NEK7 inhibitors

Abstract

Compounds having activity as inhibitors of NEK7 are provided as compounds having the structure (I) or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein A, X, Y, Z, R 1 , R 2 , R 3 and n are defined herein. Methods for making and using these compounds, pharmaceutical compositions containing these compounds, and methods for modulating the activity of the NLRP3 inflammasome are provided.

Description

[Technical Field]

[0001] (Background technology) FIELD OF THE DISCLOSURE Embodiments of the present disclosure relate generally to compounds and methods of making and using them as therapeutic or prophylactic agents, for example, to treat inflammation.

[0002] 2. Description of Related Art Inflammasomes are multiprotein complexes whose activation plays a central role in innate immunity and inflammation. Four inflammasomes have been reported so far: NLRP1, NLRC4, NLRP3, and AIM2. The NLRP3 inflammasome is composed of NLRP3, ASC, and caspase-1. Its activation activates caspase-1, which promotes the secretion of IL-1β and IL-18, proinflammatory cytokines, in animal disease models of some autoimmune diseases, myocardial infarction, metabolic syndrome, inflammatory bowel disease, and macrophage activation syndrome.

[0003] NEK7 is a member of the NIMA-related kinase (NEK) family and acts as an NLRP3-binding protein, regulating NLRP3 oligomerization and activation. NEK7 is a serine / threonine kinase essential for mitotic entry, cell cycle progression, cell division, and mitotic progression, and is expressed in various tissues (e.g., brain, heart, lung, liver, and spleen). Overexpression of NEK7 leads to the production of abnormal cells, which are closely associated with tumors (e.g., retinoblastoma, gallbladder cancer, and head and neck cancer).

[0004] Numerous inhibitors have been widely used to inhibit effector signaling pathways involving IL-1β or IL-18 without abolishing inflammatory responses. NLRP3 inflammasome activation inhibitors that inhibit the NLRP3-NEK7 interaction may have therapeutic or preventive effects in some human diseases, such as type 2 diabetes (T2D), atherosclerosis, gout, and neurodegenerative diseases. However, the exact mechanism of NLRP3-NEK7 interaction remains unclear.

[0005] Thus, there is a need for the development of inhibitors that directly target NEK7 to affect inflammatory responses by modulating the NLRP3 inflammasome in certain pathological diseases (e.g., gout, atherosclerosis, type 2 diabetes, metabolic syndrome, macular degeneration, Alzheimer's disease, multiple sclerosis, and inflammatory bowel disease). Embodiments of the present disclosure fulfill this need and further provide related advantages. Summary of the Invention

[0006] Briefly, embodiments of the present disclosure provide compounds, pharmaceutically acceptable salts, stereoisomers, prodrugs, and the like, which are capable of modulating the activity of the NLRP3 inflammasome.

[0007] In one embodiment, the compound has structure (I): [ka] (In the formula, A, X, Y, Z, R 1 , R 2 , R 3 and n is each defined herein), or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof.

[0008] In another embodiment, pharmaceutical compositions comprising the disclosed compounds and methods of their use to treat inflammation are also provided. DETAILED DESCRIPTION OF THE INVENTION

[0009] In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments of the present disclosure. However, it will be understood by those skilled in the art that the present disclosure may be practiced without these details.

[0010] Unless the context otherwise requires, throughout this specification and claims, the word "comprise" and its variations, such as "comprises" and "comprising," are to be understood in their open and inclusive sense, including, but not limited to:

[0011] In this disclosure, any concentration range, percentage range, ratio range, or integer range is understood to include any integer in the recited range and, where appropriate, fractions thereof (e.g., tenths and hundredths of an integer), unless otherwise specified. As used herein, the terms "about" and "approximately" mean ±20%, ±10%, ±5%, or ±1% of the indicated range, value, or structure, unless otherwise specified. As used herein, the terms "a" and "an" should be understood to mean "one or more" of the listed components. When alternatives are used (e.g., "or"), it should be understood to mean either one of the alternatives, both, or any combination thereof.

[0012] Throughout this specification, the reference to "an embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. That is, the appearance of the phrase "in an embodiment" or "in an embodiment" in various places throughout this specification does not necessarily all refer to the same embodiment. Furthermore, the particular 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 commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the specification and claims, the use of the singular forms "a," "an," and "the" includes the plural unless the context clearly dictates otherwise.

[0014] "Amino" refers to the group -NH2. "Carboxy" or "carboxyl" refers to the group -CO2H. "Cyano" refers to the radical -CN. "Hydroxy" or "hydroxyl" refers to the group --OH. "Nitro" refers to the -NO2 group. "Oxo" refers to the =O substituent. "Thiol" refers to an --SH substituent. "Thioxo" refers to the =S substituent.

[0015] "Alkyl" refers to a saturated straight or branched chain hydrocarbon group consisting solely of carbon and hydrogen atoms, e.g., 1 to 12 carbon atoms (C1-C 12 "Ci-C alkyl," "Ci-C alkyl," "Ci-C alkyl," "Ci-C alkyl," or any value within these ranges (e.g., C4-C6 alkyl), and connected to the remainder 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, etc.). The carbon number referred to here refers to the main carbon chain and the carbon branch chains, and excludes carbon atoms contained in any substituents. Unless otherwise specified herein, alkyl groups may be optionally substituted.

[0016] "Alkenyl" means an unsaturated straight- or branched-chain hydrocarbon group, consisting solely of carbon and hydrogen atoms, containing one or more carbon-carbon double bonds, and having from 2 to 12 carbon atoms (C-C 12 Alkenyl refers to groups having 2 to 8 carbon atoms (C2-C8 alkenyl), 2 to 6 carbon atoms (C2-C6 alkenyl), or any number of carbon atoms within these ranges, attached to the remainder of the molecule by a single bond (e.g., ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1,4-dienyl, etc.). The carbon number referred to here refers to the main carbon chain and carbon branch chains and excludes carbon atoms contained in any substituents. Unless otherwise specified in this specification, alkenyl groups may be optionally substituted.

[0017] The term "alkynyl" refers to an unsaturated straight or branched chain hydrocarbon group containing 2 to 12 carbon atoms (C-C 12 The term "alkynyl" refers to a group having 2 to 9 carbon atoms (C2-C9 alkynyl), 2 to 6 carbon atoms (C2-C6 alkynyl), or any value within these ranges, and at least one carbon-carbon triple bond. Exemplary alkynyl groups may be selected from the group consisting of ethynyl, propargyl, but-1-ynyl, but-2-ynyl, and the like. The carbon number referred to here refers to the main carbon chain and the carbon branch chains, and excludes carbon atoms contained in any substituents. Unless otherwise specified in this specification, alkynyl groups may be optionally substituted.

[0018] "Alkoxy" means a group of the formula -OR a where R a is a group of 1 to 12 carbon atoms (C1-C 12 and alkyl groups as defined above containing 1 to 8 carbon atoms (C1-C8 alkoxy), 1 to 6 carbon atoms (C1-C6 alkoxy), or any value within these ranges. Unless stated otherwise in the specification, alkoxy groups may be optionally substituted.

[0019] "Aminyl" is a group of the formula -NR a R b where R a and R b are each independently H or C1-C6 alkyl as defined above. a and R b When both are H, the "aminyl" group is the same as the "amino" group defined above. Unless otherwise specified, the C1-C6 alkyl portion of the aminyl group may be optionally substituted.

[0020] "Aromatic ring" refers to a cyclic, planar molecule or portion of a molecule (i.e., group) having a conjugated ring system that exhibits enhanced stability compared to other bonding arrangements of atoms of the same type. Generally, aromatic rings contain a set of covalently bonded, coplanar atoms (e.g., alternating double and single bonds) and contain an even number of π-electrons that is not a multiple of four (i.e., 4n + 2 π-electrons, where n = 0, 1, 2, 3, etc.). Aromatic rings include, but are not limited to, phenyl, naphthenyl, imidazolyl, pyrrolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridonyl, pyridazinyl, and pyrimidonyl. Unless otherwise specified herein, "aromatic ring" includes all groups, optionally substituted.

[0021] "Aryl" refers to a carbocyclic group containing 6 to 18 carbon atoms, e.g., 6 to 10 carbon atoms (C6-C 10 The term "aryl" refers to a ring system having at least one carbon aromatic ring (aryl) and at least one carbon aromatic ring. In embodiments of the present disclosure, the aryl group is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, and may contain fused or bridged rings. Aryl groups include, but are not limited to, aryl groups derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Unless otherwise specified herein, the aryl group may be optionally substituted.

[0022] "Carbocyclic" or "carbocycle" refers to a ring system in which each ring atom is carbon.

[0023] "Cycloalkyl" refers to a non-aromatic monocyclic or polycyclic carbocyclic group, which may contain fused or bridged rings, consisting solely of carbon and hydrogen atoms, and which has 3 to 15 ring carbon atoms (C3-C 15 Cycloalkyl), 3 to 10 ring carbon atoms (C3-C 10"cycloalkyl" refers to a cycloalkyl group having 3 to 8 ring carbon atoms (C3-C8 cycloalkyl), or any value within these ranges (e.g., 3 to 4 carbon atoms (C3-C4 cycloalkyl)), saturated or partially unsaturated, and attached to the remainder of the molecule by a single bond. Monocyclic groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic groups include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise specified herein, cycloalkyl groups may be optionally substituted.

[0024] "Fused" refers to any ring structure described herein that is fused to another ring structure.

[0025] "Halo" refers to bromo, chloro, fluoro, or iodo.

[0026] "Haloalkyl" refers to an alkyl group, as defined above, substituted with one or more halogens, as defined above, such as trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc. Unless otherwise specified herein, a haloalkyl group may be optionally substituted.

[0027] "Halocycloalkyl" refers to a cycloalkyl group, as defined above, that is substituted with one or more halogens, as defined above, such as trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc. Unless otherwise specified in this specification, a halocycloalkyl group may be optionally substituted.

[0028] "Hydroxylalkyl" or "hydroxyalkyl" refers to an alkyl group, as defined above, that is substituted with one or more hydroxyl groups. The hydroxyalkyl group is attached to the backbone through an alkyl carbon atom. Unless otherwise specified herein, the hydroxyalkyl group may be optionally substituted.

[0029] "Heterocyclyl" refers to a 3- to 18-membered (e.g., 3- to 10-membered or 3- to 8-membered) non-aromatic ring group 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 specified herein, a heterocyclyl group is a partially or fully saturated monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused, spirocyclic, and / or bridged ring systems. The nitrogen, carbon, and sulfur atoms in a heterocyclyl group may be optionally oxidized, and the nitrogen atom may be optionally quaternized. Examples of such heterocyclyl groups include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, furanonyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, hexahydro-1H-pyrrolidine, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, oxiranyl, piperidinyl, piperazinyl, 4-piperidonyl, azetidinyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise in the specification, a heterocyclyl group may be optionally substituted.

[0030] "Heterocyclylalkyl" refers to a group containing a heterocyclyl group, as defined above, attached to the remainder of the molecule by an alkylene linker. Unless stated otherwise in the specification, the heterocyclylalkyl group may be optionally substituted.

[0031] A "heterocyclylcarbonyl" is a group of the formula -C(C=O)R a where R a is a heterocyclyl group as defined above. Unless stated otherwise in this specification, a heterocyclylcarbonyl group may be optionally substituted.

[0032] "Heterocyclyloxy" refers to a group of the formula -OR a where R a is a heterocyclyl group as defined above. Unless stated otherwise in this specification, a heterocyclyloxy group may be optionally substituted.

[0033] "Heterocyclylalkenyl" refers to a group of the formula -R a R b where R a is alkenylene, and R b is a heterocyclyl group as defined above. Unless stated otherwise in the specification, a heterocyclylalkenyl group may be optionally substituted.

[0034] "Heteroaryl" refers to a 5- to 18-membered (e.g., 5- or 6-membered) cyclic group containing 1 to 13 ring carbon atoms; 1 to 6 ring heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; and at least one aromatic ring. Heteroaryl groups can be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, and can include fused or bridged ring systems. In addition, the nitrogen, carbon, or sulfur atoms of the heteroaryl group can be optionally oxidized, and the nitrogen atom can be optionally quaternized. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, isothiazolyl, imidazolyl, indazolyl, indolyl, inda Examples of heteroaryl groups include zolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl). Unless stated otherwise in the specification, heteroaryl groups may be optionally substituted.

[0035] The suffix "-ene" refers to a specific structural feature (e.g., C6-C6) that is attached to the rest of the molecule through a single or double bond and to a group through a single or double bond. 10 Aryl, C3-C 10 The term "-ene" refers to a linker that has a structural feature in the moiety to which it is attached. The point of attachment of the "-ene" group to the rest of the molecule and to the group can be through one atom or any two atoms (carbon or heteroatom) within the group. For example, an unsubstituted phenylene group (i.e., typically C6-C6) 10 Arylene) has the following structure: [ka] It can be one of the following.

[0036] In some embodiments, C6-C 10 Arylene (e.g., phenylene), C3-C 10 The cycloalkylene (eg, cyclohexylene), 3- to 10-membered heterocyclylene (eg, piperidinylene), or 5- to 6-membered heteroarylene (eg, pyridinylene) is divalent.

[0037] As used herein, the term "substituted" refers to any of the above groups (e.g., alkyl, alkenyl, alkylene, alkylcarbonyl, alkoxy, alkoxyalkyl, aminylalkyl, aryl, cyanoalkyl, cycloalkyl, haloalkyl, heterocyclyl, heterocycle, heterocyclylalkyl, heteroaryl, heteroarylalkyl, and / or hydroxylalkyl) in which at least one hydrogen atom (e.g., one, two, three, or all hydrogen atoms) has been replaced with a non-hydrogen substituent. Examples of non-hydrogen substituents include, but are not limited to, amino, carboxyl, cyano, hydroxyl, halogen, nitro, oxo, thiol, thioxo, alkyl, alkenyl, alkylcarbonyl, alkoxy, aryl, cyanoalkyl, cycloalkyl, haloalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, and / or hydroxylalkyl, each optionally substituted with one or more of the above substituents.

[0038] In certain embodiments, the optional substituents are independently selected from halogen, hydroxyl, cyano, aminyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, C3-C8 halocycloalkyl, C6-C 10 It is selected from the group consisting of aryl, 5- or 6-membered heteroaryl, C1-C6 alkoxy, and 3- to 8-membered heterocyclyl.

[0039] The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound described herein sufficient to achieve the intended effect (including, but not limited to, the treatment of diseases defined below). A therapeutically effective amount may vary depending on the intended therapeutic use (in vivo), the subject and condition being treated (e.g., the subject's weight and age, the severity of the condition), the method of administration, etc., and can be readily determined by one skilled in the art. The term also applies to a dose that produces a specific response in target cells (e.g., decreased platelet adhesion and / or decreased cell migration). The specific dose will vary depending on the specific compound selected, the subsequent administration regimen, whether it is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the delivery system in the body by which the compound is delivered.

[0040] As used herein, "treatment" or "treating" refers to a method for achieving a beneficial or desired result (including, but not limited to, a therapeutic effect and / or a prophylactic effect) with respect to a disease, disorder, or condition. Therapeutic efficacy refers to the eradication or amelioration of the underlying disease being treated. Therapeutic efficacy can also be achieved by eradicating or ameliorating one or more physiological symptoms associated with the underlying disease, to the extent that the subject remains afflicted with the underlying disease, to the extent that an improvement is observed in the subject. A prophylactic effect can include delaying or preventing the onset of a disease or condition, delaying or preventing the onset of a disease or condition, delaying, halting, or reversing the progression of a disease or condition, or any combination thereof. In certain embodiments, a composition is administered prophylactically to a subject at risk of developing a particular disease, or to a subject who has reported one or more physiological symptoms of a disease, even if the disease has not been diagnosed.

