a light brown solid (0.179 g, yield 62%). 1 H NMR (400 MHz, DMSO-d6) δ = 8.21 (s, 1H), 7.20-7.40 (m, 7H), 6.89-6.93 (m, 1H), 5.42-5.48 (m, 3H), 4.47-4.48 (m, 3H), 2.74-2.83 (m, 2H), 2.60 (s, 2H).LCMS: 405.1 [M+H]

[0271] [ka] Starting with 1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A1, 0.250 g, 0.830 mmol) and 2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.213 g, 0.913 mmol), the title compound was prepared following the same procedure as described for intermediate B26, obtained as a brown gum (0.22 g), and the process proceeded without further purification. LCMS: 281.3 [M+H]

[0272] [ka] Starting with 3-iodo-1-(1-methylazetidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A18, 0.380 g, 1.151 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.300 g, 1.266 mmol), the title compound was obtained as a brown solid (0.26 g, 48% yield) following a procedure similar to that described for intermediate B26. LCMS: 314.4 [M+H]

[0273] [ka] Starting with 1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A1, 0.165 g, 0.548 mmol) and 2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (prepared as described in PCT publication number WO2020 / 172093, 0.300 g, 1.266 mmol), the title compound was obtained as a pale yellow solid (0.051 g, 27% yield) by following a procedure similar to that described for intermediate B26. LCMS: 303.2 [M+H]

[0274] [ka] Starting with 1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A1, 0.250 g, 0.830 mmol) and (2-amino-5-(4,4,5,5-tetramethyl-1,3-dioxolan-2-yl)phenyl)methanol (prepared as described in PCT publication number WO2011 / 8130628, 0.207 g, 0.830 mmol), the title compound was obtained as a pale pink solid (0.122 g, 47% yield) by following a procedure similar to that described for intermediate B26. LCMS: 297.1 [M+H]

[0275] [ka] Starting with 1-cyclopropyl-N-(2,4-dimethoxybenzyl)-3-iodo-1H-pyrazolo[4,3-c]pyridine-4-amine (A19, 0.300 g, 0.666 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-amine (0.147 g, 0.666 mmol), the title compound was obtained as a brown gum (0.11 g, 30% yield) following a procedure similar to that described for intermediate B26. LCMS: 416.9 [M+H]

[0276] [ka] Starting with 2-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-1-yl)ethane-1-ol (A20, 0.420 g, 1.377 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.327 g, 1.379 mmol), the title compound was obtained as a brown solid (0.390 g, 92% yield) by following a procedure similar to that described for intermediate B26. LCMS: 289.1 [M+H]

[0277] [ka] Starting with 3-iodo-1-(2-methoxyethyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A21, 0.620 g, 1.943 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.461 g, 1.943 mmol), the title compound was obtained as a brown solid (0.42 g) following a procedure similar to that described for intermediate B26, and was used without further purification. LCMS: 302.9 [M+H]

[0278] [ka] Starting with 1-cyclopropyl-N-(2,4-dimethoxybenzyl)-3-iodo-1H-pyrazolo[4,3-c]pyridine-4-amine (A19, 0.600 g, 1.333 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.316 g, 1.333 mmol), the title compound was obtained as a brown solid (0.43 g, 72% yield) by following a procedure similar to that described for intermediate B26. 1 H NMR (400 MHz, DMSO-d6) δ = 7.82 (d, J = 6.0 Hz, 1H), 7.07-7.19 (m, 3H), 6.83-6.89 (m, 2H), 6.51 (d, J = 2.0 Hz, 1H), 6.40-6.43 (m, 1H), 5.62 (t, J = 5.6 Hz, 1H), 5.49 (bs, 2H), 4.49 (d, J = 5.6 Hz, 2H), 3.65-3.72 (m, 7H), 1.08-1.09 (m, 4H).LCMS: 434.2 [M+H]

[0279] [ka] Starting with 1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A1, 0.250 g, 0.830 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-amine (0.183 g, 0.830 mmol), the title compound was obtained as a brown solid (0.085 g, 38% yield) following a procedure similar to that described for intermediate B26. LCMS: 268.6 [M+H]

[0280] [ka] Starting with 3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A22, 0.500 g, 1.278 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.454 g, 1.917 mmol), the title compound was obtained as a brown solid (0.16 g, 30% yield) by following a procedure similar to that described for intermediate B26. 1 H NMR (400 MHz, DMSO-d6) δ = 8.26 (s, 1H), 7.20-7.27 (m, 2H), 6.88-6.93 (m, 1H), 5.62 (s, 2H), 5.52 (bs, 2H), 3.62 (t, J = 8.0 Hz, 2H), 0.84 (t, J = 8.0 Hz, 2H), -0.12 (s, 9H).LCMS: 375.4 [M+H]

[0281] [ka] Starting with 3-iodo-1-(1-methylpiperidine-4-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A23, 0.180 g, 0.503 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.131 g, 0.553 mmol), the title compound was obtained as a brown solid (0.22 g, 72% yield) following a procedure similar to that described for intermediate B26. LCMS: 342.2 [M+H]

[0282] General procedure for synthesizing carbamate intermediate C Pyridine (1.2 eq) and phenyl chloroformate (1.5 eq) were added to a solution of amine (1.0 eq) in THF (10 vol) (0°C). The reaction mixture was allowed to warm to 25°C and stirred for 12 hours. After the reaction was complete (shown by TLC), the mixture was diluted with HCl (10 mL) and washed with brine (5 mL). The organic layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude product, which was purified by flash chromatography (silica gel 230-400 mesh, in petroleum ether, eluted with 10-20% HCl) to obtain the target carbamate.

[0283] The following carbamates were prepared using the general procedure described above. [Table 26] [Table 27] [Table 28] Note: The amines used in the synthesis of carbamates are either commercially available or synthesized using procedures described in the literature, as follows:

[0284] 3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-amine (precursor of C6) and 5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-amine (precursor of C7) were synthesized from 1-(trifluoromethyl)cyclopropane-1-carboxylic acid methyl ester, as described in Synthesis 2013, 45, 171-173.

[0285] 3-(1,1,1-trifluoro-2-methylpropan-2-yl)isoxazole-5-amine (precursor of C8) and 3-(2-fluoropropan-2-yl)isoxazole-5-amine (precursor of C9) were synthesized from methyl 3,3,3-trifluoro-2,2-dimethylpropanoate and methyl 2-fluoro-2-methylpropionate, respectively, according to the procedure described in Synthesis 2013, 45, 171-173.

[0286] 2-(5-aminoisoxazole-3-yl)-2-methylpropanenitrile (a precursor of C20) was synthesized from 2,2-dimethyl-3-oxopentanedinitrile, as described in J. Med. Chem. 2012, 55, 1082~1105.

[0287] 3-(((Tert-butyldimethylsilyl)oxy)methyl)isoxazole-5-amine (precursor of C21) was synthesized from 4-((tert-butyldiphenylsilyl)oxy)-3-oxobutanenitrile, as described in PCT publication number WO2013 / 104561.

[0288] 3-(Tert-butyl)-4-methylisoxazole-5-amine (a precursor of C22) was synthesized from 2,4,4-trimethyl-3-oxopentannitrile, as described in PCT publication number WO2012 / 019015.

[0289] [ka] NH2OH·H2SO4 (0.699 g, 4.25 mmol) was added to a stirred solution of 3-(1-methylcyclobutyl)-3-oxopropanenitrile (prepared as described in PCT publication number WO2017 / 060874, 0.500 g, 3.86 mmol) and sodium hydroxide (0.170 g, 4.25 mmol) in EtOH (10 mL) and water (10 mL). The pH of the resulting mixture was adjusted to 7.5 using aqueous NaOH solution (1 M), and the reaction mixture was stirred at 80°C for 15 hours. After the reaction was complete (shown by TLC), the reaction mixture was concentrated under reduced pressure to obtain the residue, which was placed in DCM (25 mL), washed with water (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude material was purified by flash chromatography (silica gel, 230-400 mesh, in petroleum ether, eluted with 40% ethyl acetate) to obtain the title product as an off-white solid (0.110 g, yield 19%). 1 H NMR (400 MHz, CDCl3) δ = 5.04 (s, 1H), 2.43-2.49 (m, 2H), 1.96-2.02 (m, 4H), 1.50 (s, 3H).LCMS: 153.2 [M+H]

[0290] Preparation of Examples General urea formation procedure for the synthesis of Examples 1-72

[0291] Method A: Triethylamine (2.0 eq.) was added to a mixture of amine intermediate D (1.0 eq.) and carbamate intermediate E (1.0 eq.) in THF (10 Vol.). The resulting mixture was stirred in a sealed tube at 60°C for 12 hours. After the reaction was complete (shown by LC-MS), the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by reverse-phase preparative HPLC to obtain the target product.