[0041] As used herein, the terms "co-administration," "administration in combination," and similar expressions include administering two or more agents to an animal, such as a human, such that both agents and / or their metabolites are present in the subject at the same time. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in a composition containing both agents.

[0042] "Pharmaceutically acceptable salt" includes both acid and base addition salts.

[0043] A "pharmaceutically acceptable acid addition salt" refers to a salt having a biologically effective free base, which is a biologically acceptable salt or is biologically suitable for administration to a subject. See generally 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. Preferred pharmaceutically acceptable acid addition salts are pharmacologically effective and suitable for contact with patient tissues without undue toxicity, irritation, or allergic response. Pharmaceutically acceptable acid addition salts include those with inorganic acids (such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.), and organic acids (such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, 10-camphorsulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfonic acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronolic acid, methylparaben ... acid, glutamic acid, glutaric acid, 2-oxoglutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic 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, etc.

[0044] A "pharmaceutically acceptable base addition salt" refers to a salt with a biologically effective free acid that is biologically acceptable or biologically suitable for administration to a subject. See, for example, 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. Preferred pharmaceutically acceptable base addition salts are pharmacologically effective and suitable for contact with patient tissues without undue toxicity, irritation, or allergic reaction. Pharmaceutically acceptable base addition salts are prepared by adding an inorganic or organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, naturally occurring substituted amines, cyclic amines, and substituted amines such as 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, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc.). Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0045] In some embodiments, pharmaceutically acceptable salts include quaternary ammonium salts (salts of quaternary amine alkyl halides such as methyl bromide).

[0046] The terms "antagonist" and "inhibitor" are used interchangeably and refer to compounds that can inhibit the biological function of a target protein by inhibiting the activity or expression of the protein (e.g., NLRP3 inflammasome or NEK7) or the association of NLRP3 inflammasome with NEK7. Thus, the terms "antagonist" and "inhibitor" are defined within the biological role of the target protein. Preferred antagonists herein specifically interact (e.g., bind) with the target, but compounds that inhibit the biological activity of the target protein by interacting with other members of the signal transduction pathway of which the target protein is a member are specifically included within this definition. Preferred biological activities inhibited by antagonists are those related to tumor development, growth, and spread.

[0047] As used herein, the term "agonist" refers to a compound that can activate or enhance the biological function of a target protein by inhibiting the activity or expression of the target protein. Thus, the term "agonist" is defined within the context of the biological role of the target polypeptide. Preferred agonists herein specifically interact (e.g., bind) with the target, although compounds that activate or enhance the biological activity of a target polypeptide by interacting with other members of a signal transduction pathway of which the target polypeptide is a member are specifically included in this definition.

[0048] "Signal transduction" refers to the series of processes by which stimulatory or inhibitory signals are transmitted to a cell to produce a response within the cell.

[0049] The terms "selective inhibition" or "selectively inhibit" refer to the ability of a bioactive agent, through direct or indirect interaction with a target, to selectively reduce the activity of transducing a target signal relative to the activity of transducing an off-target signal.

[0050] "Subject" refers to an animal (e.g., a mammal, such as a human). The methods described herein can be useful for both human treatment and veterinary applications. In some embodiments, the subject is a mammal, and in some embodiments, the subject is a human.

[0051] "Mammals" includes humans and domestic animals (eg, laboratory animals and pets such as cats, dogs, pigs, cows, sheep, goats, horses, rabbits, etc.), and non-domestic animals (eg, wild animals, etc.).

[0052] "Prodrug" means a compound that can be converted under physiological conditions or by solvolysis into a biologically active compound described herein (e.g., a compound of structure (I)). Thus, the term "prodrug" refers to a pharmaceutically acceptable precursor of a biologically active compound. In some embodiments, a prodrug is inactive when administered to a subject, but is converted to an active compound in vivo (e.g., by hydrolysis). Such prodrug compounds often offer advantages in solubility, mammalian tissue compatibility, or delayed release (see, e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam)). For a discussion of prodrugs, see Higuchi, T., et al., "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 incorporated herein by reference in their entirety. The term "prodrug" is also meant to include any covalently bonded carrier that releases the active compound in vivo when the prodrug is administered to a mammalian subject. Prodrugs of the active compounds described herein are generally prepared by modifying functional groups in the active compound such that the modification cleaves, either by routine manipulation or in vivo, to yield the original active compound. Prodrugs include compounds in which a hydroxy, amino, or thiol group in the compound is bonded to any group that cleaves to yield a free hydroxy, amino, or mercapto group, respectively, when the prodrug of the active compound is administered to a mammalian subject. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of hydroxy groups, or acetamide, formamide, and benzamide derivatives of amino groups in the active compound, etc.

[0053] The term "in vivo" refers to an event that takes place inside a subject's body.

[0054] Embodiments of the present disclosure are also meant to include all pharmaceutically acceptable compounds of structure (I), salts, stereoisomers, tautomers, polymorphs, solvates, hydrates, and prodrugs thereof.

[0055] Certain embodiments are also meant to include the in vivo metabolic products of the disclosed compounds. Such products may result, primarily through enzymatic action, from, for example, oxidation, reduction, hydrolysis, amidation, esterification, etc., of the administered compound. Accordingly, embodiments include compounds obtained by a method comprising administering a compound of the present disclosure to a mammal for a period of time sufficient to yield the metabolic products. Such products are generally identified by administering to an animal (e.g., a rat, mouse, guinea pig, monkey, or human) a detectable dose of an isotopically labeled compound of the present disclosure, allowing sufficient time for metabolism to occur, and isolating the converted product from urine, blood, or other biological sample.

[0056] By "stable compound" and "stable structure" is meant a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

[0057] Crystallization often results in solvates of the compounds of the present disclosure. As used herein, the term "solvate" refers to an aggregate comprising one or more compounds of the present disclosure and one or more solvent molecules. In some embodiments, the solvent is water, in which case the solvate is a hydrate. Alternatively, in other embodiments, the solvent is an organic solvent. Therefore, the compounds of the present disclosure exist as hydrates, such as monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, or corresponding solvates. In some embodiments, the compounds of the present disclosure are actually solvates, while in other embodiments, the compounds of the present disclosure merely contain incidental water or are a mixture of water and some incidental solvent.

[0058] "Optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and includes cases where the event or circumstance occurs or not. For example, "optionally substituted aryl" means that the aryl group may or may not be substituted, and includes both substituted and unsubstituted aryl groups.

[0059] A "pharmaceutical composition" refers to a formulation containing a compound of the present disclosure and a vehicle generally accepted in the art for delivering the compound of the present disclosure to a mammal (e.g., a human). Such vehicles include any pharmaceutically acceptable carrier, diluent, or excipient therefor.

[0060] A "pharmaceutically acceptable carrier, diluent, or excipient" includes, but is not limited to, any adjuvant, carrier, excipient, flow agent, sweetener, diluent, preservative, colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier.

[0061] "Stereoisomers" refer to compounds composed of the same atoms and bonds but with different, non-interchangeable three-dimensional structures. The present disclosure contemplates various stereoisomers and mixtures thereof, including "enantiomers." Enantiomers refer to two stereoisomers whose molecules are non-superimposable mirror images of one another.

[0062] The compounds of the present disclosure (i.e., compounds of structure (I)) or pharmaceutically acceptable salts thereof may contain one or more centers of geometric asymmetry and thus may give rise to stereoisomers (e.g., enantiomers, diastereomers, and other stereoisomers) defined in absolute stereochemistry as (R)- or (S)-, or for amino acids, (D)- or (L)-. Thus, embodiments include all possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthetic equivalents or chiral reagents or resolved by conventional techniques (e.g., chromatography and fraction crystallization). Conventional techniques for the preparation / separation of individual enantiomers include chiral synthesis from appropriate optically pure precursors or resolution of the racemate (or racemate of a salt or derivative) using, for example, chiral high-performance liquid chromatography (HPLC). If the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, the compounds are intended to include both E and Z geometric isomers, as well as all tautomeric forms.

[0063] Embodiments of the present disclosure include all rotamers and conformationally restricted states of the disclosed compounds. Also included are atropisomers, which are stereoisomers resulting from hindered rotation around a single bond, where the energy difference due to steric strain or other contributing factors creates a rotational barrier large enough to separate the individual conformers. By way of example, certain compounds of the present disclosure may exist as a mixture of atropisomers, or as a single pure atropisomer, or as a state enriched in one atropisomer.

[0064] 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 one enantiomer or diastereomer.

[0065] "Tautomer" refers to the migration of a proton from one atom of a molecule to another atom of the same molecule. Thus, embodiments include tautomers of the disclosed compounds.

[0066] The chemical nomenclature and structural formulae used herein have been adapted from 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). Complex chemical names used herein are generally named so that the name of the substituent precedes the group to which it is attached. For example, cyclopropylethyl contains an ethyl backbone and a cyclopropyl substituent. Except as noted below, in the chemical structures used herein, all bonds, except for those on select carbon atoms, are identified and assumed to be connected to sufficient hydrogen atoms to satisfy valence.

[0067] (compound) The present disclosure provides compounds, pharmaceutically acceptable salts, stereoisomers, prodrugs, and the like, which are capable of modulating the activity of the NLRP3 inflammasome.

[0068] An embodiment of the present disclosure is a compound having the following structure (I): [ka] [In the formula, A is C6-C 10 Arylene, C3-C 10 cycloalkylene, 3- to 10-membered heterocyclylene, or 5- to 6-membered heteroarylene; X is N or CR 4 and; Y is N or CH; R 1 is C1-C6 alkyl, C1-C6 hydroxyl alkyl, C3-C 10 cycloalkyl, or 3-10 membered heterocyclyl; R 2is a 3- to 10-membered heterocyclyl, a 3- to 10-membered heterocyclylalkyl, a 3- to 10-membered heterocyclylalkenyl, a 3- to 10-membered heterocyclylcarbonyl, a 3- to 10-membered heterocyclyloxy, or a 5- to 6-membered heteroaryl; R 3 each, if present, is independently halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or C3-C8 cycloalkyl; R 4 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C3-C8 cycloalkyl; and n is 0, 1, 2, 3, or 4] or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof.

[0069] Another embodiment is a compound having the following structure (I): [ka] [In the formula, A is C6-C 10 Arylene, C3-C 10 cycloalkylene, 3- to 10-membered heterocyclylene, or 5- to 6-membered heteroarylene; X is N or CR 4 and; Y is N or CH; Z is N or CH; R 1 is C1-C6 alkyl, C1-C6 alkynyl, C1-C6 hydroxylalkyl, C1-C6 carboxyalkyl, C1-C6 alkoxyalkyl, C3-C 10 cycloalkyl, or 3-10 membered heterocyclyl; R 2 is a 3- to 10-membered heterocyclyl, a 3- to 10-membered heterocyclylalkyl, a 3- to 10-membered heterocyclylalkenyl, a 3- to 10-membered heterocyclylcarbonyl, a 3- to 10-membered heterocyclyloxy, or a 5- to 6-membered heteroaryl; R 2 is R 3 If there is R 2 R bonded to the carbon adjacent to the carbon to which 3 and combine to form a C3-C8 cycloalkyl; R 3 each, if present, is independently halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or C3-C8 cycloalkyl; R 4 is H, C1-C6 alkyl, C1-C6 haloalkyl, or C3-C8 cycloalkyl; and n is 0, 1, 2, 3, or 4] or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof.

[0070] In some embodiments, A is C6-C 10 In certain embodiments, A is arylene. In certain embodiments, A is phenylene. In certain embodiments, A is 5-6 membered heteroarylene. In certain embodiments, A is pyridinylene. In certain embodiments, A is C3-C 10 In certain embodiments, A is substituted with one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or C3-C8 cycloalkyl. In certain embodiments, A is substituted with one or more halogen substituents. In other embodiments, A is unsubstituted.

[0071] In some embodiments, X is CR 4 In yet another embodiment, R 4 is H or C1-C6 alkyl. In some embodiments, R 4 is H. In certain embodiments, X is N.

[0072] In some embodiments, R 1 is C1-C6 alkyl. In yet another particular embodiment, R 1 is methyl or isopropyl. In certain embodiments, R 1 is C1-C6 hydroxylalkyl. In a more particular embodiment, R 1 has the following structure: [ka] It has one of the following.

[0073] In some embodiments, R 1 has the following structure: [ka] It has one of the following.

[0074] In some embodiments, R 1 is C1-C6 carboxyalkyl. In still other embodiments, R 1 has the following structure: [ka] It has one of the following.

[0075] In some embodiments, R 1 is C1-C6 alkoxyalkyl. In some embodiments, R 1 has the following structure: [ka] It has.

[0076] In some embodiments, R 1 is C3-C 10 In yet another embodiment, R 1 is cyclopropyl or cyclobutyl. In certain embodiments, R 1 is 3-10 membered heterocyclyl. In certain embodiments, R 1is oxetanyl, pyrrolidinyl, or piperidinyl. 1 is oxetanyl, pyrrolidinyl, azetidinyl, or piperidinyl.

[0077] In some embodiments, R 1 is C1-C6 alkynyl. In certain embodiments, R 1 has the following structure: [ka] It has one of the following.

[0078] In some embodiments, R 1 is substituted with one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or C3-C8 cycloalkyl. 1 is substituted with one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, -S(O)2CH3, -S(O)2 cyclopropyl, or C3-C8 cycloalkyl. 1 is substituted with one or more C1-C6 alkyl substituents. 1 is not substituted.

[0079] In certain embodiments, R 1 has the following structure: [ka] It has one of the following.

[0080] In some embodiments, R 1 has the following structure: [ka] It has one of the following.

[0081] In certain embodiments, the compound or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof has the following structure (Ia): [ka] It has.

[0082] In certain embodiments, the compound or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof has the following structure (Ia): [ka] It has.

[0083] In some embodiments, Y is N. In other embodiments, Y is CH.

[0084] In certain embodiments, the compound or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof has the following structure (Ib): [ka] It has.

[0085] In certain embodiments, the compound or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof has the following structure (Ib): [ka] It has.

[0086] In some embodiments, R 2 is a 3- to 10-membered heterocyclyl. 2 is morpholino. In other particular embodiments, R 2 is piperazinyl. In yet another particular embodiment, R 2 has the following structure: [ka] It has one of the following.

[0087] In some embodiments, R 2 is 3-10 membered heterocyclylalkyl. In more particular embodiments, R 2 has the following structure: [ka] It has one of the following.

[0088] In yet other particular embodiments, R 2 has the following structure: [ka] It has one of the following.

[0089] In some embodiments, R 2 is a 3- to 10-membered heterocyclylcarbonyl. 2 has the following structure: [ka] It has.

[0090] In some embodiments, R 2 is a 5-6 membered heteroaryl. In yet other particular embodiments, R 2 has the following structure: [ka] It has.

[0091] In some embodiments, R 2 is 3-10 membered heterocyclyloxy. 2 has the following structure: [ka] It has.

[0092] In some embodiments, R 2 has the following structure: [ka] It has one of the following.

[0093] In yet other particular embodiments, R 2 has the following structure: [ka] It has.

[0094] In some embodiments, n is 0. In some embodiments, n is 1 or 2. In some embodiments, n is 1. In some embodiments, R 3 is halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or C3-C8 cycloalkyl. 3 is methyl, chloro, fluoro, cyano, difluoromethyl, trifluoromethyl, methoxy, trifluoromethoxy, or cyclopropyl. In certain embodiments, n is 1 or 2 and R 3 is halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or C3-C8 cycloalkyl. In some embodiments, n is 1 and R 3 is halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or C3-C8 cycloalkyl. In some embodiments, n is 1 and R 3 is methyl, chloro, fluoro, trifluoromethyl, methoxy, trifluoromethoxy, or cyclopropyl.