[0292] Method B-DMAP (0.05 eq.) and DIPEA (1.5 eq.) were added to a solution of amine intermediate D (1.0 eq.) and carbamate intermediate E (1.0 eq.) in THF (10 Vol.), and the resulting mixture was stirred in a sealed tube at 60°C for 12 hours. After the reaction was complete (shown by LC-MS), the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by reverse-phase preparative HPLC to obtain the target product.

[0293] The following compounds were prepared using the general procedure described above.

[0294] [ka] Starting from 3-(4-aminophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B2, 0.250 g, 0.93 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.242 g, 0.93 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.150 g, 37% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 10.15 (bs, 1H), 9.09 (bs, 1H), 8.24 (bs, 1H), 7.55-7.66 (m, 4H), 6.10 (s, 1H), 5.03-5.10 (m, 1H), 1.49 (d, J = 6.4 Hz, 6H), 1.27 (s, 9H).LCMS: 435.3 [M+H]

[0295] [ka] Starting from 3-(4-aminophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B4, 0.210 g, 0.744 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.193 g, 0.744 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.053 g, yield 16%). 1 H NMR (400 MHz, DMSO-d6) δ = 10.23 (bs, 1H), 9.15 (bs, 1H), 8.31 (s, 1H), 7.74 (s, 4H), 6.16 (s, 1H), 6.06-6.10 (m, 1H), 5.16 (t, J = 6.4 Hz, 2H), 5.05 (t, J = 6.8 Hz, 2H), 1.33 (s, 9H).LCMS: 449.2[M+H]

[0296] [ka] Starting from 3-(4-aminophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B4, 0.070 g, 0.248 mmol) and phenyl (5-(tert-butyl)isoxazole-3-yl)carbamate (C5, 0.065 g, 0.248 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.035 g, 32% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 9.59 (bs, 1H), 9.07 (bs, 1H), 8.26 (s, 1H), 7.66 (bs, 4H), 6.53 (s, 1H), 5.98-6.04 (m, 1H), 5.09 (t, J = 6.4 Hz, 2H), 5.00 (t, J = 6.8 Hz, 2H), 1.31 (s, 9H).LCMS: 449.2[M+H]

[0297] [ka] Starting from 3-(4-aminophenyl)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B12, 0.05 g, 0.16 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.042 g, 0.16 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.013 g, 20% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 10.13 (bs, 1H), 9.08 (bs, 1H), 8.25 (s, 1H), 7.61-7.67 (m, 4H), 6.10 (s, 1H), 4.90-4.98 (m, 1H), 4.00-4.03 (m, 2H), 3.53-3.58 (m, 2H), 2.19-2.26 (m, 2H), 1.87-1.91 (m, 2H), 1.27 (s, 9H).LCMS: 477.1[M+H]

[0298] [ka] Starting from 3-(4-aminophenyl)-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B10, 0.083 g, 0.28 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.072 g, 0.28 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.048 g, 36% yield). 1H NMR (400 MHz, DMSO-d6) δ = 10.17 (bs, 1H), 9.08 (bs, 1H), 8.26 (s, 1H), 7.60-7.67 (m, 4H), 6.10 (s, 1H), 5.49-5.52 (m, 1H), 4.07-4.14 (m, 2H), 3.89-3.97 (m, 2H), 2.38-2.43 (m, 2H), 1.27 (s, 9H).LCMS: 463.0[M+H]

[0299] [ka] Starting from 3-(4-amino-3-chlorophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B21, 0.080 g, 0.264 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.067 g, 0.264 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.003 g, yield 3%). 1 H NMR (400 MHz, DMSO-6) δ = 10.85 (bs, 1H), 8.66 (bs, 1H), 8.35 (d, J = 8.4 Hz, 1H), 8.25 (s, 1H), 7.72 (d, J = 2.0 Hz, 1H), 7.61-7.64 (m, 1H), 6.12 (s, 1H), 5.04-5.10 (m, 1H), 1.50 (d, J = 6.8 Hz, 6H), 1.28 (s, 9H).LCMS: 469.1[M+H]

[0300] [ka] Starting from 3-(4-aminophenyl)-1-cyclobutyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B8, 0.140 g, 0.49 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.130 g, 0.49 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.067 g, 29% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 10.15 (bs, 1H), 9.09 (bs, 1H), 8.24 (s, 1H), 7.62-7.68 (m, 4H), 6.10 (s, 1H), 5.32-5.36 (m, 1H), 2.69-2.74 (m, 2H), 2.39-2.42 (m, 2H), 1.87-1.89 (m, 2H), 1.27 (s, 9H).LCMS: 447.2[M+H]

[0301] [ka] Starting from 3-(4-aminophenyl)-1-cyclobutyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B9, 0.100 g, 0.33 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.087 g, 0.33 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as a pale yellow solid (0.005 g, yield 4%). 1 H NMR (400 MHz, DMSO-d6) δ = 9.10 (bs, 1H), 8.23-8.28 (m, 2H), 7.47-7.53 (m, 2H), 6.90 (bs, 1H), 6.09 (s, 1H), 5.29-5.37 (m, 1H), 2.67-2.73 (m, 2H), 2.38-2.41 (m, 2H), 1.85-1.89 (m, 2H), 1.26 (s, 9H).LCMS: 465.1[M+H]

[0302] [ka] Starting from 1-(4-amino-3-(4-aminophenyl)-1H-pyrazolo[3,4-d]pyrimidine-1-yl)-2-methylpropan-2-ol (B18, 0.119 g, 0.4 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.103 g, 0.4 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.017 g, 9% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 9.44 (bs, 1H), 8.24 (s, 1H), 7.59-7.68 (m, 4H), 6.07 (s, 1H), 4.81 (bs, 1H), 4.27 (bs, 2H), 1.26 (s, 9H), 1.15 (s, 6H).LCMS: 465.1[M+H]

[0303] [ka] Starting from 1-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-d]pyrimidine-1-yl)-2-methylpropan-2-ol (B19, 0.077 g, 0.243 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.063 g, 0.243 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.013 g, yield 11%). 1 H NMR (400 MHz, CD3OD) δ = 8.33-8.37 (m, 1H), 8.29 (s, 1H), 7.53-7.59 (m, 2H), 6.19 (s, 1H), 4.42 (s, 2H), 1.35 (s, 9H), 1.29 (s, 6H).LCMS: 483.3[M+H]

[0304] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B3, 0.200 g, 0.69 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.182 g, 0.69 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.011 g, yield 3%). 1 H NMR (400 MHz, DMSO-d6) δ = 10.40 (bs, 1H), 8.93 (bs, 1H), 8.24-8.30 (m, 2H), 7.47-7.52 (m, 2H), 6.95 (bs, 2H), 6.11 (s, 1H), 5.03-5.10 (m, 1H), 1.49 (d, J = 6.4 Hz, 6H), 1.27 (s, 9H).LCMS: 453.2[M+H]

[0305] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B5, 0.200 g, 0.66 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.173 g, 0.66 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.027 g, yield 10%). 1 H NMR (400 MHz, DMSO-d6) δ = 8.97 (bs, 1H), 8.29-8.33 (m, 1H), 8.25 (s, 1H), 7.52-7.59 (m, 2H), 6.11 (s, 1H), 5.98-6.05 (m, 1H), 5.01-5.11 (m, 2H), 4.98-4.99 (m, 2H), 1.27 (s, 9H).LCMS: 467.1[M+H]

[0306] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.150 g, 0.52 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.137 g, 0.52 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as a white solid (0.035 g, yield 15%). 1 H NMR (400 MHz, DMSO-d6) δ = 10.42 (bs, 1H), 8.98 (bs, 1H), 8.25-8.29 (m, 2H), 7.44-7.50 (m, 2H), 6.95 (bs, 2H), 6.10 (s, 1H), 3.85-3.87 (m, 1H), 1.27 (s, 9H), 1.20-1.23 (m, 2H), 1.08-1.10 (m, 2H).LCMS: 451.2[M+H]

[0307] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(1-(oxetan-3-yl)piperidine-4-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B16, 0.050 g, 0.13 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.034 g, 0.13 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.006 g, yield 8%). 1H NMR (400 MHz, CD3OD) δ = 8.32-8.36 (m, 1H), 8.27 (s, 1H), 7.51-7.57 (m, 2H), 6.19 (s, 1H), 4.65-4.82 (m, 5H), 3.61-3.64 (m, 1H), 2.98-3.01 (m, 2H), 2.37-2.44 (m, 2H), 2.13-2.19 (m, 2H), 2.04-2.07 (m, 2H), 1.35 (s, 9H).LCMS: 550.0[M+H]

[0308] [ka] Starting from 4-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-d]pyrimidine-1-yl)cyclohexane-1-ol (B15, 0.075 g, 0.22 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.057 g, 0.22 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.007 g, yield 6%). 1 H NMR (400 MHz, CD3OD) δ = 8.32-8.36 (m, 1H), 8.27 (s, 1H), 7.50-7.55 (m, 2H), 6.19 (s, 1H), 4.74-4.80 (m, 1H), 3.72-3.73 (m, 1H), 2.14-2.21 (m, 4H), 2.03-2.06 (m, 2H), 1.55-1.59 (m, 2H), 1.35 (s, 9H).LCMS: 509.2[M+H]

[0309] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(3,3-difluorocyclobutyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B14, 0.110 g, 0.33 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.086 g, 0.33 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.020 g, yield 13%). 1 H NMR (400 MHz, DMSO-d6) δ = 9.28 (bs, 1H), 8.26-8.30 (m, 2H), 7.48-7.55 (m, 2H), 7.08 (bs, 1H), 6.07 (s, 1H), 5.28-5.33 (m, 1H), 3.25-3.34 (m, 4H), 1.27 (s, 9H).LCMS: 501.1[M+H]

[0310] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(pyridine-4-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B20, 0.140 g, 0.43 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.113 g, 0.43 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.006 g, yield 3%). 1 H NMR (400 MHz, DMSO-d6) δ = 10.44 (bs, 1H), 9.00 (bs, 1H), 8.73-8.74 (m, 2H), 8.34-8.47 (m, 4H), 7.59-7.68 (m, 2H), 6.13 (s, 1H), 1.28 (s, 9H).LCMS: 488.2[M+H]

[0311] [ka] Starting from 3-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-d]pyrimidine-1-yl)tetrahydrothiophene 1,1-dioxide (B17, 0.030 g, 0.082 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.021 g, 0.082 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as a white solid (0.003 g, yield 7%). 1 H NMR (400 MHz, CD3OD) δ = 8.31-8.36 (m, 2H), 7.54-7.60 (m, 2H), 6.19 (s, 1H), 5.78-5.80 (m, 1H), 3.58-3.63 (m, 3H), 2.78-2.89 (m, 2H), 1.32 (s, 9H).LCMS: 529.2[M+H]

[0312] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(tetrahydro-2H-pyran-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B13, 0.015 g, 0.045 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.011 g, 0.045 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.002 g, 9% yield). 1 H NMR (400 MHz, CD3OD) δ = 8.54 (s, 1H), 8.35 (d, J = 7.2 Hz, 1H), 8.28 (s, 1H), 7.50-7.56 (m, 2H), 6.19 (s, 1H), 4.60 (s, 1H), 3.87-4.05 (m, 3H), 3.50-3.58 (m, 1H), 2.38-2.42 (m, 1H), 2.21-2.23 (m, 1H), 1.92-2.03 (m, 2H), 1.35 (s, 9H).LCMS: 495.1[M+H]

[0313] [ka] Starting from 3-(4-aminophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B7, 0.100 g, 0.38 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.100 g, 0.38 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as a white solid (0.045 g, yield 27%). 1 H NMR (400 MHz, DMSO-d6) δ = 10.11 (bs, 1H), 9.10 (bs, 1H), 8.26 (s, 1H), 7.58-7.65 (m, 4H), 6.09 (s, 1H), 3.83-3.89 (m, 1H), 1.24 (s, 9H), 1.19-1.23 (m, 2H), 1.06-1.11 (m, 2H).LCMS: 433.2[M+H]

[0314] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.100 g, 0.31 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.091 g, 0.31 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.020 g, yield 12%). 1H NMR (400 MHz, DMSO-d6) δ = 10.63 (bs, 1H), 8.92 (bs, 1H), 8.25-8.29 (m, 2H), 7.44-7.51 (m, 2H), 6.87 (bs, 2H), 6.20 (s, 1H), 3.84-3.89 (m, 1H), 1.45-1.76 (m, 2H), 1.36-1.41 (m, 2H), 1.19-1.23 (m, 2H), 1.06-1.11 (m, 2H).LCMS: 503.1[M+H]

[0315] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.050 g, 0.17 mmol) and phenyl (3-(1,1,1-trifluoro-2-methylpropan-2-yl)isoxazole-5-yl)carbamate (C8, 0.045 g, 0.17 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.004 g, yield 4%). 1 H NMR (400 MHz, DMSO-d6) δ = 10.62 (bs, 1H), 8.96 (bs, 1H), 8.25-8.28 (m, 2H), 7.45-7.52 (m, 2H), 6.99 (bs, 2H), 6.25 (s, 1H), 3.84-3.90 (m, 1H), 1.53 (s, 6H), 1.19-1.23 (m, 2H), 1.10-1.12 (m, 2H).LCMS: 505.1[M+H].

[0316] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B5, 0.150 g, 0.5 mmol) and phenyl (5-(tert-butyl)isoxazole-3-yl)carbamate (C5, 0.130 g, 0.5 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.015 g, yield 6%). 1 H NMR (400 MHz, DMSO-d6) δ = 10.00 (bs, 1H), 9.09 (bs, 1H), 8.32-8.37 (m, 1H), 8.26 (s, 1H), 7.51-7.59 (m, 2H), 6.80 (bs, 2H), 6.53 (s, 1H), 5.99-6.04 (m, 1H), 5.10 (t, J = 6.4 Hz, 2H), 5.00 (t, J = 6.4 Hz, 2H), 1.31 (s, 9H).LCMS: 467.2[M+H]

[0317] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B11, 0.200 g, 0.59 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.154 g, 0.59 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as a white solid (0.030 g, yield 10%). 1H NMR (400 MHz, DMSO-d6) δ = 10.40 (bs, 1H), 8.92 (bs, 1H), 8.26-8.31 (m, 2H), 7.47-7.54 (m, 2H), 6.80 (bs, 2H), 6.11 (s, 1H), 5.47-5.53 (m, 1H), 4.05-4.13 (m, 2H), 3.87-3.96 (m, 2H), 2.40-2.41 (m, 2H), 1.27 (s, 9H).LCMS: 481.2[M+H]

[0318] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.070 g, 0.24 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.077 g, 0.24 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.013 mg, yield 11%). 1 H NMR (400 MHz, DMSO-d6) δ = 11.78 (bs, 1H), 8.76 (s, 1H), 7.43-7.45 (m, 2H), 6.84-6.88 (m, 2H), 5.46 (s, 2H), 3.91-3.97 (m, 1H), 1.49-1.54 (m, 4H), 1.19-1.22 (m, 2H), 1.10-1.15 (m, 2H).LCMS: 503.1[M+H]

[0319] [ka] Starting from 1-allyl-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B6, 0.130 g, 0.45 mmol) and phenyl (5-(tert-butyl)isoxazole-3-yl)carbamate (C5, 0.119 g, 0.45 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.012 g, 6% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 10.46 (bs, 1H), 9.52 (bs, 1H), 8.24-8.28 (m, 2H), 7.45-7.50 (m, 2H), 6.95 (bs, 2H), 6.52 (s, 1H), 6.00-6.10 (m, 1H), 5.18-5.21 (m, 2H), 5.13 (d, J = 1.6 Hz, 2H), 1.31 (s, 9H).LCMS: 451.2[M+H]

[0320] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.050 g, 0.176 mmol) and phenyl (3-(2-fluoropropan-2-yl)isoxazole-5-yl)carbamate (C9, 0.047 g, 0.176 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.007 mg, 9% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 8.32-8.35 (m, 1H), 8.25 (s, 1H), 7.32-7.37 (m, 2H), 5.92 (s, 1H), 3.83-3.89 (m, 1H), 1.66 (s, 3H), 1.61 (s, 3H), 1.20-1.24 (m, 2H), 1.07-1.09 (m, 2H).LCMS: 455.1[M+H]

[0321] [ka] Starting from 3-(4-amino-3-chlorophenyl)-1-methyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B22, 0.068 g, 0.248 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.064 g, 0.248 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.005 g, 4% yield). 1 H NMR (400 MHz, CD3OD) δ = 8.70-8.68 (m, 1H), 7.47-7.67 (m, 2H), 7.00-7.02 (m, 1H), 6.20 (s, 1H), 4.10 (s, 3H), 1.34 (s, 9H).LCMS: 441.1[M+H]