[0095] In some embodiments, the compound or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof has the following structure (Ic): [ka] [In the formula, R 3a is halogen or 3- to 10-membered heterocyclyl; and n1 is 1, 2, or 3.

[0096] In some embodiments, R 3a is fluoro or piperazinyl. In still other embodiments, the compound or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof has the following structure (Ic1): [ka] It has.

[0097] In certain embodiments, the compound of structure (I) is a modulator of the NLRP3 inflammasome. In some embodiments, the compound of structure (I) is a modulator of the NLRP3 inflammasome in a patient or biological sample.

[0098] In certain embodiments, the compound of structure (I) is an inhibitor of NEK7. In some embodiments, the compound of structure (I) is an inhibitor of NEK7 in a patient or biological sample.

[0099] In various embodiments, the compound, or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof, has one of the structures shown below in Table 1. The compounds in Table 1 may be prepared according to the Examples or methods known to those skilled in the art and analyzed by mass spectrometry and / or 1 Analyzed by 1 H 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 17

Table 18

Table 19

[0100] It is understood that combinations of substituents and / or variables of the depicted formulae are permissible herein only if such combinations result in stable compounds.

[0101] In further embodiments, various compounds of the present disclosure that exist in free base or free acid form can be converted to a pharmaceutically acceptable salt by treatment with an appropriate inorganic or organic base or acid by methods known to those skilled in the art. Salts of compounds of the present disclosure can be converted to the free base or free acid form by standard techniques.

[0102] Pharmaceutical Composition Other embodiments relate to pharmaceutical compositions. The pharmaceutical composition comprises any 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 a compound of the present disclosure and another therapeutic agent (e.g., an anti-cancer agent). By way of example and not limitation, such another therapeutic agent is described below.

[0103] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ocular, pulmonary, transmucosal, transdermal, vaginal, otic, nasal, and topical administration. Further, by way of example only, parenteral administration includes intramuscular, subcutaneous, intravenous, intrathecal injection, as well as intrathecal, direct intraventricular, intraperitoneal, intralymphatic, and intranasal injection.

[0104] In some embodiments, the compounds described herein are administered locally rather than systemically (e.g., by injecting the compound directly into an organ, often in the form of a depot or sustained-release formulation). In certain embodiments, long-acting formulations are administered by implantation (e.g., subcutaneously or intramuscularly) or intramuscular injection. Furthermore, in other embodiments, the compounds are delivered in targeted drug delivery systems (e.g., liposomes coated with organ-specific antibodies). In the above embodiments, the liposomes are selectively taken up by the organ. In yet another embodiment, the compounds described herein are provided in the form of immediate-release, sustained-release, or intermediate-release formulations. In yet another embodiment, the compounds described herein are administered locally.

[0105] In the therapeutic methods of embodiments of the present disclosure, a subject suffering from or diagnosed with a disease, disorder, or condition is administered an effective amount of at least one compound of structure (I). Effective amounts or doses may be ascertained by methods such as modeling, dose escalation studies, or clinical trials (e.g., taking into account the method or route of administration or drug delivery, the pharmacokinetics of the agent, the severity and course of the disease, disorder, or condition, previous or current treatments of the subject, the subject's health status and response to drugs, and the judgment of the treating physician).

[0106] The compounds described herein are effective over a wide range of dosages. For example, in some embodiments, dosages of 10-5000 mg, 100-5000 mg, or 1000 mg-4000 mg per day are used, with 1000-3000 mg per day being examples of dosages used in treating adult humans. The exact dosage will vary depending on the route of administration, the form of the compound administered, the subject being treated, the subject's weight, and the wishes and experience of the treating physician.

[0107] In some embodiments, the compound of the present disclosure is administered in a single dose.The single dose is generally administered by injection (for example, intravenous injection) to allow the drug to be rapidly administered, although other routes may also be used as appropriate.The single dose of the compound of the present disclosure can also be used to treat acute symptoms.

[0108] In some embodiments, the compound of the present disclosure is administered multiple times. In some embodiments, administration is about once, twice, three times, four times, five times, six times, or more than six times per day. In other embodiments, administration is about once a month, once every two weeks, once a week, or once every other day. In another embodiment, the compound of the present disclosure and another agent (e.g., an anticancer agent) are administered together about once to about six times per day. In another embodiment, administration of the compound of the present disclosure and another agent continues for less than about 7 days. In yet another embodiment, the administration continues for more than about 6 days, 10 days, 14 days, 28 days, 2 months, 6 months, or 1 year. In some cases, continuous administration is continued as long as necessary.

[0109] The administration of the compound of the present disclosure can be continued as long as necessary.In some embodiments, the compound of the present disclosure is administered for a period of 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 14 days or more than 28 days.In some embodiments, the compound of the present disclosure is administered for less than 28 days, 14 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days or less than 1 day.In some embodiments, the compound of the present disclosure is administered continuously for a long period of time, for example, to treat ongoing chronic symptoms.

[0110] In some embodiments, the compounds of the present disclosure are administered in individual dosage forms. It is known in the art that subject-to-subject variability in the pharmacokinetics of compounds necessitates individualization of dosing regimens for optimal treatment.

[0111] In some embodiments, the compounds described herein are formulated into pharmaceutical compositions.In certain embodiments, pharmaceutical compositions are formulated by conventional methods using one or more physiologically acceptable carriers, including excipients and auxiliaries that facilitate the disclosed compounds to be formulated for pharmaceutical use.Suitable formulations vary according to the route of administration selected.Any pharmaceutically acceptable method, carrier, and excipient can be used to formulate pharmaceutical compositions as described herein, Remington, The Science and Practice of Pharmacy, Nineteenth Edition (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 Edition (Lippincott Williams & Wilkins 1999).

[0112] Provided herein are pharmaceutical compositions comprising one or more compounds of structure (I) and a pharmaceutically acceptable carrier.

[0113] Provided herein is a pharmaceutical composition comprising one or more compounds selected from the compounds of structure (I) and pharmaceutically acceptable diluents, excipients, and carriers.In some embodiments, the compounds described are administered as a combination therapy pharmaceutical composition, in which one or more compounds selected from the compounds of structure (I) are mixed with other active ingredients.The present disclosure encompasses any combination of active ingredients as set forth in the combination therapy section below and throughout this disclosure.In certain embodiments, the pharmaceutical composition comprises one or more compounds of structure (I).

[0114] In certain embodiments, the pharmaceutical composition of the compound of structure (I) is a modulator of the NLRP3 inflammasome.

[0115] In certain embodiments, pharmaceutical compositions of compounds of structure (I) inhibit NEK7 when administered to a patient or biological sample.

[0116] As used herein, a pharmaceutical composition refers to a mixture of one or more compounds selected from the compounds of structure (I) with other chemical components (e.g., carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients). In certain embodiments, the pharmaceutical composition facilitates administration of the compound to an organism. In some embodiments, a therapeutically effective amount of one or more compounds selected from the compounds of structure (I) provided herein is administered as a pharmaceutical composition to a mammal with a disease, disorder, or condition to be treated. In certain embodiments, the mammal is a human. In certain embodiments, the therapeutically effective amount will vary 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.

[0117] In certain embodiments, one or more compounds selected from the compounds of structure (I) are formulated in an aqueous solution. In certain embodiments, the aqueous solution is selected from a physiologically compatible buffer, such as, by way of example only, Hank's solution, Ringer's solution, or physiological saline. In other embodiments, one or more compounds selected from the compounds of structure (I) are formulated for transmucosal administration. In certain embodiments, the transmucosal formulation includes a penetrant appropriate for the barrier that must be permeated. In embodiments in which the compounds described herein are formulated for other parenteral injections, suitable formulations include aqueous or non-aqueous solutions. In such embodiments, the solution includes a physiologically compatible buffer and / or excipient.

[0118] In another embodiment, the compound described herein is formulated for oral administration.The compound described herein is formulated by combining active compound with, for example, pharmaceutically acceptable carrier or excipient.In various embodiments, the compound described herein is formulated as oral preparation (for example, tablet, powder, pill, dragee, capsule, liquid, gel, syrup, elixir, slurry, suspension etc.).

[0119] In some embodiments, oral pharmaceutical preparations can be obtained by mixing one or more solid excipients with one or more compounds described herein, optionally grinding the resulting mixture, and then processing the resulting granule mixture after adding suitable excipients, to obtain the core of tablets or dragees.Suitable excipients include, in particular, fillers (such as sugars such as lactose, sucrose, mannitol, or sorbitol); celluloses (such as cereal starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose); or others (such as polyvinylpyrrolidone (PVP or povidone) or calcium phosphate).In certain embodiments, disintegrants can be optionally added.Disintegrants include, by way of example only, cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or its salt (such as sodium alginate).

[0120] In some embodiments, dosage forms (e.g., dragee cores and tablets) are provided with one or more suitable coatings. In certain embodiments, concentrated sugar solutions are used to coat dosage forms. The sugar solutions include, for example, gum arabic, talc, polyvinylpyrrolidone, carbopol, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures, among other additives. Dyes and / or pigments are also optionally added to the coating for identification. Furthermore, the dyes and / or pigments are optionally used to distinguish different combinations of active compound doses.

[0121] In some embodiments, a therapeutically effective amount of at least one compound described herein is formulated into other oral dosage forms. Oral dosage forms include filled capsules made of gelatin and soft, sealed capsules made of gelatin and a plasticizer (e.g., glycerol or sorbitol). In certain embodiments, the filled capsules contain a mixture of active ingredients and one or more fillers. Fillers include, by way of example only, lactose, binders (e.g., starch), and / or lubricants (e.g., talc or magnesium stearate), and optionally, stabilizers. In other embodiments, the soft capsules contain one or more active compounds dissolved or suspended in a suitable liquid. Suitable liquids include, by way of example only, one or more fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers are optionally added.

[0122] In yet other embodiments, the compounds described herein are formulated for parenteral administration, including suitable formulations such as bolus injection or continuous infusion. In certain embodiments, the injectable formulations are in single-dose form (e.g., ampoules) or in multi-dose containers. Preservatives are optionally added to the injectable formulations. In still other embodiments, the pharmaceutical composition is formulated as a sterile suspension, solution, or emulsion in an oily or aqueous vehicle in a form suitable for parenteral injection. Parenteral injection preparations may optionally contain formulating agents such as suspending agents, stabilizers, and / or dispersing agents. In certain embodiments, the pharmaceutical formulation for parenteral administration comprises an aqueous solution of a water-soluble active compound. In another embodiment, a suspension of one or more compounds selected from the compounds of structure (I) is prepared as a suitable oily injection suspension. Suitable lipophilic solvents or vehicles for use in the pharmaceutical compositions described herein include, by way of example only, fatty oils (e.g., sesame oil), or synthetic fatty acid esters (e.g., ethyl oleate or triglycerides), or liposomes. In certain embodiments, aqueous injection suspensions contain substances that increase the viscosity of the suspension (for example, sodium carboxymethylcellulose, sorbitol, or dextran).The suspension may optionally contain suitable stabilizers or substances that increase the solubility of the compound in order to prepare highly concentrated solutions.Alternatively, in other embodiments, the active ingredient is in powder form before use, so that it is prepared in a suitable vehicle (for example, sterile pyrogen-free water).

[0123] Pharmaceutical compositions include at least one pharmaceutically acceptable carrier, diluent, or excipient and, as an active ingredient, one or more compounds selected from the compounds of structure (I) described herein. The active ingredient may be in the form of a free acid or free base, or a pharmaceutically acceptable salt. Additionally, the methods and pharmaceutical compositions described herein include the use of N-oxides, crystalline forms (also known as polymorphs), and active metabolites of these compounds, which have similar activity. All tautomers of the compounds described herein are included within the scope of the compounds described herein. Furthermore, the compounds described herein include unsolvated forms as well as solvated forms formed with pharmaceutically acceptable solvents (e.g., water, ethanol, etc.). Solvated forms of the compounds described herein are also considered to be disclosed herein. Additionally, the pharmaceutical compositions may optionally include other medicinal or pharmaceutical substances, carriers, adjuvants (e.g., preservatives, stabilizers, wetting or emulsifying agents, solubility enhancers, salts for adjusting osmotic pressure, buffers, and / or other therapeutically effective substances).

[0124] Methods for preparing compositions containing the compounds described herein comprise formulating the compounds into solid, semi-solid, or liquid forms with one or more inert, pharmaceutically acceptable excipients or carriers. 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 liposomes, micelles, or nanoparticles containing the compounds disclosed herein. Semi-solid compositions include, but are not limited to, gels, suspensions, and creams. The pharmaceutical compositions described herein may take the form of liquids or suspensions, solids suitable for use in solution or suspension, or emulsions. These compositions may also contain minor amounts of nontoxic auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and the like, as appropriate.

[0125] In some embodiments, the pharmaceutical composition comprising one or more compounds selected from the compound of structure (I) is in a liquid state, for example, the drug is in solution, in suspension, or both.Generally, when the composition is administered as a suspension, the liquid drug is initially in solution, and then in particle form in suspension.In some embodiments, the liquid composition comprises a gel formulation.In other embodiments, the liquid composition is an aqueous solution.

[0126] In some embodiments, aqueous suspensions contain one or more polymers as suspending agents. Polymers include water-soluble polymers (e.g., cellulose polymers such as hydroxypropylmethylcellulose) and water-insoluble polymers (e.g., cross-linked carboxyl-containing polymers). Some pharmaceutical compositions described herein contain a mucoadhesive polymer selected from, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate, and dextran.

[0127] The pharmaceutical compositions also optionally include a solubilizing agent to facilitate dissolution of one or more compounds selected from the compounds of structure (I). The term "solubilizing agent" generally includes agents that form micellar or actual solutions of the compounds. Certain acceptable nonionic surfactants (e.g., polysorbate 80) are useful as solubilizing agents, as are ophthalmically acceptable glycols, polyglycols (e.g., polyethylene glycol 400), and glycol ethers.

[0128] Additionally, pharmaceutical compositions may optionally contain one or more pH adjusters or buffers, including acids (e.g., acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid); bases (e.g., sodium hydroxide, sodium phosphate, borax, sodium citrate, sodium acetate, sodium lactate, and trishydroxymethylaminomethane); and buffers (e.g., citric acid / dextrose, sodium bicarbonate, and ammonium chloride). The above acids, bases, and buffers are included in amounts necessary to maintain the pH of the composition, within acceptable ranges.

[0129] The composition also optionally includes one or more salts in an amount necessary to bring the composition to an acceptable osmolality. 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.

[0130] Other pharmaceutical compositions optionally contain one or more preservatives that inhibit microbial activity. Suitable preservatives include mercury-containing substances (e.g., merfen and thiomersal); stabilized chlorine dioxide; and quaternary ammonium compounds (e.g., benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride).

[0131] 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); and polyoxyethylene alkyl ethers and alkylphenyl ethers (e.g., Octoxynol 10, Octoxynol 40).

[0132] The compositions may also include one or more antioxidants to enhance chemical stability if required. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite.

[0133] In some embodiments, aqueous suspension compositions are packaged in non-resealable single-dose containers, or resealable multi-dose containers are used, in which case the compositions typically contain a preservative.