[0322] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-methyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B23, 0.052 g, 0.201 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.052 g, 0.201 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.005 g, 6% yield). 1 H NMR (400 MHz, CD3OD) δ = 10.60 (bs, 1H), 9.05 (bs, 1H), 8.25-8.30 (m, 2H), 7.45-7.51 (m, 2H), 6.90 (s, 2H), 6.09 (s, 1H), 3.95 (s, 3H), 1.27 (s, 9H).LCMS: 423.1[MH]

[0323] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[4,3-c]pyridine-4-amine (B24, 0.070 g, 0.24 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.064 g, 0.24 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as an off-white solid (0.004 g, 4% yield). 1 H NMR (400 MHz, DMSO-d6) δ = 10.52 (bs, 1H), 9.09 (bs, 1H), 8.23-8.27 (m, 1H), 7.80 (s, 1H), 7.44-7.53 (m, 2H), 6.90 (d, J = 6.0 Hz, 1H), 6.10 (s, 1H), 5.86 (bs, 2H), 3.72-3.74 (m, 1H), 1.27 (s, 9H), 1.12-1.13 (m, 4H).LCMS: 450.2[M+H]

[0324] [ka] Starting from 3-(4-aminophenyl)-1-cyclopropyl-1H-pyrazolo[4,3-c]pyridine-4-amine (B25, 0.052 g, 0.2 mmol) and phenyl (3-(tert-butyl)isoxazole-5-yl)carbamate (C4, 0.05 g, 0.2 mmol), the title compound was prepared according to the general procedure for urea formation (Method A) and obtained as a white solid (0.007 g, yield 8%). 1 H NMR (400 MHz, DMSO-d6) δ = 10.85 (bs, 1H), 9.68 (bs, 1H), 7.78 (bs, 1H), 7.66 (d, J = 8.4 Hz, 2H), 7.56 (d, J = 8.4 Hz, 2H), 6.90 (d, J = 6.0 Hz, 1H), 6.08 (s, 1H), 5.75 (bs, 2H), 3.68-3.73 (m, 1H), 1.26 (s, 9H), 1.11-1.12 (m, 4H).LCMS: 432.3[M+H].

[0325] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.050 g, 0.176 mmol) and phenyl (3-(1-methylcyclopropyl)isoxazole-5-yl)carbamate (C10, 0.045 g, 0.176 mmol), the title compound was obtained as an off-white solid (0.009 g, yield 11%) according to the general procedure for urea formation (Method A). 1 H NMR (400 MHz, DMSO-d6) δ = 10.29 (bs, 1H), 8.90 (bs, 1H), 8.24-8.28 (m, 2H), 7.44-7.51 (m, 2H), 6.94 (bs, 2H), 5.86 (s, 1H), 3.84-3.89 (m, 1H), 1.38 (s, 3H), 1.19-1.22 (m, 2H), 1.08-1.11 (m, 2H), 0.94-0.97 (m, 2H), 0.82-0.85 (m, 2H).LCMS: 449.1 [M+H]

[0326] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.050 g, 0.176 mmol) and phenyl (5-(tert-butyl)-1,3,4-thiadiazole-2-yl)carbamate (C11, 0.049 g, 0.176 mmol), the title compound was obtained as an off-white solid (0.012 mg, yield 14%) according to the general procedure for urea formation (Method A). 1H NMR (400 MHz, DMSO-d6) δ = 8.67 (bs, 1H), 8.33-8.41 (m, 1H), 8.25 (s, 1H), 7.34-7.39 (m, 2H), 6.87 (bs, 2H), 3.83-3.87 (m, 1H), 1.34 (s, 9H), 1.20-1.23 (m, 2H), 1.06-1.11 (m, 2H).LCMS: 468.2 [M+H]

[0327] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.050 g, 0.176 mmol) and phenyl (4-(tert-butyl)thiazole-2-yl)carbamate (C12, 0.049 g, 0.176 mmol), the title compound was obtained as an off-white solid (0.004 g, 5% yield) according to the general procedure for urea formation (Method A). 1 H NMR (400 MHz, CD3OD) δ = 8.45-8.49 (m, 1H), 8.40 (s, 1H), 7.50-7.58 (m, 2H), 6.65 (s, 1H), 3.96-4.02 (m, 1H), 1.31-1.40 (m, 11H), 1.21-1.25 (m, 2H).LCMS: 467.2 [M+H]

[0328] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.050 g, 0.176 mmol) and phenyl (3-(tert-butyl)isothiazol-5-yl)carbamate (C13, 0.049 g, 0.176 mmol), the title compound was obtained as an off-white solid (0.016 g, 19% yield) according to the general procedure for urea formation (Method A). 1H NMR (400 MHz, DMSO-d6) δ = 11.42 (bs, 1H), 8.51 (bs, 1H), 8.26 (s, 1H), 8.07-8.11 (m, 1H), 7.39-7.43 (m, 2H), 6.70 (s, 1H), 3.84-3.87 (m, 1H), 1.21-1.26 (m, 11H), 1.08-1.10 (m, 2H).LCMS: 465.1 [MH]

[0329] [ka] Starting with phenyl (4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-3-yl)-2-fluorophenyl)carbamate (C3, 0.050 g, 0.124 mmol) and 5-(tert-butyl)-1,3,4-oxadiazole-2-amine (0.017 g, 0.124 mmol), the title compound was obtained as an off-white solid (0.002 g, 4% yield) according to the general procedure for urea formation (Method A). 1 H NMR (400 MHz, CD3OD) δ = 8.37-8.43 (m, 2H), 7.46-7.62 (m, 2H), 3.93-3.95 (m, 1H), 1.45 (s, 9H), 1.31-1.38 (m, 2H), 1.20-1.23 (m, 2H).LCMS: 451.9 [M+H]

[0330] [ka] Starting from phenyl (4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-3-yl)-2-fluorophenyl)carbamate (C3, 0.050 g, 0.124 mmol) and 3-(tert-butyl)-1,2,4-thiadiazole-5-amine (0.019 g, 0.124 mmol), the title compound was obtained as an off-white solid (0.001 g, yield 1%) according to the general procedure for urea formation (Method A). 1H NMR (400 MHz, CD3OD) δ = 8.27-8.39 (m, 2H), 7.51-7.56 (m, 2H), 3.80-3.86 (m, 1H), 1.40 (s, 9H), 1.31-1.33 (m, 2H), 1.17-1.21 (m, 2H).LCMS: 468.0 [M+H]

[0331] [ka] Starting with phenyl (4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-3-yl)-2-fluorophenyl)carbamate (C3, 0.050 g, 0.124 mmol) and 1-(tert-butyl)-1H-1,2,4-triazole-3-amine (0.017 g, 0.124 mmol), the title compound was obtained as an off-white solid (0.009 g, 15% yield) according to the general procedure for urea formation (Method A). 1 H NMR (400 MHz, DMSO-d6) δ = 10.80 (bs, 1H), 10.21 (bs, 1H), 8.55 (s, 1H), 8.42-8.46 (m, 1H), 8.26 (s, 1H), 7.44-7.52 (m, 2H), 6.94 (bs, 2H), 3.83-3.88 (m, 1H), 1.57 (s, 9H), 1.19-1.23 (m, 2H), 1.06-1.11 (m, 2H).LCMS: 451.0 [M+H]

[0332] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.050 g, 0.176 mmol) and phenyl (3-(trifluoromethyl)isoxazole-5-yl)carbamate (C14, 0.048 g, 0.176 mmol), the title compound was obtained as a white solid (0.004 g, 5% yield) according to the general procedure for urea formation (Method A).1 H NMR (400 MHz, DMSO-d6) δ = 11.11 (bs, 1H), 8.97 (bs, 1H), 8.23-8.27 (m, 2H), 7.46-7.52 (m, 2H), 6.95 (bs, 2H), 6.53 (s, 1H), 3.84-3.88 (m, 1H), 1.19-1.24 (m, 2H), 1.07-1.12 (m, 2H).LCMS: 462.9 [M+H]

[0333] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.05 g, 0.176 mmol) and phenyl (3-(pentan-3-yl)isoxazole-5-yl)carbamate (C15, 0.048 g, 0.176 mmol), the title compound was obtained as a light brown solid (0.006 g, yield 7%) according to the general procedure for urea formation (Method A). 1 H NMR (400 MHz, DMSO-d6) δ = 10.44 (bs, 1H), 8.97 (bs, 1H), 8.26-8.29 (m, 2H), 7.44-7.50 (m, 2H), 6.81 (bs, 2H), 6.00 (s, 1H), 3.84-3.86 (m, 1H), 1.51-1.66 (m, 4H), 1.08-1.21 (m, 4H), 0.79-0.82 (m, 6H).LCMS: 465.0 [M+H]

[0334] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.050 g, 0.176 mmol) and phenyl (3-isopropylisoxazole-5-yl)carbamate (C16, 0.043 g, 0.176 mmol), the title compound was obtained as an off-white solid (0.007 g, 9% yield) according to the general procedure for urea formation (Method A). 1 H NMR (400 MHz, DMSO-d6) δ = 10.49 (bs, 1H), 9.01 (bs, 1H), 8.27-8.32 (m, 2H), 7.44-7.52 (m, 2H), 6.07 (s, 1H), 3.87-3.92 (m, 1H), 2.90-2.97 (m, 1H), 1.18-1.24 (m, 8H), 1.10-1.12 (m, 2H).LCMS: 437.0 [M+H]

[0335] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.050 g, 0.176 mmol) and phenyl (3-ethylisoxazole-5-yl)carbamate (C17, 0.041 g, 0.176 mmol), the title compound was obtained as an off-white solid (0.006 g, yield 8%) according to the general procedure for urea formation (Method A). 1 H NMR (400 MHz, DMSO-d6) δ = 10.44 (bs, 1H), 8.94 (bs, 1H), 8.26-8.30 (m, 2H), 7.44-7.51 (m, 2H), 6.99 (bs, 2H), 6.05 (s, 1H), 3.84-3.89 (m, 1H), 2.53-2.59 (m, 2H), 1.18-1.21 (m, 5H), 1.08-1.10 (m, 2H).LCMS: 423.0 [M+H]

[0336] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.080 g, 0.281 mmol) and phenyl (3-(sec-butyl)isoxazole-5-yl)carbamate (C18, 0.073 g, 0.281 mmol), the title compound was obtained as a white solid (0.004 g, 4% yield) according to the general procedure for urea formation (Method A). 1 H NMR (400 MHz, DMSO-d6) δ = 10.42 (bs, 1H), 8.96 (bs, 1H), 8.26-8.33 (m, 2H), 7.44-7.52 (m, 2H), 6.04 (s, 1H), 3.87-3.92 (m, 1H), 2.68-2.73 (m, 1H), 1.54-1.60 (m, 2H), 1.08-1.24 (m, 7H), 0.82-0.84 (m, 3H).LCMS: 451.1 [M+H]

[0337] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.100 g, 0.352 mmol) and phenyl (3-(1-methylcyclobutyl)isoxazole-5-yl)carbamate (C19, 0.096 g, 0.352 mmol), the title compound was obtained as a white solid (0.002 g, yield 1%) according to the general procedure for urea formation (Method A). 1 H NMR (400 MHz, CD3OD) δ = 8.35-8.41 (m, 2H), 7.49-7.56 (m, 2H), 6.16 (s, 1H), 3.98-4.02 (m, 1H), 2.46-2.51 (m, 2H), 1.96-2.13 (m, 4H), 1.54 (s, 3H), 1.23-1.38 (m, 4H).LCMS: 463.1 [M+H]

[0338] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[4,3-c]pyridine-4-amine (B24, 0.160 g, 0.565 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.176 g, 0.565 mmol), the title compound was obtained as an off-white solid (0.012 g, 4% yield) according to the general procedure for urea formation (Method A). 1 H NMR (400 MHz, DMSO-d6) δ = 10.69 (bs, 1H), 8.97 (bs, 1H), 8.23-8.27 (m, 1H), 7.80 (d, J = 6.0 Hz, 1H), 7.46-7.54 (m, 2H), 6.91 (d, J = 6.0 Hz, 1H), 6.22 (s, 1H), 5.85 (bs, 2H), 3.72-3.74 (m, 1H), 1.39-1.49 (m, 4H), 1.12-1.13 (m, 4H).LCMS: 502.2 [M+H]

[0339] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-(3-(benzyloxy)cyclobutyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B26, 0.090 g, 0.223 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.076 g, 0.245 mmol), the title compound was obtained as a brown gum (70 mg) according to the general procedure for urea formation (Method B), and used without further purification. LCMS: 623.4 [M+H]

[0340] [ka] Boron trichloride (1 M, 0.899 mL, 0.899 mmol in DCM) was added dropwise to a solution of 1-(4-(4-amino-1-(3-(benzyloxy)cyclobutyl)-1H-pyrazolo[3,4-d]pyrimidine-3-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)urea (0.070 g, 0.112 mmol) in 5 mL of dry DCM (-60°C), and the resulting mixture was stirred at 0°C for 3 hours. After the reaction was complete (shown by TLC and LCMS), the reaction mixture was cooled to -70°C, neutralized with aqueous ammonia (25% aqueous solution), and extracted with DCM (10 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by preparative HPLC (eluting with 1% TFA in a mixture of water and ACN) to obtain the title product as a white solid (0.009 g, yield 15%). 1 H NMR (400 MHz, CD3OD) δ = 8.34-8.38 (m, 2H), 7.53-7.61 (m, 2H), 6.34 (s, 1H), 5.61-5.64 (m, 1H), 4.73-4.79 (m, 1H), 2.94-3.01 (m, 2H), 2.57-2.63 (m, 2H), 1.39-1.48 (m, 4H).LCMS: 533.7 [M+H]

[0341] [ka] Starting with 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-amine (0.100 g, 0.454 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.142 g, 0.454 mmol), the title compound was obtained as a brown gum (150 mg) according to the general procedure for urea formation (Method A), and used without further purification. LCMS: 439.5 [M+H]

[0342] [ka] Starting from 1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine (A1, 0.100 g, 0.332 mmol) and 1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-yl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)urea (0.146 g, 0.332 mmol), the title compound was obtained as an off-white solid (0.013 g, 8% yield) by following a procedure similar to that described for intermediate B26. 1 H NMR (400 MHz, DMSO-d6) δ = 11.68 (bs, 1H), 9.93 (bs, 1H), 8.53 (d, J = 2.0 Hz, 1H), 8.28 (s, 1H), 8.02-8.05 (m, 1H), 7.72 (d, J = 8.4 Hz, 1H), 6.27 (s, 1H), 3.86-3.90 (m, 1H), 1.10-1.49 (m, 8H).LCMS: 486.2 [M+H]

[0343] [ka] Starting with 3-(4-amino-3-methylphenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B27, 0.110 g, 0.392 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.135 g, 0.432 mmol), the title compound was obtained as a light brown solid (0.051 g, 26% yield) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, CD3OD) δ = 8.41 (s, 1H), 7.95-7.97 (m, 1H), 7.52-7.57 (m, 2H), 6.31 (s, 1H), 3.98-4.01 (m, 1H), 2.41 (s, 3H), 1.21-1.48 (m, 8H).LCMS: 499.1 [M+H]

[0344] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(1-methylazetidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B28, 0.130 g, 0.278 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.095 g, 0.306 mmol), the title compound was obtained as a white solid (0.010 g, yield 7%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, CD3OD) δ = 8.35-8.39 (m, 1H), 8.30 (s, 1H), 7.58-7.65 (m, 2H), 6.35 (s, 1H), 5.74-5.78 (m, 1H), 4.38-4.48 (m, 4H), 2.92 (s, 3H), 1.40-1.49 (m, 4H).LCMS: 532.2 [M+H]

[0345] [ka] Starting from 3-(4-amino-2,5-difluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B29, 0.050 g, 0.165 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.0516 g, 0.165 mmol), the title compound was obtained as an off-white solid (0.002 g, 3% yield) according to the general procedure for urea formation (Method B). 1H NMR (400 MHz, DMSO-d6) δ = 10.60 (bs, 1H), 9.08 (bs, 1H), 8.17 (s, 1H), 8.03-8.08 (m, 1H), 7.37-7.42 (m, 1H), 6.16 (s, 1H), 3.78-3.82 (m, 1H), 1.32-1.42 (m, 4H), 1.02-1.20 (m, 4H).LCMS: 521.3 [M+H]

[0346] [ka] Starting with (2-amino-5-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-3-yl)phenyl)methanol (B30, 0.120 g, 0.405 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.126 g, 0.405 mmol), the title compound was obtained as an off-white solid (0.025 g, yield 12%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, DMSO-d6) δ = 11.14 (bs, 1H), 8.65 (bs, 1H), 8.38 (s, 1H), 8.06-8.08 (m, 1H), 7.54-7.62 (m, 2H), 6.19 (s, 1H), 4.60 (s, 2H), 3.90-3.94 (m, 1H), 1.24-1.48 (m, 8H).LCMS: 515.2 [M+H]