[0134] In other embodiments, other delivery systems are utilized for hydrophobic pharmaceutical compounds. Liposomes and emulsions are examples of delivery vehicles or carriers useful herein. In certain embodiments, organic solvents (e.g., N-methylpyrrolidone) are also employed. In other embodiments, the compounds described herein are delivered as sustained-release formulations (e.g., semipermeable matrices of solid hydrophobic polymers containing a therapeutic agent). A variety of sustained-release materials are useful herein. In some embodiments, sustained-release capsules release the compound for several weeks up to 100 days or more. Depending on the chemical nature and biological stability of the therapeutic agent, other methods for protein stabilization are employed.

[0135] In certain embodiments, the formulations described herein include one or more antioxidants, metal chelators, thiol-containing compounds, and / or other general stabilizing agents. Examples of such stabilizers include, but are not limited to, (a) about 0.5% to about 2% w / v glycerol, (b) about 0.1% to about 1% w / v methionine, (c) about 0.1% to about 2% w / v monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% to about 2% w / v ascorbic acid, (f) 0.003% to about 0.02% w / v polysorbate 80, (g) 0.001% to about 0.05% w / v polysorbate 20, (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrin, (l) pentosan polysulfate and other heparinoids, (m) divalent cations (e.g., magnesium and zinc); or (n) combinations thereof.

[0136] In some embodiments, the concentration of one or more compounds selected from compounds of structure (I) included in the pharmaceutical compositions of the present disclosure is 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%, 1.25%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.0 Higher than 6%, 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.

[0137] In some embodiments, the pharmaceutical compositions of the present disclosure include one or more compounds selected from the compounds of structure (I) at a concentration of approximately 0.0001% to 50%, approximately 0.001% to 40%, approximately 0.01% to 30%, approximately 0.02% to 29%, approximately 0.03% to 28%, approximately 0.04% to 27%, approximately 0.05% to 26%, approximately 0.06% to 25%, approximately 0.07% ~24%, approximately 0.08%-23%, approximately 0.09%-22%, approximately 0.1%-21%, approximately 0.2%-20%, approximately 0.3%-19%, approximately 0.4%-18%, approximately 0.5%-17%, approximately 0.6%-16%, approximately 0.7%-15%, approximately 0.8%-14%, approximately 0.9%-12%, approximately 1%-10% w / w, w / v or v / v range.

[0138]

[0023] In some embodiments, the amount of one or more compounds selected from the compounds of structure (I) included in the pharmaceutical compositions of the present disclosure is 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0 g, 1 ... 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.

[0139] In some embodiments, the amount of one or more compounds selected from the compounds of structure (I) included in the pharmaceutical compositions of the present disclosure ranges from 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.

[0140] Packaging materials for use in packaging the pharmaceutical compositions described herein include, for example, those described 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 material appropriate for the selected formulation and intended mode of administration and treatment. For example, these containers contain one or more compounds described herein, optionally as a composition or in combination with another agent disclosed herein. Optionally, the container has a sterile access port (e.g., the container is a bag or vial containing an intravenous solution with a stopper pierceable by a hypodermic injection needle). Such kits contain the compounds and, optionally, labeling, labels, or instructions for use in the methods described herein.

[0141] For example, a kit typically includes one or more separate containers, each containing one or more of various items (e.g., appropriately concentrated reagents and / or devices) desired by the supplier and user for use of the compounds described herein. Examples include, but are not limited to, buffers, diluents, filters, needles, syringes; labels on carriers, packages, containers, vials, and / or tubes that describe the contents and / or instructions for use; and package inserts that describe the instructions for use. A set of instructions is usually also included. Optionally, the label is on or associated with the container. For example, a label is on a container when letters, numbers, or other symbols that make up the label are attached, molded, or printed on the container itself, and a label is associated with a container (e.g., as a package insert) when they are inside a container or carrier that holds the container. Furthermore, a label is used to indicate that the contents are used for a specific therapeutic application. Furthermore, the label displays instructions for using the contents, such as the methods described herein. In some embodiments, the pharmaceutical composition is present in a pack or dispensing device containing one or more unit dosage forms having a compound provided herein. The pack may comprise, for example, metal or plastic foil, such as a blister pack. Alternatively, the pack or dispensing device may be accompanied by instructions for administration. Alternatively, the pack or dispensing device may have a notice associated with the container in a format prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, reflecting the agency's approval of the drug form for administration to humans or animals. Such notice may be present, for example, as an approved labeling or approved package insert by the U.S. Food and Drug Administration for prescription drugs. In some embodiments, a composition comprising a compound provided herein formulated in a compatible pharmaceutical carrier is prepared, stored in an appropriate container, and labeled for processing under the intended conditions.

[0142] (method) Embodiments of the present disclosure are useful as modulators of the NLRP3 inflammasome by inhibiting NEK7 in a host. Thus, compounds of structure (I) are also useful in treating conditions mediated by effector signaling molecules such as IL-1β and IL-18.

[0143] The host or patient can be any mammal (e.g., primates, particularly humans; rodents, such as mice, rats, and hamsters; rabbits, horses, cows, dogs, cats, etc.) Animal models provide therapeutic models for human diseases and are useful for experimental investigations.

[0144] In certain embodiments, the present disclosure is useful as an inhibitor of the NLRP3 inflammasome activation mechanism. Thus, the compounds of structure (I) are also useful in treating conditions caused by its activation in a host.

[0145] In another embodiment, the compounds of structure (I) are useful as inhibitors of the NLRP3 (protein)-NEK7 (protein) interaction, and therefore are also useful in treating conditions caused by NLRP3-NEK7 association in a host.

[0146] In certain embodiments, the compounds of structure (I) are useful in the treatment of human conditions mediated by effectors selected from the group consisting of IL-1β, IL-18, and caspase-1.

[0147]

[0010] Embodiments of the present disclosure also relate to the use of a compound according to structure (I) and / or a physiologically acceptable salt thereof for the preventive or therapeutic treatment and / or monitoring of diseases caused by, mediated by, and / or modulated by NLRP3 inflammasome activity. Furthermore, embodiments of the present disclosure relate to the use of a compound according to structure (I) and / or a physiologically acceptable salt thereof in the manufacture of a medicament for the preventive or therapeutic treatment and / or monitoring of diseases caused by, mediated by, and / or modulated by NLRP3 inflammasome activity. In certain embodiments, the present disclosure provides the use of a compound according to structure (I) or a physiologically acceptable salt thereof for the manufacture of a medicament for preventing or treating an NLRP3-mediated disease.

[0148] In another embodiment, the present disclosure relates to a method of treating an inflammatory disease or condition mediated by the NLRP3 inflammasome by administering to a patient in need of treatment a therapeutically effective amount of a compound of structure (I).

[0149] In certain embodiments, diseases that may be treated with the compounds of structure (I) include type 2 diabetes, atherosclerosis, Alzheimer's disease, aging, fatty liver, metabolic syndrome, asthma, psoriasis, obesity, acute and chronic tissue damage due to infection, gout, arthritis, enteritis, hepatitis, peritonitis, silicosis, UV-induced skin sunburn, contact hypersensitivity, sepsis, cancer, neurodegenerative diseases, multiple sclerosis, Muckle-Wells syndrome, and combinations thereof.

[0150] In certain other embodiments, the compounds of structure (I) are used in a method for treating a disorder or disease selected from autoimmune diseases, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, bacterial and viral infections, allergies, asthma, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, transplantation, sperm motility disorders, red blood cell deficiencies, graft rejection, pulmonary disorders, respiratory diseases, ischemic conditions, cancer, and combinations thereof.

[0151] In some embodiments, the NEK7-associated disease treatable with a compound of structure (I) is 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), hyper-IgD and periodic fever syndrome, cryopyrin-associated periodic fever syndrome, Schnitzler's syndrome, systemic juvenile idiopathic arthritis, adult-onset Still's disease, gout, pseudogout, SAPHO syndrome, Castleman's disease, sepsis, stroke, atherosclerosis, celiac disease, DIRA (IL-1 receptor antagonist molecule deficiency), Alzheimer's disease, Parkinson's disease, cancer, and combinations thereof.

[0152] Also included herein are methods of treatment in which at least one compound of structure (I) is administered in combination with an anti-inflammatory or therapeutic agent. Anti-inflammatory agents include, but are not limited to, NSAIDs, nonspecific and COX-2 nonspecific and specific 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, a combination of diclofenac sodium and misoprostol, sulindac, oxaprozin, diflunisal, piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen, ketoprofen sodium, nabumetone, sulfasalazine, tolmetin sodium, hydroxychloroquine, and combinations thereof.

[0153] Examples of NSAIDs also include COX-2 specific inhibitors (eg, celecoxib, valdecoxib, lumiracoxib, and / or etoricoxib).

[0154] In some embodiments, the anti-inflammatory agent is a salicylate, including but not limited to acetylsalicylic acid or aspirin, sodium salicylate, and choline and magnesium salicylates.

[0155] The anti-inflammatory agent can also be a corticosteroid, for example, the corticosteroid can be cortisone, dexamethasone, methylprednisolone, prednisolone, prednisolone sodium phosphate, or prednisone.

[0156] In another embodiment, the anti-inflammatory agent and a gold compound (eg, sodium aurothiomalate or auranofin).

[0157] The present disclosure also includes embodiments in which the anti-inflammatory agent is a metabolic inhibitor (e.g., a dihydrofolate reductase inhibitor such as methotrexate, or a dihydrofolate dehydrogenase inhibitor such as leflunomide).

[0158] Therapeutic agents may also include agents for pain and inflammation such as, for example, histamine and antihistamines, bradykinin and bradykinin receptor antagonists, 5-hydroxytryptamine (serotonin), lipid substances produced by the biotransformation of selective hydrolysis products of membrane phospholipids, eicosanoids, prostaglandins, thromboxanes, leukotrienes, aspirin, nonsteroidal anti-inflammatory drugs, antipyretics, analgesics, agents that inhibit the synthesis of prostaglandins and thromboxanes, selective inhibitors of inducible cyclooxygenase, selective inhibitors of inducible cyclooxygenase-2, autacoids, paracrine hormones, somatostatin, gastrin, cytokines that mediate interactions involved in humoral and cellular immune responses, lipid-derived autacoids, eicosanoids, β-adrenergic receptor agonists, ipratropium, glucocorticoids, methylxanthines, sodium channel blockers, opioid receptor agonists, calcium channel blockers, membrane stabilizers, and leukotriene inhibitors.

[0159] 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 pexelizumab), a TNF antagonist (e.g., etanercept, or the anti-TNFα monoclonal antibody infliximab).

[0160] Therapeutic agents used in combination with compounds of structure (I) can also include small molecule compounds that inhibit NLRP3 inflammasome activation (e.g., MCC950, sulforaphane, isoliquiritigenin, β-hydroxybutyric acid, flufenamic acid, mefenamic acid, 3,4-methylenedioxy-β-nitrostyrene (MNS), and parthenolide).

[0161] Yet other embodiments of the present disclosure relate to combinations in which at least one active agent is an immunosuppressant compound (e.g., an immunosuppressant compound selected from methotrexate, leflunomide, cyclosporine, tacrolimus, azathioprine, and mycophenolate mofetil).

[0162] The disclosed compounds of structure (I) can be administered in combination with other known therapeutic agents, including anti-cancer agents. As used herein, the term "anti-cancer agent" refers to any agent administered to a cancer patient for the purpose of treating the cancer.

[0163] In some embodiments, the anti-cancer agents are classified into the following categories: Alkylating agents: e.g., altretamine, bendamustine, busulfan, carmustine, chlorambucil, chlormethine, cyclophosphamide, dacarbazine, ifosfamide, improsulfan, tosylate, lomustine, melphalan, mitobronitol, mitolactol, nimustine, ranimustine, temozolomide, thiotepa, treosulfan, mechlorethamine, carboquone, apaziquone, fotemustine, glufosfamide, palifosfamide, pipobroman, trofosfamide, uramustine, TH-3024, VAL-0834; Platinum compounds: e.g., carboplatin, cisplatin, eptaplatin, miriplatin hydrate, oxaliplatin, lobaplatin, nedaplatin, picoplatin, satraplatin, lobaplatin, nedaplatin, picoplatin, satraplatin; DNA mutagenic agents: e.g., amrubicin, bisantrene, decitabine, mitoxantrone, procarbazine, trabectedin, clofarabine, amsacrine, brostallicin, pixantrone, laromustine; Topoisomerase inhibitors: e.g. etoposide, irinotecan, razoxane, sobuzoxane, teniposide, topotecan, amonafide, belotecan, elliptinium acetate, voreloxine; Microtubule inhibitors: e.g., cabazitaxel, docetaxel, eribulin, ixabepilone, paclitaxel, vinblastine, vincristine, vinorelbine, vindesine, vinflunine, fosbretabine, tesetaxel; Antimetabolites: e.g., asparaginase 3, azacitidine, levofolinate calcium, capecitabine, cladribine, cytarabine, enocitabine, floxuridine, fludarabine, fluorouracil, gemcitabine, mercaptopurine, methotrexate, nelarabine, pemetrexed, pralatrexate, azathioprine, thioguanine, carmofur, doxifluridine, elacitarabine, raltitrexed, sapacitabine, tegafur, trimetrexate; Anticancer antibiotics: for example, bleomycin, dactinomycin, doxorubicin, epirubicin, idarubicin, levamisole, miltefosine, mitomycin C, romidepsin, streptozocin, valrubicin, zinostatin, zorubicin, daunorubicin, plicamycin, aclarubicin, peplomycin, pirarubicin; Hormones / antagonists: e.g., abarelix, abiraterone, bicalutamide, buserelin, calsterone, chlorotrianisene, degarelix, dexamethasone, estradiol, fluocortolone, fluoxymesterone, flutamide, fulvestrant, goserelin, histrelin, leuprorelin, megestrol, mitotane, nafarelin, nandrolone, nilutamide, octreotide, prednisolone, raloxifene, tamoxifen, human thyrotropin alfa, toremifene, trilostane, triptorelin, diethylstilbestrol, acolbifene, danazol, deslorelin, epithiostanol, orteronel, enzalutamide; Aromatase inhibitors: e.g., aminoglutethimide, anastrozole, exemestane, fadrozole, letrozole, testolactone, formestane; Small molecule kinase inhibitors: e.g., crizotinib, dasatinib, erlotinib, imatinib, lapatinib, nilotinib, pazopanib, regorafenib, ruxolitinib, sorafenib, sunitinib, vandetanib, vemurafenib, bosutinib, gefitinib, axitinib, afatinib, alisertib, dabrafenib, dacomitinib, dinaciclib, dovitinib , enzastaurin, nintedanib, lenvatinib, linifanib, linsitinib, masitinib, midostaurin, motesanib, neratinib, orantinib, perifosine, ponatinib, radotinib, rigosertib, tipifarnib, tivantinib, tivozanib, trametinib, pimasertib, brivanib alanine ester, cediranib

[0164] In some embodiments, the pharmaceuticals administered with the compounds described herein include any suitable drug that can be effectively delivered by inhalation. For example, analgesics (e.g., codeine, dihydromorphine, ergotamine, fentanyl, or morphine); antianginal drugs (e.g., diltiazem); antiallergic drugs (e.g., cromoglycate, ketotifen, or nedocromil); antiinfective drugs (e.g., cephalosporins, penicillin antibiotics, streptomycin, sulfonamides, tetracycline, or pentamidine); antihistamines (e.g., methapyrilene); anti-inflammatory drugs (e.g., beclomethasone, flunisolide, budesonide, tipredane, triamcinolone acetonide, or fluticasone); antitussives (e.g., noscapine); bronchodilators (e.g., ephedrine, adrenaline, fenoterol, formoterol, isoprenaline, metaproterenol, phenylephrine, phenylpropanolamine, pirbuterol, reproterol, rimiterol, salbutamol, salmeterol, terbutaline, isoetharine, 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 oxitropium); hormones (e.g., cortisone, hydrocortisone, or prednisolone); xanthines (e.g., aminophylline, choline theophylline, lysine theophylline) theophyllinate or theophylline); and therapeutic proteins and peptides (e.g., insulin or glucagon). It will be apparent to those skilled in the art that, where appropriate, the drug will be used in the form of a salt (e.g., as an alkali metal or amine salt, or as an acid addition salt), or as an ester (e.g., a lower alkyl ester), or as a solvate (e.g., a hydrate) in order to maximize the activity and / or stability of the drug.