[0347] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.120 g, 0.422 mmol) and phenyl (3-(2-cyanopropan-2-yl)isoxazole-5-yl)carbamate (C20, 0.126 g, 0.464 mmol), the title compound was obtained as an off-white solid (0.029 mg, yield 14%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, CD3OD) δ = 8.30-8.37 (m, 2H), 7.50-7.55 (m, 2H), 6.35 (s, 1H), 3.80-3.86 (m, 1H), 1.78 (s, 6H), 1.17-1.37 (m, 4H).LCMS: 462.2 [M+H]

[0348] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.100 g, 0.352 mmol) and phenyl (3-(((tert-butyldimethylsilyl)oxy)methyl)isoxazole-5-yl)carbamate (C21, 0.135 g, 0.387 mmol), the title compound was obtained as a white solid (0.020 g, yield 8%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, CD3OD) δ = 8.30-8.36 (m, 2H), 7.49-7.54 (m, 2H), 6.24 (s, 1H), 4.73 (s, 2H), 3.80-3.85 (m, 1H), 1.29-1.33 (m, 2H), 1.16-1.21 (m, 2H), 0.99 (s, 9H), 0.12 (s, 6H).LCMS: 539.3 [M+H]

[0349] [ka] TBAF (1 M, 0.037 ml, 0.037 mmol in THF) was added to a solution of 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-3-yl)-2-fluorophenyl)-3-(3-(((tert-butyldimethylsilyl)oxy)methyl)isoxazole-5-yl)urea (0.020 g, 0.037 mmol) in THF (3 ml) (0°C), and the resulting mixture was stirred at 25°C for 1 hour. After the reaction was complete (shown by TLC), the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (ELSD method, eluted with 1% TFA in a mixture of water and CAN) to obtain the title compound as a white solid (0.005 g, yield 31%, TFA salt). 1 H NMR (400 MHz, DMSO-d6) δ = δ 10.46 (bs, 1H), 8.98 (bs, 1H), 8.27-8.35 (m, 2H), 7.45-7.53 (m, 2H), 6.14 (s, 1H), 4.44 (s, 2H), 3.87-3.92 (m, 1H), 1.10-1.23 (m, 4H).LCMS: 425.2 [M+H]

[0350] [ka] Starting from 3-(6-aminopyridine-3-yl)-1-cyclopropyl-N-(2,4-dimethoxybenzyl)-1H-pyrazolo[4,3-c]pyridine-4-amine (B31, 0.050 g, 0.120 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.037 g, 0.120 mmol), the title compound was obtained as an off-white gum (0.012 g, yield 15%) according to the general procedure for urea formation (Method B). LCMS: 635.2 [M+H]

[0351] [ka] Triethylsilane (0.1 mL, 0.626 mmol) and TFA (0.1 mL, 1.298 mmol) were added to a solution of 1-(5-(1-cyclopropyl-4-((2,4-dimethoxybenzyl)amino)-1H-pyrazolo[4,3-c]pyridine-3-yl)pyridine-2-yl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)urea (0.012 g, 0.019 mmol) in DCM (2 mL) (0°C), and the resulting mixture was stirred at 25°C for 12 hours. After the reaction was complete (shown by LCMS), the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (eluted in ACN with 10 mM NH4OAc) to obtain the title product as an off-white solid (0.001 g, yield 10%). 1 H NMR (400 MHz, CD3OD) δ = 8.64 (d, J = 1.6 Hz, 1H), 8.09-8.11 (m, 1H), 7.80 (d, J = 6.0 Hz, 1H), 7.47-7.49 (m, 1H), 7.04 (d, J = 6.4 Hz, 1H), 6.39 (s, 1H), 3.80-3.86 (m, 1H), 0.90-1.50 (m, 8H).LCMS: 485.4 [M+H]

[0352] [ka] Starting with 2-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-d]pyrimidine-1-yl)ethane-1-ol (B32, 0.11 g, 0.382 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.10 g, 0.320 mmol), the title compound was obtained as an off-white solid (0.002 g, yield 1%) according to the general procedure for urea formation (Method B). 1H NMR (400 MHz, DMSO-d6) δ = 10.66 (bs, 1H), 8.99 (bs, 1H), 8.26-8.30 (m, 2H), 7.48-7.54 (m, 2H), 6.94 (bs, 2H), 6.22 (s, 1H), 4.90 (bs, 1H), 4.39 (t, J = 6.0 Hz, 2H), 3.85 (t, J = 6.0 Hz, 2H), 1.39-1.49 (m, 4H).LCMS: 507.2 [M+H]

[0353] [ka] Starting from 2-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-d]pyrimidine-1-yl)ethane-1-ol (B32, 0.100 g, 0.347 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.108 g, 0.347 mmol), the title compound was obtained as an off-white solid (0.004 g, yield 3%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, CD3OD) δ = 8.36-8.41 (m, 2H), 7.52-7.59 (m, 2H), 6.83 (s, 1H), 4.58 (t, J = 5.6 Hz, 2H), 4.07 (t, J = 5.2 Hz, 2H), 1.49-1.59 (m, 4H).LCMS: 507.2 [M+H]

[0354] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-(2-methoxyethyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B33, 0.125 g, 0.413 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.129 g, 0.413 mmol), the title compound was obtained as an off-white solid (0.020 g, 9% yield) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, DMSO-d6) δ = 10.61 (bs, 1H), 8.98 (bs, 1H), 8.25-8.29 (m, 2H), 7.48-7.54 (m, 2H), 6.96 (bs, 2H), 6.22 (s, 1H), 4.50 (t, LCMS: 521.2 [M+H]

[0355] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-(2-methoxyethyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B33, 0.125 g, 0.413 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.129 g, 0.413 mmol), the title compound was obtained as an off-white solid (0.015 g, 7% yield) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, DMSO-d6) δ = 10.07 (bs, 1H), 9.00 (bs, 1H), 8.31-8.35 (m, 2H), 7.47-7.55 (m, 2H), 6.92 (s, 1H), 4.53 (bs, 2H), 3.83 (t, J = 5.2 Hz, 2H), 3.23 (s, 3H), 1.51-1.52 (m, 4H).LCMS: 521.2 [M+H]

[0356] [ka] Starting with 3-(4-amino-3-methylphenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B27, 0.100 g, 0.357 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.123 g, 0.392 mmol), the title compound was obtained as a white solid (0.026 g, 15% yield) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, CD3OD) δ = 8.40 (s, 1H), 8.01-8.03 (m, 1H), 7.51-7.57 (m, 2H), 6.74 (s, 1H), 3.97-4.01 (m, 1H), 2.42 (s, 3H), 1.49-1.59 (m, 4H), 1.21-1.38 (m, 4H).LCMS: 499.5 [M+H]

[0357] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(3-(benzyloxy)cyclobutyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B26, 0.090 g, 0.223 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.076 g, 0.245 mmol), the title compound was obtained as a brown gum (70 mg) according to the general procedure for urea formation (Method B), and used without further purification. LCMS: 623.4 [M+H]

[0358] [ka] Boron trichloride (1 M, 0.899 mL, 0.899 mmol in DCM) was added dropwise to a solution of 1-(4-(4-amino-1-(3-(benzyloxy)cyclobutyl)-1H-pyrazolo[3,4-d]pyrimidine-3-yl)-2-fluorophenyl)-3-(5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)urea (0.070 g, 0.112 mmol) in 5 mL of dry DCM (-60°C), and the resulting mixture was incubated at 0°C for 3 hours. After the reaction was complete (shown by TLC and LCMS), the reaction mixture was cooled to -70°C, neutralized with aqueous ammonia (25% aqueous solution), and extracted with DCM (2 × 10 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by preparative HPLC (eluting with 1% TFA in a mixture of water and ACN) to obtain the title product as a white solid (0.009 g, yield 15%). 1 H NMR (400 MHz, CD3OD) δ = 8.36-8.40 (m, 2H), 7.53-7.61 (m, 2H), 6.82 (s, 1H), 5.62-5.69 (m, 1H), 4.74-4.80 (m, 1H), 2.94-2.99 (m, 2H), 2.58-2.63 (m, 2H), 1.49-1.58 (m, 4H).LCMS: 533.7 [M+H]

[0359] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-N-(2,4-dimethoxybenzyl)-1H-pyrazolo[4,3-c]pyridine-4-amine (B34, 0.220 g, 0.508 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.158 g, 0.508 mmol), the title compound was obtained as a brown gum (0.093 g, 28% yield) according to the general procedure for urea formation (Method B), and used without further purification. LCMS: 652.3 [M+H]