[0165] The drug of the present disclosure or other suitable drug is administered depending on the symptoms to be treated. Thus, in some embodiments, one or more compounds of the present disclosure are administered simultaneously with the other drugs described above. When used in combination therapy, the compound described herein is administered simultaneously with the second drug or separately. This combined administration can include simultaneous administration of the two drugs in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, the compound described herein and any of the above drugs can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compound of the present disclosure and any of the above drugs can be administered simultaneously in separate dosage forms. Alternatively, the compound of the present disclosure can be administered immediately after any of the above drugs, or vice versa. In some embodiments of the separate administration method, the compound of the present disclosure and any of the above drugs are administered within minutes, hours, or days of each other.

[0166] In some embodiments, the compound of structure (I) is administered as a monotherapy.

[0167] To identify signal transduction or mechanistic pathways and discover interactions between various signal transduction pathways, various scientists have developed appropriate models or model systems (e.g., cell culture models and transgenic animal models). Interacting compounds can be used to modulate signals in determining specific steps in the signal transduction cascade. Compounds of embodiments of the present disclosure can also be used as reagents to study NEK7-dependent signal transduction pathways in animal and / or cell culture models, or in clinical diseases mentioned herein.

[0168] The methods of the presently disclosed embodiments can be carried out either in vitro or in vivo. The sensitivity of a particular cell to treatment with a compound of structure (I) can be specifically determined by in vitro testing, either in the research or clinical stages. Generally, cultured cells are treated with various concentrations of the compound for a time sufficient for the active agent to inhibit NEK7 activity, usually from about one hour to one week. In vitro treatment can be carried out using cells cultured from a biopsy sample or a cell line.

[0169] In some embodiments, the IC of a compound of structure (I) for inhibiting NEK7 50 was determined by the concentration of compound required to inhibit NEK kinase activity by 50%. Compounds of structure (I) have an IC50 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 values ​​were shown.

[0170] The following examples and preparations further illustrate and exemplify the compounds of the present disclosure, methods for making and testing the compounds. It should be understood that the scope of the present disclosure is not limited in any way by the following examples and preparations. In the following examples, and throughout the specification and claims, molecules with one stereocenter exist as a racemic mixture unless otherwise specified. Molecules with two or more stereocenters exist as a racemic mixture of diastereomers unless otherwise specified. Single enantiomers / diastereomers can be obtained by methods known to those skilled in the art.

[0171] Methods for preparing the compounds described herein are provided below. Generally, the starting components can be obtained from Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, Fluorochem USA, etc., or can be synthesized according to literature known to those skilled in the art (see, for example, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th edition (Wiley, December 2000)), or can be prepared as described herein.

[0172] The following examples illustrate the structure (I): [ka] (In the formula, each A, X, Y, Z, R 1 , R 2 , R 3 and n is defined herein), or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof.

[0173] Those skilled in the art will also understand that in the preparation of compounds described herein, it may be necessary to protect functional groups of intermediate compounds with suitable protecting groups. Such functional groups include, but are not limited to, hydroxy, amino, mercapto, and carboxy. Suitable protecting groups for hydroxy groups include trialkylsilyl or diarylalkylsilyl (e.g., t-butyldimethylsilyl, t-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino, amidino, and guanidino groups include t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for mercapto groups include -C(O)-R" (where R" is alkyl, aryl, or arylalkyl), p-methoxybenzyl, trityl, and the like. Suitable protecting groups for carboxy groups include alkyl, aryl, or arylalkyl esters. Protecting groups are known to those skilled in the art and may be added or removed as appropriate using standard methods 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. Those skilled in the art will also appreciate that the protecting group can be a polymeric resin (e.g., Wang resin, Rink resin, or 2-chlorotrityl chloride resin).

[0174] It will also be understood by those skilled in the art that protected derivatives of the disclosed compounds may not have pharmacological activity, but may be administered to a mammal and subsequently metabolized in the body to form a pharmacologically active compound of the disclosed compounds. Such derivatives are herein referred to as "prodrugs." Prodrugs of the disclosed compounds are included within the scope of embodiments of the present disclosure.

[0175] Example The following examples are offered by way of illustration.

[0176] General Procedure All proton NMR experiments were performed at 400 MHz on a Bruker NEO spectrometer equipped with a BBFO probe. Deuterated solvents contained less than 0.05% v / v tetramethylsilane, which was used as the reference signal (set at 0.00 ppm). When the deuterated solvent did not contain tetramethylsilane, the remaining non-deuterated solvent peak was used as the reference signal, following published guidelines (J. Org. Chem. 1997, 62(21), 7512-7515). Chemical shifts are expressed in parts per million (ppm, δ units). Coupling constants are expressed in hertz (Hz). Fragmentation patterns indicate apparent multiplicity and are designated as s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), qt (quintet), or brs (broad signal). LC / MS analysis was performed on an Agilent Technologies UHPLC 1290 Infinity II and G6125 MS detector. Microwave reactions were carried out using a Monowave 300 (Anton Paar GmbH) according to standard protocols.

[0177] NEK7 enzyme assay Casein substrate (a mixture of hydrolyzed and partially dephosphorylated α-, β-, and κ-caseins from bovine milk, purchased from Sigma-Aldrich, catalog number C4765, diluted in distilled water to a final concentration of 1 mg / mL) and full-length recombinant human NEK7 (expressed by baculovirus in Sf9 insect cells with an N-terminal GST tag, purchased from SignalChem, catalog number 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). Compounds of interest (3-fold serial dilutions from 10 μM to 0.5 nM in DMSO) or vehicle (1% DMSO) were dispensed into the kinase reaction mixture using sonic technology (Echo550, nanoliter range). After 20 min of incubation at room temperature, 32The kinase reaction was initiated by adding [P]-ATP (specific activity at 10 μCi / μL), and the mixture was incubated at room temperature for 2 hours. The reaction was then stopped by spotting the reaction mixture onto phosphocellulose paper (P81). After washing, the specific radioactivity of the P81 paper was measured, and kinase activity data were expressed as the percentage of kinase activity in the test sample compared to the vehicle reaction. IC 50 Value calculation and curve fitting were performed using Prism (GraphPad Software).

[0178] IL-1β release assay Approximately 1.5 million THP-1 cells were seeded into each well of a 6-well TC plate and incubated with 40 nM PMA / RPMI (10% FBS, 1% Pen-Strep) for 24 hours. The medium was then removed, and the cells were rested in RPMI (10% FBS, 1% Pen-Strep) for 24 hours. After this, the medium was removed, and the cells were pre-treated with various concentrations of the target compound (typically 3-fold serial dilutions in RPMI + 5% FBS, concentrations: 1 μM to 0.5 nM) in RPMI (5% FBS) for 2 hours. The medium was removed again, and the cells were incubated with LPS (250 ng / mL) and the target compound (at the concentrations listed above) in RPMI (5% FBS) for 2 hours. Finally, the medium was removed, and the cells were incubated with nigericin (20 μM) and the target compound (at the concentrations listed above) in Opti-MEM for 30 minutes. The cell culture medium was then collected and the amount of bound IL-1β was determined using JESS (Protein Simple) and standard protocols. The bound IL-1β antibody was obtained from Cell Signaling (Cat. No. 83186S) and diluted 1:20 with Antibody Diluent 2. A 1x anti-rabbit HRP secondary antibody (Protein Simple) was used for chemiluminescence detection with Protein Simple's luminol and peroxide. Incubation time with the primary antibody was increased from 30 to 60 minutes.

[0179] Abbreviation °C (degrees Celsius); 1H NMR (proton nuclear magnetic resonance); ACN (acetonitrile); AIBN (azobisisobutyronitrile); bipy (2,2'-bipyridine); (Boc)2O (di-tert-butyl dicarbonate); Cu(OAC)2 (copper(II) acetate); DCE (dichloroethane); DCM (dichloromethane); DIPEA (N,N-diisopropylethylamine); DMAP (4-dimethylaminopyridine); DMF (N,N-dimethylformamide); DMSO-d6 (deuterated dimethyl sulfoxide); eq. (equivalent); EtOAc (ethyl acetate); g (gram); h (hour); HPLC (high performance liquid chromatography); LC-MS (liquid chromatography mass spectrometry); MeOH (methanol); mg (milligram); min (minute); mL (milliliter); mmol (millimol); MsCl (methanesulfonyl chloride); NBS (N-bromosuccinimide); NIS (N-iodosuccinimide); Pd2(dba)3 (tris(dibenzylideneacetone)dipalladium(0)); Pd(PPh3)4 (palladium-tetrakis(triphenylphosphine)); PdCl2(dppf) ([1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II)); STAB (sodium triacetoxyborohydride); TBAF (tetra-n-butylammonium fluoride); TEA (triethylamine); TFA (trifluoroacetic acid); THF (tetrahydrofuran); TLC (thin layer chromatography); XPhos (2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl)

[0180] Production of synthetic intermediates Intermediate A1 1-Cyclopropyl-3-iodo-1H-pyrazolo[3,4-D]pyrimidin-4-amine [ka] Cu(OAc) (0.348 g, 1.916 mmol), bipy (0.299 g, 1.916 mmol), and NaHCO (0.322 g, 3.830 mmol) were added to a stirred solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.500 g, 1.916 mmol) and cyclopropylboronic acid (0.329 g, 3.830 mmol) in dichloroethane (10 mL) and stirred at 70 °C under an oxygen atmosphere for 12 hours. After completion of the reaction (as confirmed by TLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), which was then washed with DCM (2 × 20 mL). The combined filtrate was washed with water (20 mL) and brine (25 mL), and the organic layer was separated, dried over NaSO, filtered, and concentrated under reduced pressure to provide the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), elution: 20% EtOAc / petroleum ether) to give the title compound as an off-white solid (0.24 g, 36% yield). 1 LCMS: 301.8 [M+H]

[0181] Intermediate A2 3-Iodo-1-isopropyl-1H-pyrazolo[3,4-D]pyrimidin-4-amine [ka] CsCO (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]pyrimidin-4-amine (5.00 g, 19.15 mmol) in DMF (25 mL), and the mixture was stirred at 90 °C for 16 h in a sealed tube. After completion of the reaction (as determined by TLC), it was poured onto crushed ice (50 g) and stirred for 15 min. The resulting solid was filtered, washed with water (2 × 5 mL), and dried to give the title compound as an off-white solid (3.25 g, 56% yield). 1H NMR (400MHz, DMSO-d6)δ=8.18(s, 1H), 4.93-4.99(m, 1H), 1.42(d, J=6.8Hz, 6H); LCMS: 303.8 [M+H]

[0182] Intermediate A3 3-Iodo-1-(oxetan-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-4-amine [ka] The title compound was prepared in a similar manner to that described for Intermediate A2, substituting 3-iodooxetane for 2-iodopropane. 1 H NMR (400MHz, DMSO-d6)δ=8.21(s, 1H), 5.89-5.93(m, 1H), 4.93-5.00(m, 4H); LCMS: 318.0 [M+H]

[0183] Intermediate A4 1-(4-amino-3-iodo-1H-pyrazolo[3,4-D]pyrimidin-1-yl)-2-methylpropan-2-ol [ka] NaH2PO4 (0.044 g, 0.372 mmol) was added to a mixture of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-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), and the resulting solution was microwaved at 150 °C for 1 h. After completion of the reaction (as confirmed by TLC), the solvent was removed under reduced pressure to give the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), eluting with 25% EtOAc / petroleum ether) to give the title compound as a light brown solid (0.064 g, 51% yield). 1H NMR (400MHz, DMSO-d6)δ=8.20(s, 1H), 4.19(s, 2H), 1.09(s, 6H); LCMS: 334.0 [M+H]

[0184] Intermediate A5 7-Cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0185] Step 1: Synthesis of 4-chloro-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidine [ka] NaHCO (10.94 g, 130.0 mmol), bipy (10.17 g, 65.1 mmol), and Cu(OAc) (11.83 g, 65.1 mmol) were added to a solution of 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (10.00 g, 65.1 mmol) and cyclopropylboronic acid (11.19 g, 130.0 mmol) in DCE (150 mL), and the resulting mixture was stirred at 80 °C for 12 h. After completion of the reaction (as confirmed by TLC), the reaction mixture was filtered through Celite (i.e., diatomaceous earth), which was then washed with DCM (2 × 100 mL). The combined filtrate was washed with water (100 mL) and brine (100 mL), dried over NaSO, filtered, and concentrated under reduced pressure to provide the crude material. This was purified by GRACE (silica gel (230-400 mesh), elution: 6% EtOAc / petroleum ether) to give the title compound as an off-white solid (7.30 g, yield 58%). 1 H NMR (400MHz, DMSO-d6)δ=8.65(s, 1H), 7.70(d, J=3.6Hz, 1H), 6.59(d, J=4.0Hz, 1H), 3.62-3.68(m, 1H), 1.08-1.10(m, 4H); LCMS: 194.0 [M+H]

[0186] Step 2: Synthesis of 4-chloro-7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidine [ka] To a solution of 4-chloro-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidine (7.10 g, 36.7 mmol) in DMF (50 mL) was added NIS (8.25 g, 36.7 mmol) at 0° C., and the resulting mixture was stirred at 25° C. for 12 hours. After completion of the reaction (confirmed by TLC), the reaction mixture was added to ice-cold water (600 mL) and stirred at 25° C. for 15 minutes. The resulting precipitate was filtered, washed with water (2×250 mL), and dried under reduced pressure to give the title product (8.30 g, 70% yield) as a light brown solid. 1 H NMR (400MHz, DMSO-d6)δ=8.65(s, 1H), 7.94(s, 1H), 3.62-3.68(m, 1H), 1.05-1.10(m, 1H); LCMS: 319.9 [M+H]

[0187] Step 3: Synthesis of 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] A mixture of 4-chloro-7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (8.3 g, 26 mmol) and ammonia (25% aqueous solution, 25 mL) in 1,4-dioxane (25 mL) was loaded into a tinyclave and stirred for 16 h at 120° C. After completion of the reaction (as confirmed by TLC), the reaction mixture was cooled to 25° C. and the resulting precipitate was filtered, washed with petroleum ether (2×50 mL), and dried to give the title compound as an off-white solid (6.0 g, 76% yield). 1 H NMR (400MHz, DMSO-d6)δ=8.11(s, 1H), 7.38(s, 1H), 6.59(bs, 2H), 3.48-3.54(m, 1H), 0.97-0.99(m, 4H); LCMS: 300.9 [M+H]

[0188] Intermediate A6 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0189] Step 1: Synthesis of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine [ka] To a solution of 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (5.00 g, 32.6 mmol) in DMF (20 mL) was added NIS (7.33 g, 32.6 mmol) at 0° C., and the resulting mixture was stirred at 25° C. for 12 hours. After completion of the reaction (as confirmed by TLC), the reaction mixture was added to crushed ice (200 g) and stirred at 25° C. for 30 minutes. The resulting precipitate was filtered, washed with water (2×20 mL), and dried to give the title compound as a light brown solid (8.52 g, 92% yield). 1 H NMR (400MHz, DMSO-d6)δ=12.96(bs, 1H), 8.60(s, 1H), 7.94(s, 1H); LCMS: 280.0 [M+H]

[0190] Step 2: Synthesis of 4-chloro-5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine [ka] A solution of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (0.500 g, 1.789 mmol) in THF (5 mL) was added dropwise to a stirred suspension of NaH (60%, dispersed in mineral oil, 0.143 g, 3.580 mmol) in THF (15 mL) at 0 °C, and the resulting mixture was stirred at 0 °C for 30 min. Iodomethane (0.134 mL, 2.147 mmol) was then added, and the resulting mixture was stirred at 25 °C for 4 h. After completion of the reaction (confirmed by TLC), the reaction mixture was quenched with water (10 mL) and extracted with EtOAc (2 × 10 mL). The combined organic extracts were dried over NaSO, filtered, and concentrated under reduced pressure to give the title compound as an off-white solid (0.38 g, 71% yield). LCMS: 294.0 [M+H]

[0191] Step 3: Synthesis of 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] A mixture of 4-chloro-5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine (0.40 g, 1.363 mmol) and ammonia (25% aqueous solution, 4 mL) in 1,4-dioxane (4 mL) was subjected to microwave irradiation at 150 °C for 1 h. After completion of the reaction (as confirmed by LCMS), the reaction mixture was concentrated under reduced pressure to give the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), elution: 10% MeOH / DCM) to give the title compound as an off-white solid (0.12 g, 32% yield). 1 H NMR (400MHz, DMSO-d6)δ=8.10(s, 1H), 7.42(s, 1H), 6.59(bs, 2H), 3.67(s, 3H); LCMS: 275.0 [M+H]

[0192] Intermediate A7 7-Cyclobutyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0193] Step 1: Synthesis of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine [ka] Bromocyclobutane (0.580 g, 4.29 mmol) and K2CO3 (0.742 g, 5.37 mmol) were added to a solution of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (1.000 g, 3.58 mmol) in DMF (5 mL), and the resulting mixture was stirred at 80 °C for 16 h. After completion of the reaction (as confirmed by TLC), the reaction mixture was added to crushed ice (25 g). The resulting precipitate was filtered, washed with water (20 mL), and dried to give the crude material. This was purified by Isolera (silica gel (230-400 mesh), elution: 20% EtOAc / petroleum) to give the title compound as an off-white solid (0.35 g, 29% yield). LCMS: 333.9 [M+H]

[0194] Step 2: Synthesis of 7-cyclobutyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] A mixture of 4-chloro-7-cyclobutyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (0.35 g, 1.049 mmol) and ammonia (25% aqueous solution, 4.5 mL) in 1,4-dioxane (4.5 mL) was microwaved at 150 °C for 1 hour. After completion of the reaction (as confirmed by TLC), the reaction mixture was concentrated under reduced pressure to give the title compound as an off-white solid (0.30 g, 83% yield), which was used without further purification. 1 H NMR (400MHz, DMSO-d6)δ=8.09(s, 1H), 7.73(s, 1H), 6.61(bs, 2H), 5.09-5.16(m, 1H), 2.50-2.50(m, 2H), 2.31-2.38(m, 2H), 1.75-1.83(m, 2H); LCMS: 315.0 [M+H]

[0195] Intermediate A8 1-(4-amino-5-iodo-7H-pyrrolo[2,3-D]pyrimidin-7-yl)-2-methylpropan-2-ol [ka] The title compound was prepared as described in PCT International Publication No. WO 2021 / 226547 A2. Intermediate A9 5-Iodo-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-D]pyrimidin-4-amine [ka] The title compound was prepared as described in PCT International Publication No. WO 2014 / 184069 A1.

[0196] Intermediate A10 5-Iodo-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-D]pyrimidin-4-amine [ka] The title compound was prepared as described in PCT International Publication No. WO 2017 / 220477 A1.

[0197] Intermediate A11 1-Cyclopropyl-N-(2,4-dimethoxybenzyl)-3-iodo-1H-pyrazolo[4,3-c]pyridin-4-amine

[0198] Step 1: Synthesis of 4-chloro-3-iodo-1H-pyrazolo[4,3-c]pyridine [ka] KOH (1.32 g, 23.0 mmol) and iodine (1.62 g, 12.8 mmol) were added to a solution of 4-chloro-1H-pyrazolo[4,3-c]pyridine (1.00 g, 6.4 mmol) in 1,4-dioxane (10 mL), and the resulting mixture was stirred at 75° C. for 4 hours. After completion of the reaction (as confirmed by TLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth). The resulting filtrate was concentrated under reduced pressure to give the crude material. This was purified by reverse-phase column chromatography to give the title compound as a white solid (0.633 g, 63% yield). 1 H NMR(400MHz, DMSO-d6)δ=14.12(bs, 1H), 8.14(d, J=6.0Hz, 1H), 7.66(d, J=5.6Hz, 1H); LCMS: 279.9 [M+H]

[0199] Step 2: Synthesis of 4-chloro-1-cyclopropyl-3-iodo-1H-pyrazolo[4,3-c]pyridine [ka] The title compound was prepared in a similar manner as described in Intermediate A1, starting from 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), and obtained as a white solid (0.430 g, 60% yield). 1 H NMR (400MHz, DMSO-d6)δ=8.21(d, J=6.0Hz, 1H), 7.81(d, J=5.6Hz, 1H), 3.84-3.89(m, 1H), 1.14-1.17(m, 4H); LCMS: 319.7 [M+H]

[0200] Step 3: Synthesis of 1-cyclopropyl-N-(2,4-dimethoxybenzyl)-3-iodo-1H-pyrazolo[4,3-c]pyridin-4-amine [ka] A mixture of 4-chloro-1-cyclopropyl-3-iodo-1H-pyrazolo[4,3-c]pyridine (0.880 g, 2.75 mmol) and (2,4-dimethoxyphenyl)methanamine (1.245 mL, 8.26 mmol) in nBuOH (10 mL) was stirred at 110 °C for 12 hours. After completion of the reaction (confirmed by LCMS), the reaction mixture was concentrated under reduced pressure to give the crude material. This was purified by Isolera (silica gel (230-400 mesh), elution: 40% EtOAc / petroleum ether) to give the title product as a yellow sticky material (1.0 g, 80% yield). 1 H NMR (400MHz, DMSO-d6)δ=7.81(d, J=6.4Hz, 1H), 7.19(d, J=8.4Hz, 1H), 6.87(d, J=6.0Hz, 1H), 6.60(d, J=2.4Hz, 1H), 6.53-6.56(m, 1H), 6.45-6.47(m, 1H), 4.62(d, J=5.6Hz, 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]

[0201] Intermediate A12 2-(4-amino-3-iodo-1H-pyrazolo[3,4-D]pyrimidin-1-yl)ethan-1-ol [ka] The title compound was prepared as described in PCT International Publication No. WO 2011 / 119663 A1.

[0202] Intermediate A13 3-Iodo-1-(2-methoxyethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine [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]pyrimidin-4-amine (0.500 g, 1.916 mmol) in DMF (6 mL), and the resulting solution was stirred at 80 °C for 12 h in a sealed tube. After completion of the reaction (as confirmed by TLC), the reaction mixture was added to crushed ice (25 g) and extracted with EtOAc (2 × 50 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure to give the title product (0.500 g, crude), which was used without further purification. 1 LCMS: 319.8 [M+H]

[0203] Intermediate A14 5-Iodo-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] The title compound was prepared as described in PCT International Publication No. WO 2014 / 184069 A1.

[0204] Intermediate A15 5-Iodo-7-(1-(methylsulfonyl)piperidin-4-yl)-7H-pyrrolo[2,3-D]pyrimidin-4-amine

[0205] Step 1: Synthesis of tert-butyl 4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate [ka] Cs2CO3 (3.76 g, 11.54 mmol) and tert-butyl 4-(tosyloxy)piperidine-1-carboxylate (synthesized as described in PCT International Publication No. WO 2018 / 050771 A1, 4.10 g, 11.54 mmol) were added to a solution of 5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (1.50 g, 5.77 mmol) in DMF (15 mL), and the resulting mixture was stirred at 90 °C for 12 h. After completion of the reaction (as confirmed by LCMS), the reaction mixture was added to ice-cold water (100 mL) and extracted with EtOAc (3 × 50 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), elution: 70% EtOAc / petroleum ether) to give the title compound as a yellow solid (1.2 g, 47% yield). LCMS: 444.1 [M+H]

[0206] Step 2: Synthesis of 5-iodo-7-(piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] HCl / 1,4-dioxane (4M, 6 mL) was added to a solution of tert-butyl 4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate (1.20 g, 2.71 mmol) in DCM (15 mL) at 0° C., and the resulting mixture was stirred at 25° C. for 16 hours. After completion of the reaction (as confirmed by LCMS), the reaction mixture was concentrated under reduced pressure to give the title compound (0.697 g, crude) as an off-white solid, which was used without further purification. LCMS: 344.1 [M+H]

[0207] Step 3: Synthesis of 5-iodo-7-(1-(methylsulfonyl)piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] TEA (0.205 mL, 1.457 mmol) and MsCl (0.057 mL, 0.728 mmol) were added to a solution of 5-iodo-7-(piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (0.250 g, 0.728 mmol) in DCM (5 mL) at 0 °C, and the resulting mixture was stirred at 25 °C for 16 h. After completion of the reaction (confirmed by TLC), the reaction mixture was diluted with DCM (10 mL) and washed with brine (2 mL). The organic layer was dried over Na SO , filtered, and concentrated under reduced pressure to give the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), elution: 10% MeOH / DCM) to give the title compound as a light brown solid (0.28 g, 91% yield). LCMS: 422.0 [M+H]

[0208] Intermediate A16 7-(1-(cyclopropylsulfonyl)piperidin-4-yl)-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] The title compound was prepared in a manner similar to that described in Step 3 of Intermediate A15, starting from 5-iodo-7-(piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (0.300 g, 0.874 mmol) and cyclopropanesulfonyl chloride (0.123 g, 0.874 mmol), and obtained as a pale brown sticky material (0.101 g, 46% yield). LCMS: 448.0 [M+H]

[0209] Intermediate A17 5-(4-amino-3-iodo-1H-pyrazolo[3,4-D]pyrimidin-1-yl)pentan-1-ol [ka] CsCO (0.187 g, 0.383 mmol) and 5-bromopentan-1-ol (0.064 g, 0.383 mmol) were added to a solution of 5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (0.100 g, 0.383 mmol) in DMF (2 mL), and the resulting mixture was stirred at 80 °C for 12 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was added to ice-cold water (100 mL) and extracted with EtOAc (3 × 10 mL). The combined organic extracts were dried over NaSO, filtered, and concentrated under reduced pressure to give the title compound as a light brown sticky material (0.130 g, crude), which was used without further purification. LCMS: 348.0 [M+H]

[0210] Intermediate A18 5-Iodo-7-(1-methylazetidin-3-yl)-7H-pyrrolo[2,3-D]pyrimidin-4-amine

[0211] Step 1: Synthesis of 7-(azetidin-3-yl)-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] TFA (2.5 mL) was added to a solution of tert-butyl 3-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)azetidine-1-carboxylate (synthesized as described in PCT International Publication No. WO 2017 / 215485 A1, 0.520 g, 1.252 mmol) in DCM (5 mL), and the mixture was stirred at 25° C. for 12 hours. After completion of the reaction (confirmed by UPLC), the reaction mixture was concentrated under reduced pressure to give the title compound as an off-white solid (0.4 g, crude), which was used without further purification. LCMS: 316.0 [M+H]

[0212] Step 2: Synthesis of 5-iodo-7-(1-methylazetidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] Paraformaldehyde (0.149 g, 4.950 mmol) and acetic acid (4.95 mg, 0.083 mmol) were added to a solution of 7-(azetidin-3-yl)-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (0.520 g, 1.650 mmol) in MeOH (5 mL), and the mixture was stirred at 50° C. for 2 hours. NaBH3CN (0.518 g, 8.25 mmol) was then added, and the reaction mixture was stirred at 50° C. for an additional 12 hours. After completion of the reaction (confirmed by LCMS), the reaction mixture was concentrated under reduced pressure to give the title compound as an off-white solid (0.30 g, crude), which was used without further purification. LCMS: 330.0 [M+H]

[0213] Intermediate A19 5-Iodo-7-isopropyl-7H-pyrrolo[2,3-D]pyrimidin-4-amine [ka] The title compound was prepared as described in PCT International Publication No. WO 2017 / 220477 A1.

[0214] Intermediate A20 tert-Butyl 3-(4-amino-5-iodo-7H-pyrrolo[2,3-D]pyrimidin-7-yl)propanoate [ka] CsCO (2.506 g, 7.69 mmol) and tert-butyl 3-bromopropanoate (0.804 g, 3.85 mmol) were added to a solution of 5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (1.000 g, 3.85 mmol) in DMF (5 mL), and the mixture was stirred at 90 °C for 12 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was added to crushed ice (25 g) and extracted with EtOAc (3 × 25 mL). The combined filtrate was dried over NaSO, filtered, and concentrated under reduced pressure to give the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), elution: 40% EtOAc / petroleum ether) to give the title compound as a light brown solid (0.34 g, 23% yield). LCMS: 389.0 [M+H]

[0215] Intermediate A21 tert-Butyl 4-(4-amino-5-iodo-7H-pyrrolo[2,3-D]pyrimidin-7-yl)butanoate [ka] The title compound was prepared in a similar manner as described in Intermediate A20, starting from 5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (1.000 g, 3.85 mmol) and tert-butyl 4-bromobutanoate (0.858 g, 3.85 mmol), and obtained as a brown solid (1.28 g, 83% yield). LCMS: 403.1 [M+H]

[0216] Intermediate A22 5-Iodo-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-D]pyrimidin-4-amine

[0217] Step 1: Synthesis of tert-butyl 3-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylate [ka] The title compound was prepared in a manner similar to that described in Step 1 of Intermediate A15, starting from 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2.00 g, 7.66 mmol) and tert-butyl 3-(tosyloxy)pyrrolidine-1-carboxylate (synthesized as described in PCT International Publication No. WO 2019 / 238067 A1, 2.62 g, 7.66 mmol), and obtained as a yellow solid (2.0 g, 61% yield). LCMS: 431.0 [M+H]

[0218] Step 2: Synthesis of 5-iodo-7-(pyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] The title compound was prepared in a manner similar to that described in Step 2 of Intermediate A15, starting from tert-butyl 3-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylate (1.0 g, 2.324 mmol), and was obtained as an off-white solid (0.70 g, crude), which was used without further purification. LCMS: 331.0 [M+H]

[0219] Step 3: Synthesis of 5-iodo-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] The title compound was prepared in a similar manner as described in Step 2 of Intermediate A18, starting from 5-iodo-7-(pyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (1.0 g, 3.03 mmol), and was obtained as an off-white solid (0.33 g, 32% yield). It was subsequently purified by preparative HPLC (elution: 0.1% HCOOH / water and ACN). LCMS: 345.1 [M+H]

[0220] Intermediate A23 tert-Butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-D]pyrimidin-1-yl)piperidine-1-carboxylate [ka] The title compound was prepared as described in PCT International Publication No. WO 2016 / 112846 A1.

[0221] Intermediate A24 3-Iodo-1-(1-methylpiperidin-4-yl)-1H-pyrazolo[3,4-D]pyrimidin-4-amine [ka] The title compound was prepared as described in J. Med. Chem. 2016, 59(21), 9788-9805.