[0360] [ka] Triethylsilane (0.1 mL) and TFA (0.1 mL) were added to a solution of 1-(4-(1-cyclopropyl-4-((2,4-dimethoxybenzyl)amino)-1H-pyrazolo[4,3-c]pyridine-3-yl)-2-fluorophenyl)-3-(5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)urea (0.093 g, 0.143 mmol) in DCM (2 mL) (0°C), and the resulting mixture was stirred at 25°C for 12 hours. After the reaction was complete (shown by LCMS), the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (eluted with 0.1% TFA in a mixture of water and ACN) to obtain the title product as a white solid (0.050 g, yield 68%). 1 H NMR (400 MHz, DMSO-d6) δ = 10.10 (bs, 1H), 9.06 (bs, 1H), 8.33-8.37 (m, 1H), 7.89 (bs, 2H), 7.83 (d, J = 7.2 Hz, 1H), 7.54-7.57 (m, 1H), 7.44-7.47 (m, 1H), 7.32 (d, J = 7.2 Hz, 1H), 6.91 (s, 1H), 3.94-3.96 (m, 1H), 1.51-1.56 (m, 4H), 1.19-1.20 (m, 4H).LCMS: 502.5 [M+H]

[0361] [ka] Starting from 3-(4-amino-2,5-difluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B29, 0.100 g, 0.331 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.103 g, 0.331 mmol), the title compound was obtained as an off-white solid (0.002 g, yield 1%) according to the general procedure for urea formation (Method B). 1H NMR (400 MHz, CD3OD) δ = 8.24-8.29 (m, 2H), 7.38-7.43 (m, 1H), 6.85 (s, 1H), 3.80-3.85 (m, 1H), 1.47-1.59 (m, 4H), 1.20-1.33 (m, 4H).LCMS: 521.2 [M+H]

[0362] [ka] Starting with (2-amino-5-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-3-yl)phenyl)methanol (B30, 0.086 g, 0.290 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.091 g, 0.290 mmol), the title compound was obtained as an off-white solid (0.004 g, yield 3%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, DMSO-d6) δ = 10.39 (bs, 1H), 8.58 (bs, 1H), 8.27 (s, 1H), 8.06-8.08 (m, 1H), 7.52-7.62 (m, 2H), 6.89 (s, 1H), 5.45 (t, LCMS: 515.2 [M+H]

[0363] [ka] Starting from 3-(6-aminopyridine-3-yl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B35, 0.085 g, 0.318 mmol) and 1-phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.099 g, 0.318 mmol), the title compound was obtained as an off-white solid (0.003 g, yield 2%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, DMSO-d6) δ = 11.03 (bs, 1H), 9.83 (bs, 1H), 8.51 (bs, 1H), 8.27 (s, 1H), 8.01-8.04 (m, 1H), 7.72-7.74 (m, 1H), 6.97 (s, 1H), 3.86-3.88 (m, 1H), 1.49-1.58 (m, 4H), 1.12-1.24 (m, 4H).LCMS: 486.1 [M+H]

[0364] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B1, 0.100 g, 0.352 mmol) and phenyl (3-(tert-butyl)-4-methylisoxazole-5-yl)carbamate (C22, 0.096 g, 0.350 mmol), the title compound was obtained as a white solid (0.013 g, yield 8%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, CD3OD) δ = 8.39 (s, 1H), 8.22-8.26 (m, 1H), 7.46-7.55 (m, 2H), 3.94-3.97 (m, 1H), 2.09 (s, 3H), 1.35-1.39 (m, 11H), 1.21-1.23 (m, 2H).LCMS: 465.2 [M+H]

[0365] [ka] Starting with 1-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-d]pyrimidine-1-yl)-2-methylpropan-2-ol (B19, 0.100 g, 0.316 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.099 g, 0.316 mmol), the title compound was obtained as a white solid (0.033 g, yield 19%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, DMSO-d6) δ = 10.63 (bs, 1H), 8.97 (bs, 1H), 8.26-8.30 (m, 2H), 7.48-7.54 (m, 2H), 6.96 (bs, 2H), 6.22 (s, 1H), 4.77 (bs, 1H), 4.28 (s, 2H), 1.39-1.49 (m, 4H), 1.15 (s, 6H).LCMS: 535.3 [M+H]

[0366] [ka] Starting from 1-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-d]pyrimidine-1-yl)-2-methylpropan-2-ol (B19, 0.100 g, 0.316 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.099 g, 0.316 mmol), the title compound was obtained as a white solid (0.022 g, yield 13%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, DMSO-d6) δ = 10.08 (bs, 1H), 9.04 (bs, 1H), 8.42 (s, 1H), 8.31-8.35 (m, 1H), 7.47-7.55 (m, 2H), 6.91 (s, 1H), 4.31 (s, 2H), 1.48-1.58 (m, 4H), 1.17 (s, 6H).LCMS: 535.3 [M+H]

[0367] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(1-methylazetidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B28, 0.130 g, 0.278 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.095 g, 0.306 mmol), the title compound was obtained as a white solid (0.006 mg, yield 4%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, CD3OD δ = 8.26-8.41 (m, 2H), 7.56-7.64 (m, 2H), 6.84 (s, 1H), 5.71-5.74 (m, 1H), 4.29-4.90 (m, 4H), 2.85 (s, 3H), 1.52-1.59 (m, 4H).LCMS: 532.3 [M+H]

[0368] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B36, 0.06 g, 0.160 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.05 g, 0.160 mmol), the title compound was obtained as a light brown solid (80 mg) according to the general procedure for urea formation (Method A), and used without further purification. LCMS: 593.3 [M+H]

[0369] [ka] In dioxane, HCl (g) (4 M, 1 mL, 0.135 mmol) was added to a solution of 1-(4-(4-amino-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-d]pyrimidine-3-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)urea (0.08 g, 0.135 mmol) in DCM (5 mL) (0°C), and the resulting mixture was stirred at 25°C for 12 hours. After the reaction was complete (shown by UPLC), the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (eluted in ACN with 10 mM NH4OAc) to obtain the title product as an off-white solid (0.003 g, yield 5%). 1 H NMR (400 MHz, CD3OD) δ = 8.30-8.34 (m, 1H), 8.24 (s, 1H), 7.49-7.54 (m, 2H), 6.31 (s, 1H), 1.37-1.46 (m, 4H).LCMS: 463.3 [M+H]

[0370] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B36, 0.06 g, 0.160 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.05 g, 0.160 mmol), the title compound was obtained as a light brown solid (80 mg) according to the general procedure for urea formation (Method A), and used without further purification. LCMS: 593.3 [M+H]

[0371] [ka] In dioxane, HCl (g) (4 M, 1 mL, 0.135 mmol) was added to a solution of 1-(4-(4-amino-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-d]pyrimidine-3-yl)-2-fluorophenyl)-3-(5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)urea (0.08 g, 0.135 mmol) in DCM (5 mL) (0°C), and the resulting mixture was stirred at 25°C for 12 hours. After the reaction was complete (shown by ULC), the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (eluted with 10 mM NH4OAc in CAN) to obtain the title product as an off-white solid (0.007 g, yield 11%). 1 H NMR (400 MHz, DMSO-d6) δ = 13.61 (bs, 1H), 10.04 (bs, 1H), 8.96 (bs, 1H), 8.27-8.30 (m, 1H), 8.22 (s, 1H), 7.46-7.53 (m, 2H), 6.92 (s, 1H), 1.51-1.58 (m, 4H).LCMS: 463.3 [M+H]

[0372] [ka] Starting from 3-(4-amino-3-fluorophenyl)-1-(1-methylpiperidine-4-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B37, 0.100 g, 0.179 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazole-5-yl)carbamate (C6, 0.061 g, 0.197 mmol), the title compound was obtained as a white solid (0.008 g, yield 8%) according to the general procedure for urea formation (Method B). 1H NMR (400 MHz, CD3OD) δ = 8.34-8.38 (m, 2H), 7.52-7.57 (m, 2H), 6.35 (s, 1H), 5.12-5.17 (m, 1H), 3.73-3.76 (m, 2H), 3.33-3.39 (m, 2H), 3.00 (s, 3H), 2.59-2.62 (m, 2H), 2.35-2.39 (m, 2H), 1.45-1.49 (m, 4H).LCMS: 560.2 [M+H]

[0373] [ka] Starting with 3-(4-amino-3-fluorophenyl)-1-(1-methylpiperidine-4-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (B37, 0.100 g, 0.179 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazole-3-yl)carbamate (C7, 0.061 g, 0.197 mmol), the title compound was obtained as a brown solid (0.004 g, yield 4%) according to the general procedure for urea formation (Method B). 1 H NMR (400 MHz, CD3OD) δ = 8.36-8.40 (m, 2H), 7.51-7.56 (m, 2H), 6.82 (s, 1H), 5.10-5.18 (m, 1H), 3.73-3.76 (m, 2H), 3.37-3.39 (m, 2H), 3.00 (s, 3H), 2.59-2.62 (m, 2H), 2.35-2.38 (m, 2H), 1.39-1.59 (m, 4H).LCMS: 560.3 [M+H]

[0374] Biological Example 1 Compound biochemical assay Following the procedure described above, representative compounds were tested for their inhibitory activity against NEK7 and IL-1β release. The results are shown in the table below. [Table 29] [Table 30] Table 2 NEK7 IC 50 Regarding activity: * I C 50 is greater than 501 nM ** I C 50 The value is between 301 and 500 nM. *** I C 50 The value is between 151 and 300 nM. **** I C 50 is less than 150 nM Table 2 IL-1β IC 50 Regarding activity: + IC 50 is greater than 501 nM ++ IC 50 The value is between 201 and 500 nM. +++ IC 50 The value is between 51 and 200 nM. ++++ IC 50 is less than 50 nM - Indicates that the value could not be determined.