[0222] Intermediate A25 tert-Butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-D]pyrimidin-1-yl)azetidine-1-carboxylate [ka] The title compound was prepared in a manner similar to that described in Step 1 of Intermediate A15, starting from 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.000 g, 3.83 mmol) and tert-butyl 3-iodoazetidine-1-carboxylate (1.085 g, 3.83 mmol), and obtained as an off-white solid (1.31 g, 82% yield). 1 H NMR(400MHz, DMSO-d6)δ=8.20(s, 1H), 7.84(s, 2H), 5.51-5.76(m, 1H), 4.20-4.35(m, 4H), 1.42(s, 9H); LCMS: 417.0 [M+H]

[0223] Intermediate A26 3-Iodo-1-(1-methylazetidin-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-4-amine [ka] The title compound was prepared as described in PCT International Publication No. WO 2002 / 076986 A1.

[0224] Intermediate A27 3-Iodo-1-(prop-2-yn-1-yl)-1H-pyrazolo[3,4-D]pyrimidin-4-amine [ka] K2CO3 (1.059 g, 7.66 mmol) and 3-bromo-1-propyne (80% solution in toluene, 0.38 mL, 3.83 mmol) were added to a solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.000 g, 3.83 mmol) in DMF (5 mL), and the resulting mixture was stirred at 70 °C for 12 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was added to ice-cold water (50 mL) and extracted with EtOAc (3 × 50 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), eluting with 70% EtOAc / petroleum ether) to give the title compound as a light brown solid (0.846 g, 74% yield). 1 H NMR (400MHz, DMSO-d6)δ=8.25(s, 1H), 6.77(bs, 2H), 5.14(s, 2H), 3.39(s, 1H); LCMS: 300.0 [M+H]

[0225] Intermediate A28 1-(but-3-yn-1-yl)-3-iodo-1H-pyrazolo[3,4-D]pyrimidin-4-amine [ka] The title compound was prepared in a manner similar to that described in Intermediate A27, starting from 4-bromobut-1-yne (0.123 g, 0.9589 mmol) and 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.25 g, 0.958 mmol), and obtained as a light brown solid (0.230 g, 77% yield). 1 H NMR (400MHz, DMSO-d6)δ=8.21(s, 1H), 6.70(bs, 2H), 4.39(t, J=6.8Hz, 2H), 2.72-2.80(m, 3H); LCMS: 314.0 [M+H]

[0226] Intermediate A29 2-(4-amino-5-iodo-7H-pyrrolo[2,3-D]pyrimidin-7-yl)ethan-1-ol [ka] The title compound was prepared as described in PCT International Publication No. WO 2021 / 226547 A2.

[0227] Intermediate B1 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine [ka] A mixture of 1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-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. Pd(PPh3)4 (0.092 g, 0.08 mmol) was then added, and the resulting mixture was stirred at 100 °C for 16 hours. After completion of the reaction (confirmed by TLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), which was then washed with EtOAc (2 × 10 mL). The combined filtrate was concentrated under reduced pressure to give a crude product, which was purified by flash chromatography (silica gel (230-400 mesh), elution: 2% MeOH / DCM) to give the title compound as a yellow solid (0.46 g, 98% yield). 1 H NMR (400MHz, 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]

[0228] Intermediate B2 3-(4-aminophenyl)-1-isopropyl-1H-pyrazolo[3,4-D]pyrimidin-4-amine

[0229] Step 1: Synthesis of 1-isopropyl-3-(4-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine [ka] The title compound was prepared in a manner similar to that described in Intermediate B1, starting from 3-iodo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (A2, 1.887 g, 6.23 mmol) and (4-nitrophenyl)boronic acid (1.560 g, 9.34 mmol), and obtained as a yellow solid (1.242 g, 67% yield). 1 H NMR (400MHz, 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=6.8Hz, 6H); LCMS: 299.1 [M+H]

[0230] Step 2: Synthesis of 3-(4-aminophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine [ka] Iron powder (2.320 g, 41.60 mmol) and NH4Cl (2.220 g, 41.60 mmol) were added to a solution of 1-isopropyl-3-(4-nitrophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.242 g, 4.16 mmol) in ethanol (50 mL) and water (20 mL), and the mixture was stirred at 80 °C for 3 h. After completion of the reaction (as confirmed by TLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), which was then washed with EtOAc (2 × 25 mL). The combined filtrate was concentrated under reduced pressure, and the resulting residue was dissolved in EtOAc (100 mL), washed with brine (25 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the title compound as a pale yellow solid (1.042 g, quantitative yield), which was carried forward without further purification.

[0231] Intermediate B3 3-(4-aminophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-4-amine

[0232] Step 1: Synthesis of 3-(4-nitrophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine [ka] The title compound was prepared in a similar manner as described in Intermediate B1, starting from 3-iodo-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (A3, 0.800 g, 2.522 mmol) and (4-nitrophenyl)boronic acid (0.632 g, 3.78 mmol), and obtained as a yellow solid (0.596 g, 76% yield). 1 H NMR (400MHz, 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]

[0233] Step 2: Synthesis of 3-(4-aminophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine [ka] The title compound was prepared in a manner similar to that described in Step 2 of Intermediate B2, starting from 3-(4-nitrophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.596 g, 1.91 mmol), and was obtained as a pale yellow solid (0.42 g, quantitative yield), which was carried forward without further purification. LCMS: 283.0 [M+H]

[0234] Intermediate B4 1-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-D]pyrimidin-1-yl)-2-methylpropan-2-ol [ka] The title compound was prepared in a similar manner as described in Intermediate B1, starting from 1-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-2-methylpropan-2-ol (A4, 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), and obtained as a pale yellow solid (0.077 g, 66% yield). LCMS: 317.1 [M+H]

[0235] Intermediate B5 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] A mixture of 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A5, 10.5 g, 35.0 mmol), 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (9.12 g, 38.5 mmol), and K2CO3 (9.67 g, 70.0 mmol) in 1,4-dioxane (130 mL), EtOH (60 mL), and water (74.2 mL) was purged with N2 for 10 minutes. PdCl2(dppf) (1.280 g, 1.749 mmol) was then added, and the resulting mixture was stirred at 80 °C for 3 hours. After completion of the reaction (confirmed by TLC), the reaction mixture was filtered through Celite, which was then washed with EtOAc (2 × 100 mL). The combined filtrate was concentrated under reduced pressure to give a crude product, which was purified by GRACE (silica gel (230-400 mesh), elution: 3% MeOH / DCM) to give the title compound as a yellow solid (5.7 g, 57% yield). 1H NMR (400MHz, DMSO-d6)δ=8.13(s, 1H), 7.12(s, 1H), 7.05-7.09(m, 1H), 6.95-6.98(m, 1H), 6.82-6.87(m, 1H), 6.04(bs, 2H), 5.22(bs, 2H), 3.53-3.56(m, 1H), 1.02-1.04(m, 4H); LCMS: 284.1 [M+H]

[0236] Intermediate B6 5-(4-amino-3-fluorophenyl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A6, 0.120 g, 0.438 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.114 g, 0.482 mmol), and obtained as a pale yellow solid (0.09 g, 65% yield). LCMS: 258.1 [M+H]

[0237] Intermediate B7 5-(4-amino-3-fluorophenyl)-7-cyclobutyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from 7-cyclobutyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A7, 0.410 g, 1.305 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.309 g, 1.305 mmol), and obtained as a brown solid (0.18 g, 37% yield). 1H NMR (400MHz, DMSO-d6)δ=8.13(s, 1H), 7.51(s, 1H), 7.09-7.13(m, 1H), 6.98-7.01(m, 1H), 6.84-6.88(m, 1H), 6.21(bs, 2H), 5.14-5.26(m, 3H), 2.50-2.51(m, 2H), 2.34-2.39(m, 2H), 1.85-1.92(m, 2H); LCMS: 298.0 [M+H]

[0238] Intermediate B8 Di-tert-butyl (3-(6-aminopyridin-3-yl)-1-cyclopropyl-1H-pyrazolo[3,4-D]pyrimidin-4-yl)iminodicarbonate [ka] The title compound was prepared in a similar manner to that described in Intermediate B5, starting from 5-iodo-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A9, 0.210 g, 0.612 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.145 g, 0.612 mmol), and obtained as a brown sticky substance (0.048 g, 24% yield). It was subsequently purified by preparative HPLC (elution: 0.1% TFA / HO and ACN). LCMS: 327.3 [M+H]

[0239] Intermediate B9 5-(4-amino-3-fluorophenyl)-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-D]pyrimidin-4-amine [ka] The title compound was prepared in a similar manner to that described for Intermediate B5, starting from 5-iodo-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A10, 0.210 g, 0.388 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.095 g, 0.427 mmol), and obtained as a brown sticky substance (0.08 g, 59% yield). It was subsequently purified by preparative HPLC (elution: 10 mM NHOAc / HO and ACN). LCMS: 341.2 [M+H]

[0240] Intermediate B10 3-(4-aminophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-4-amine

[0241] Step 1: Synthesis of di-tert-butyl (1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)iminodicarbonate [ka] DMAP (0.073 g, 0.598 mmol) and (Boc)2O (1.631 g, 7.47 mmol) were added to a solution of 1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (A1, 0.900 g, 2.99 mmol) in THF (10 mL), and the resulting solution was stirred at 25 °C for 12 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was concentrated under reduced pressure to give the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), elution: 80% EtOAc / petroleum ether) to give the title compound as a light brown solid (0.40 g, 27% yield). LCMS: 501.8 [M+H]

[0242] Step 2: Synthesis of di-tert-butyl (3-(6-aminopyridin-3-yl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)iminodicarbonate [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from di-tert-butyl (1-cyclopropyl-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)iminodicarbonate (0.200 g, 0.399 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (0.097 g, 0.439 mmol), and obtained as a brown sticky material (0.09 g, 48% yield). LCMS: 468.2 [M+H]

[0243] Intermediate B11 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-N-(2,4-dimethoxybenzyl)-1H-pyrazolo[4,3-C]pyridin-4-amine [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from 1-cyclopropyl-N-(2,4-dimethoxybenzyl)-3-iodo-1H-pyrazolo[4,3-c]pyridin-4-amine (A11, 0.370 g, 0.748 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.177 g, 0.748 mmol), and obtained as a brown solid (0.15 g, 48% yield). 1 H NMR (400MHz, DMSO-d6)δ=7.82(d, J=6.0Hz, 1H), 7.07-7.19(m, 3H), 6.83-6.89(m, 2H), 6.51(d, J=2.0Hz, 1H), 6.40-6.43(m, 1H), 5.62(t, J=5.6Hz, 1H), 5.49(bs, 2H), 4.49(d, J=5.6Hz, 2H), 3.65-3.72(m, 7H), 1.08-1.09(m, 4H); LCMS: 434.2 [M+H]

[0244] Intermediate B12 3-(6-aminopyridin-3-yl)-1-cyclopropyl-N-(2,4-dimethoxybenzyl)-1H-pyrazolo[4,3-C]pyridin-4-amine [ka] Starting from 1-cyclopropyl-N-(2,4-dimethoxybenzyl)-3-iodo-1H-pyrazolo[4,3-c]pyridin-4-amine (A11, 0.300 g, 0.666 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (0.147 g, 0.666 mmol), the title compound was prepared in a similar manner as described in Intermediate B5 and obtained as a brown sticky material (0.11 g, 30% yield). LCMS: 416.9 [M+H]

[0245] Intermediate B13 5-(4-amino-3-chlorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A1, 0.250 g, 0.833 mmol) and 2-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.211 g, 0.833 mmol), and obtained as a yellow solid (0.03 g, 12% yield). LCMS: 300.1 [M+H]

[0246] Intermediate B14 2-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-D]pyrimidin-1-yl)ethan-1-ol [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from 2-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethan-1-ol (A12, 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), and obtained as a brown solid (0.39 g, 92% yield). LCMS: 289.1 [M+H]

[0247] Intermediate B15 3-(4-amino-3-fluorophenyl)-1-(2-methoxyethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine [ka] The title compound was prepared in a manner similar to that described in Intermediate B5, starting from 3-iodo-1-(2-methoxyethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (A13, 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), and was obtained as a brown solid (0.42 g), which was used without further purification. LCMS: 302.9 [M+H]

[0248] Intermediate B16 5-(4-amino-3-fluorophenyl)-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] The title compound was prepared in a manner similar to that described in Intermediate B5, starting from 5-iodo-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A14, 0.500 g, 1.572 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.372 g, 1.569 mmol), and obtained as a brown solid (0.236 g, 50% yield), which was used without further purification. LCMS: 302.2 [M+H]

[0249] Intermediate B17 5-(4-amino-3-fluorophenyl)-7-(1-(methylsulfonyl)piperidin-4-yl)-7H-pyrrolo[2,3-D]pyrimidin-4-amine [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from 5-iodo-7-(1-(methylsulfonyl)piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A15, 0.500 g, 0.843 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.220 g, 0.927 mmol), and obtained as a brown solid (0.020 g, 5.9% yield). LCMS: 405.2 [M+H]

[0250] Intermediate B18 5-(4-amino-3-fluorophenyl)-7-(1-(cyclopropylsulfonyl)piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from 7-(1-(cyclopropylsulfonyl)piperidin-4-yl)-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A16, 0.103 g, 0.231 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.060 g, 0.254 mmol), and obtained as a brown solid (0.052 g, 52% yield). LCMS: 431.1 [M+H]

[0251] Intermediate B19 5-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-D]pyrimidin-1-yl)pentan-1-ol [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from 5-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pentan-1-ol (A17, 0.194 g, 0.559 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.070 g, 0.559 mmol), and obtained as a brown sticky material (0.052 g, 38% yield). LCMS: 331.1 [M+H]

[0252] Intermediate B20 tert-Butyl 4-(4-amino-5-(4-amino-3-fluorophenyl)-7H-pyrrolo[2,3-D]pyrimidin-7-yl)butanoate [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from tert-butyl 4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)butanoate (A21, 1.280 g, 3.18 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.754 g, 3.18 mmol), and obtained as a brown solid (0.40 g, 33% yield). LCMS: 386.2 [M+H]

[0253] Intermediate B21 tert-Butyl 3-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-D]pyrimidin-1-yl)piperidine-1-carboxylate [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylate (A23, 1.800 g, 4.05 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.961 g, 4.05 mmol), and obtained as a yellow solid (1.0 g, 58% yield). LCMS: 428.2 [M+H]

[0254] Intermediate B22 tert-Butyl 3-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-D]pyrimidin-1-yl)azetidine-1-carboxylate [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)azetidine-1-carboxylate (A25, 0.650 g, 1.562 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.370 g, 1.562 mmol), and obtained as a yellow solid (0.35 g, 56% yield). 1 H NMR (400MHz, DMSO-d6)δ=8.23(s, 1H), 7.22-7.32(m, 2H), 6.89-6.94(m, 1H), 5.60-5.66(m, 1H), 5.52(s, 2H), 4.30-4.36(m, 4H), 1.42(s, 9H); LCMS: 400.2 [M+H]

[0255] Intermediate B23 3-(4-amino-3-fluorophenyl)-1-(prop-2-yn-1-yl)-1H-pyrazolo[3,4-D]pyrimidin-4-amine [ka] The title compound was prepared in a manner similar to that described in Intermediate B5, starting from 3-iodo-1-(prop-2-yn-1-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (A27, 0.846 g, 2.83 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.671 g, 2.83 mmol), and obtained as a brown solid (0.136 g, 17% yield). 1 H NMR (400MHz, DMSO-d6)δ=8.25(s, 1H), 7.19-7.28(m, 2H), 6.87-6.93(m, 1H), 5.50(s, 2H), 5.15(s, 2H), 3.37(s, 1H); LCMS: 283.0 [M+H]

[0256] Intermediate B24 3-(4-amino-3-fluorophenyl)-1-(but-3-yn-1-yl)-1H-pyrazolo[3,4-D]pyrimidin-4-amine [ka] The title compound was prepared in a similar manner as described in Intermediate B5, starting from 1-(but-3-yn-1-yl)-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (A28, 1.000 g, 3.19 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.757 g, 3.18 mmol), and obtained as a brown solid (0.276 g, 29% yield). 1 H NMR (400MHz, DMSO-d6)δ=8.23(s, 1H), 7.19-7.27(m, 2H), 6.90(t, J=8.4Hz, 1H), 5.49(s, 2H), 4.42(t, J=6.8Hz, 2H), 2.75-2.82(m, 3H); LCMS: 297.2 [M+H]

[0257] Intermediate B25 2-(4-amino-5-(4-amino-3-fluorophenyl)-7H-pyrrolo[2,3-D]pyrimidin-7-yl)ethan-1-ol [ka] The title compound was prepared in a manner similar to that described in Intermediate B5, starting from 2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-ol (A29, 0.50 g, 1.644 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.39 g, 1.644 mmol), and obtained as a brown solid (0.30 g, 63% yield). LCMS: 288.1 [M+H]

[0258] Carbamate intermediate C General procedure for the synthesis of carbamate intermediate C Pyridine (1.2 equiv) and phenyl chloroformate (1.5 equiv) were added to a solution of amine (1.0 equiv) in THF (10 vol) at 0°C. The reaction mixture was warmed to 25°C and stirred for 16 h. Upon completion of the reaction (as determined by TLC), either (i) the precipitated solid was filtered, washed with THF, and dried to give the desired carbamate; or (ii) the reaction mixture was diluted with EtOAc (10 mL), washed with brine (5 mL), and the organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude material, which was purified by flash chromatography (silica gel (230-400 mesh), elution: 10-20% EtOAc / petroleum ether) to give the desired carbamate.