[0375] The various embodiments described above may be combined to provide further embodiments. All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent literature referenced herein and / or described in the application data sheets (including, but not limited to, U.S. Provisional Application No. 63 / 036,244 filed June 8, 2020, U.S. Provisional Application No. 63 / 170,776 filed April 5, 2021, and U.S. Provisional Application No. 63 / 185,257 filed May 6, 2021) are incorporated herein by reference as a whole. The aspects of the embodiments may be modified as necessary to provide further embodiments by utilizing concepts from various patents, applications and publications.

[0376] These and other modifications may be made to the embodiments in relation to the detailed description above. In general, the terms used in the following claims should not be construed as limiting the claims to the specific embodiments disclosed herein and herein, but rather as including all conceivable embodiments in addition to the entire scope of equivalents to which rights are granted. Thus, the claims are not limited by this disclosure.

Claims

1. Structure (I) below: 【Chemistry 1】 [In the formula, A is phenyl or pyridinyl, each containing one or more R ５ It may be replaced as appropriate; X is either N or CH; Y is NH; R １ is H or C １ -C ６ It is alkyl; R ２ is H, C １ -C ６ alkyl, C ２ -C ６ alkenyl, C ２ -C ６ alkynyl, C ３ -C ８ cycloalkyl, 3- to 8-membered heterocyclyl or 5- or 6-membered heteroaryl, each optionally substituted with one or more substituents selected from halo, hydroxyl, cyano, aminyl, C １ -C ６ alkyl, C ２ -C ６ alkenyl, C ２ -C ６ alkynyl, C １ -C ６ alkoxy and 3- to 8-membered heterocyclyl; R ３ The structure is as follows: 【Chemistry 2】 It is one of them; R ４ H is; R ５ These are, independently, Halo, Cyano, and C. １ -C ６ Alkyl, C １ -C ６ Hydroxylalkyl, C １ -C ６ Alkoxy or C １ -C ６ [It is a haloalkyl] A compound having, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

2. R ２ However, branch C ３ -C ６ Alkyl, C ２ -C ６ Alkenil, C ３ -C ６ Cycloalkyl, C ３ -C ８ A heterocyclyl or a 5- or 6-membered heteroaryl, each being a halo, hydroxyl, cyano, aminyl, or C. １ -C ６ Alkyl, C ２ -C ６ Alkenil, C ２ -C ６ Alkinyl, C １ -C ６ The compound according to claim 1, which may be optionally substituted with one or more substituents selected from alkoxys and 3 to 8-membered heterocyclines.

3. R ２ These are cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each being a halo, hydroxyl, cyano, aminyl, or C2 compound. １ -C ６ Alkyl, C ２ -C ６ Alkenil, C ２ -C ６ Alkinyl, C １ -C ６ The compound according to any one of claims 1 to 2, which may be optionally substituted with one or more substituents selected from alkoxys and 3 to 8-membered heterocyclines.

4. R ２ These are methyl, ethyl, isopropyl, 2-methylpropyl, or allyl, each being halo, hydroxyl, cyano, aminyl, or C. １ -C ６ Alkyl, C ２ -C ６ Alkenil, C ２ -C ６ Alkinyl, C １ -C ６ The compound according to any one of claims 1 to 2, which may be optionally substituted with one or more substituents selected from alkoxys and 3 to 8-membered heterocyclines.

5. R ２ These are oxetanil, tetrahydrofuranil, tetrahydropyranil, piperidinil, azetidinil, pyrrolidinil, or dioxidetetrahydrothiophenyl, each being a halo, hydroxyl, cyano, aminyl, or C compound. １ -C ６ Alkyl, C ２ -C ６ Alkenil, C ２ -C ６ Alkinyl, C １ -C ６ The compound according to any one of claims 1 to 2, which may be optionally substituted with one or more substituents selected from alkoxys and 3 to 8-membered heterocyclines.

6. R ２ However, it is pyridinyl, halo, hydroxyl, cyano, aminyl, C １ -C ６ Alkyl, C ２ -C ６ Alkenil, C ２ -C ６ Alkinyl, C １ -C ６ The compound according to any one of claims 1 to 2, which may be optionally substituted with one or more substituents selected from alkoxys and 3 to 8-membered heterocyclines.

7. R ２ However, the structure is as follows: 【Transformation 3】 The compound according to claim 1, having one of the following.

8. R ３ However, the structure is as follows: 【Chemistry 4】 A compound according to any one of claims 1 to 7, having one of the following.

9. R ４ The compound according to any one of claims 1 to 8, wherein H is present.

10. The compound according to claim 1, wherein A is phenyl.

11. The compound according to any one of claims 1 to 10, wherein A is unsubstituted.

12. A has one or more R ５ A compound according to any one of claims 1 to 10, which is substituted with

13. Each R ５ is fluoro, chloro, cyano, methyl, difluoromethyl, or -CH ２ OH, the compound according to claim 12.

14. A has the following structure: 【Transformation 5】 A compound according to any one of claims 1 to 13, having one of the following.

15. The following structure: 【Transformation 6】 【Transformation 7】 【Transformation 8】 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】 【Chemistry 15】 【Chemistry 16】 【Chemistry 17】 [Chemistry 18] A compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, having one of the above.

16. A pharmaceutical composition comprising a compound according to any one of claims 1 to 15, and a pharmaceutically acceptable carrier, diluent, or additive.

17. The pharmaceutical composition according to claim 16, which is useful as an NLRP3 inflammasome modulator, an NEK7 inhibitor, or both.

18. A pharmaceutical composition according to claim 17 for treating NLRP3-mediated disorders.

19. The pharmaceutical composition according to claim 18, wherein the disorder is selected from autoimmune disorders, inflammatory disorders, cardiovascular diseases, neurodegenerative disorders, bacterial and viral infections, allergies, asthma, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, transplantation, sperm motility, red blood cell deficiency, graft rejection, lung disorders, respiratory diseases, and ischemic conditions.

20. The pharmaceutical composition according to any one of claims 18 or 19, wherein the disorder is selected from type II diabetes, atherosclerosis, Alzheimer's disease, aging, fatty liver, metabolic syndrome, asthma, psoriasis, obesity, acute and chronic tissue injury due to infection, gout, arthritis, macular degeneration, enteritis, hepatitis, peritonitis, silicosis, sunburn due to UV rays, contact hypersensitivity, sepsis, cancer, neurodegenerative disease, multiple sclerosis, and Mackle-Wells syndrome.

21. A has the following structure: 【Chemistry 19】 The compound according to claim 1, having the following characteristics.

22. R ３ However, the structure is as follows: 【Chemistry 20】 A compound selected from the above, according to claim 1.

23. R ３ However, the structure is as follows: 【Chemistry 21】 A compound selected from the above, according to claim 1.

24. R ３ but, 【Chemistry 22】 The compound according to claim 1.

25. The compound has the following structure: 【Chemistry 23】 【Chemistry 24】 【Chemistry 25】 【Chemistry 26】 【Chemistry 27】 【Chemistry 28】 【Chemistry 29】 【Transformation 30】 The compound according to claim 1, having one of the following.

26. The compound has the following structure: 【Chemistry 31】 【Chemistry 32】 A compound according to claim 1, having one of the following, or a pharmaceutically acceptable salt or stereoisomer thereof.

27. The compound has the following structure: 【Transformation 33】 【Transformation 34】 A compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, having one of the above.

28. The compound has the following structure: 【Chemistry 35】 A compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, having one of the above.

29. The compound has the following structure: 【Transformation 36】 A compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, having one of the above.

30. The compound has the following structure: 【Chemistry 37】 A compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, having one of the above.

31. The compound has the following structure: 【Transformation 38】 The compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, having the above.

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