[0259] The following carbamates were prepared using the general method described above. [Table 28] [Table 29] [Table 30] [Table 31] [Table 32]

[0260] All amines used in the synthesis of carbamate intermediate C are commercially available, with the following exceptions: 3-Methyl-4-morpholinoaniline (C4 precursor) was synthesized from 4-(2-methyl-4-nitrophenyl)morpholine as described in J. Med. Chem. 2017, 60(12), 5099-5119. 3-Chloro-4-((4-methylpiperazin-1-yl)methyl)aniline (C9 precursor) was synthesized from (4-amino-2-chlorophenyl)(4-methylpiperazin-1-yl)methanone as described in PCT International Publication No. WO 2020 / 135507 A1.

[0261] [Table 33] [Table 34] [Table 35] [Table 36] [Table 37]

[0262] All intermediate D amines are commercially available except for the following: Intermediate D3 was prepared as described in PCT International Publication No. WO 2018 / 215668 A1. Intermediate D4 was prepared as described in PCT International Publication No. WO 2020 / 206583 A1. Intermediate D5 was prepared as described in PCT International Publication No. WO 2016 / 029776 A1. Intermediate D7 was prepared as described in PCT International Publication No. WO 2017 / 222285 A1. Intermediate D8 was prepared as described in J. Med. Chem. 2012, 55(22), 10033-10046. Intermediate D9 was prepared as described in PCT International Publication No. WO 2020 / 135507 A1. Intermediate D10 was prepared as described in PCT International Publication No. WO 2014 / 012360 A1. Intermediate D11 was prepared as described in PCT International Publication No. WO 2008 / 046802 A1. Intermediate D12 was prepared as described in PCT International Publication No. WO 2019 / 200120 A1. Intermediate D13 was prepared as described in PCT International Publication No. WO 2021 / 067569 A1. Intermediates D14, D15, and D46 were prepared as described in PCT International Publication No. WO 2018 / 102751 A1. Intermediates D18, D24, D35, D47, and D52 were prepared as described in PCT International Publication No. WO 2022 / 008383 A1. Intermediate D20 was prepared as described in J. Med. Chem. 2017, 60(12), 5099-5119. Intermediate D21 was prepared as described in PCT International Publication No. WO 2006 / 066174 A1. Intermediate D25 was prepared as described in PCT International Publication No. WO 2008 / 046802 A1. Intermediate D26 was prepared as described in CN 113717156 A1. Intermediates D30 and D31 were prepared as described in PCT International Publication No. WO 2021 / 226547 A2. Intermediates D32, D38, and D41 were prepared as described in PCT International Publication No. WO 2019 / 200120 A1. Intermediates D33 and D60 were prepared as described in PCT International Publication No. WO 2014 / 072220 A1. Intermediates D34, D49, and D56 were prepared as described in PCT International Publication No. WO 2018 / 215668 A1. Intermediate D39 was prepared as described in J. Med. Chem. 2012, 55(22), 10033-10046. Intermediate D43 was prepared as described in PCT International Publication No. WO 2013 / 170774 A1. Intermediate D48 was prepared as described in PCT International Publication No. WO 2019 / 196812 A1. Intermediates D51 and D58 were prepared as described in PCT International Publication No. WO 2013 / 152198 A1. Intermediate D53 was prepared as described in PCT International Publication No. WO 2017 / 222285 A1. Intermediate D54 was prepared as described in PCT International Publication No. WO 2021 / 164742 A1. Intermediate D59 was prepared as described in PCT International Publication No. WO 2014 / 019338 A1.

[0263] Intermediate D16 was prepared according to the following synthetic scheme. [ka] Intermediate D16 4-((6-methyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-3-(trifluoromethyl)aniline

[0264] Step 1: Synthesis of 2,2,2-trifluoro-N-(4-((6-methyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-3-(trifluoromethyl)phenyl)acetamide CsCO (0.548 g, 1.682 mmol) and 2-methyl-2,6-diazaspiro[3.3]heptane hydrochloride (0.250 g, 1.682 mmol) were added to a solution of N-(4-(bromomethyl)-3-(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (synthesized as described in Org Process Res Dev 2008, 12(6), 1146-1155, 0.589 g, 1.682 mmol) in ACN (5 mL), and the mixture was stirred at 25 °C for 12 h. After completion of the reaction (as confirmed by TLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), and the resulting filtrate was concentrated under reduced pressure to give the title compound (0.7 g, crude), which was used without further purification. LCMS: 382.1 [M+H]

[0265] Step 2: Synthesis of 4-((6-methyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-3-(trifluoromethyl)aniline K2CO3 (0.178 g, 1.285 mmol) was added to a solution of 2,2,2-trifluoro-N-(4-((6-methyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-3-(trifluoromethyl)phenyl)acetamide (0.700 g, 1.836 mmol) in MeOH (2 mL) and water (2 mL), and the resulting solution was stirred at 25 °C for 12 h. After completion of the reaction (confirmed by TLC), the reaction mixture was concentrated under reduced pressure to give the crude material, which was purified by preparative HPLC (elution: 0.1% TFA / water and ACN) to give the title compound as a colorless sticky material (0.075 g, 14% yield). LCMS: 286.2 [M+H]

[0266] Intermediate D17 was prepared according to the following synthetic scheme. [ka] Intermediate D17 4-((7-methyl-4,7-diazaspiro[2.5]octan-4-yl)methyl)-3-(trifluoromethyl)aniline

[0267] Step 1: Synthesis of tert-butyl 4-(4-nitro-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane-7-carboxylate The title compound was prepared in a manner similar to that described in Step 1 of Intermediate D16, starting from 1-(bromomethyl)-4-nitro-2-(trifluoromethyl)benzene (synthesized as described in PCT International Publication No. WO 2014 / 206343 A1, 0.228 g, 0.804 mmol) and tert-butyl 4,7-diazaspiro[2.5]octane-7-carboxylate hydrochloride (0.200 g, 0.804 mmol), and obtained as a white solid (0.091 g, 26% yield). It was subsequently purified by flash chromatography (silica gel (230-400 mesh), elution: 5% EtOAc / petroleum ether). LCMS: 416.2 [M+H]

[0268] Step 2: Synthesis of 4-(4-nitro-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane HCl / 1,4-dioxane (4M, 1.5 mL, 0.335 mmol) was added to a solution of tert-butyl 4-(4-nitro-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane-7-carboxylate (0.139 g, 0.335 mmol) in DCM (6 mL) at 0° C., and the mixture was stirred at 25° C. for 16 hours. After completion of the reaction (confirmed by TLC), the reaction mixture was concentrated under reduced pressure to give the title compound as a light brown solid (0.10 g, 38% yield). LCMS: 316.1 [M+H]

[0269] Step 3: Synthesis of 7-methyl-4-(4-nitro-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane Formaldehyde (37% aqueous solution, 0.070 g, 0.856 mmol), acetic acid (0.031 g, 0.514 mmol), and STAB (0.145 g, 0.685 mmol) were added to a solution of 4-(4-nitro-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane (0.108 g, 0.343 mmol) in THF (8 mL) at 0 °C, and the mixture was stirred at 25 °C for 16 h. After completion of the reaction (confirmed by UPLC), the reaction mixture was quenched with aqueous NaHCO (10%, 5 mL) and extracted with EtOAc (2 × 10 mL). The combined organic extracts were dried over NaSO, filtered, and concentrated under reduced pressure to give the title compound (0.076 g, crude) as a brown sticky material, which was used without further purification. LCMS: 330.1[M+H]

[0270] Step 4: Synthesis of 4-((7-methyl-4,7-diazaspiro[2.5]octan-4-yl)methyl)-3-(trifluoromethyl)aniline Iron powder (0.129 g, 2.308 mmol) and NH4Cl (0.123 g, 2.308 mmol) were added to a suspension of 7-methyl-4-(4-nitro-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane (0.076 g, 0.231 mmol) in ethanol (5 mL) and water (1.25 mL), and the mixture was stirred at 80 °C for 2 h. After completion of the reaction (confirmed by UPLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), which was then washed with DCM (5 mL). The combined filtrate was concentrated under reduced pressure to give the crude material, which was added to DCM (10 mL), washed with brine (2 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (0.055 g, crude) as a light brown sticky material, which was used without further purification. LCMS: 300.1 [M+H]

[0271] Intermediate D19 was prepared according to the following synthetic scheme. [ka] Intermediate D19 4-((4-methylpiperazin-1-yl)methyl)-2-(trifluoromethyl)aniline

[0272] Step 1: Synthesis of (4-nitro-3-(trifluoromethyl)phenyl)methanol NaBH (0.0518 g, 1.369 mmol) was added portionwise to a solution of 4-nitro-3-(trifluoromethyl)benzaldehyde (0.250 g, 1.141 mmol) in THF (8 mL) at 0 °C, and the mixture was stirred at 25 °C for 1 h. After completion of the reaction (confirmed by TLC), the reaction mixture was quenched with saturated NH Cl solution (6 mL) and extracted with EtOAc (2 × 10 mL). The combined organic extracts were dried over Na SO , filtered, and concentrated under reduced pressure to give the title compound (0.259 g, crude), which was used without further purification or characterization.

[0273] Step 2: Synthesis of 4-nitro-3-(trifluoromethyl)benzyl methanesulfonate TEA (0.173 g, 1.709 mmol) and MsCl (0.144 g, 1.254 mmol) were added to a solution of (4-nitro-3-(trifluoromethyl)phenyl)methanol (0.252 g, 1.140 mmol) in DCM (7 mL) at 0 °C, and the mixture was stirred at 25 °C for 1 h. After completion of the reaction (confirmed by TLC), the reaction mixture was diluted with DCM (10 mL) and washed with brine (2 mL). The organic layer was dried over Na SO , filtered, and concentrated under reduced pressure to give the title compound (0.276 g, crude), which was used without further purification or characterization.

[0274] Step 3: Synthesis of 1-methyl-4-(4-nitro-3-(trifluoromethyl)benzyl)piperazine K2CO3 (0.255 g, 1.845 mmol) and 1-methylpiperazine (0.092 g, 0.922 mmol) were added to a solution of 4-nitro-3-(trifluoromethyl)benzylmethanesulfonate (0.276 g, 0.922 mmol) in ACN (6 mL), and the mixture was stirred at 60 °C for 16 h. After completion of the reaction (as confirmed by TLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), which was then washed with ACN (2 × 2 mL). The combined filtrate was concentrated under reduced pressure to give the title compound (0.251 g, 74% yield), which was used without further purification. LCMS: 304.4 [M+H]

[0275] Step 4: Synthesis of 4-((4-methylpiperazin-1-yl)methyl)-2-(trifluoromethyl)aniline Pd / C (10%, 0.025 g) was added to a solution of 1-methyl-4-(4-nitro-3-(trifluoromethyl)benzyl)piperazine (0.251 g, 0.828 mmol) in MeOH (8 mL), and the resulting suspension was stirred under H atmosphere (bladder pressure) at 25° C. for 16 hours. After completion of the reaction (as confirmed by TLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), which was then washed with MeOH (5 mL), and the combined filtrate was concentrated under reduced pressure to give the title compound (0.209 g, 67% yield), which was used without further purification. LCMS: 274.1 [M+H]

[0276] Intermediate D22 was prepared according to the following synthetic scheme. [ka] Intermediate D22 3-Methyl-4-((4-methylpiperazin-1-yl)methyl)aniline

[0277] Step 1: Synthesis of 2-methyl-4-nitrobenzyl methanesulfonate The title compound was prepared in a manner similar to that described in Step 2 of Intermediate D19, starting from (2-methyl-4-nitrophenyl)methanol (synthesized as described in PCT International Publication No. WO 2015 / 117147 A1, 0.240 g, 1.436 mmol) and was used without further purification or characterization (0.250 g, 71% yield).

[0278] Step 2: Synthesis of 1-methyl-4-(2-methyl-4-nitrobenzyl)piperazine The title compound was prepared in a similar manner as described in Step 3 of Intermediate D19, starting from 2-methyl-4-nitrobenzyl methanesulfonate (0.250 g, 1.019 mmol) and 1-methylpiperazine (0.102 g, 1.019 mmol), and was used without further purification (0.254 g, 74% yield). LCMS: 250.3 [M+H]

[0279] Step 3: Synthesis of 3-methyl-4-((4-methylpiperazin-1-yl)methyl)aniline The title compound was prepared in a similar manner as described in Step 4 of Intermediate D19, starting from 1-methyl-4-(2-methyl-4-nitrobenzyl)piperazine (0.240 g, 0.963 mmol) and used without further purification (0.169 g, 80% yield). LCMS: 220.2 [M+H]

[0280] Intermediate D23 was prepared according to the following synthetic scheme. [ka] Intermediate D23 4-((4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline

[0281] Step 1: Synthesis of 1-(2-((tert-butyldiphenylsilyl)oxy)ethyl)-4-(4-nitro-2-(trifluoromethyl)benzyl)piperazine CsCO (0.688 g, 2.112 mmol) and 1-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazine (synthesized as described in PCT International Publication No. WO 2012 / 017251 A1, 0.519 g, 1.408 mmol) were added to a solution of 1-(bromomethyl)-4-nitro-2-(trifluoromethyl)benzene (synthesized as described in PCT International Publication No. WO 2014 / 206343 A1, 0.400 g, 1.408 mmol) in THF (6 mL), and the mixture was stirred at 25° C. for 12 hours. After completion of the reaction (as confirmed by TLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), and the filtrate was concentrated under reduced pressure to provide the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), elution: 10% EtOAc / petroleum ether) to give the title compound as a white solid (0.560 g, 69% yield). LCMS: 572.2 [M+H]

[0282] Step 2: Synthesis of 4-((4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline The title compound was prepared in a manner similar to that described in Step 4 of Intermediate D17, starting from 1-(2-((tert-butyldiphenylsilyl)oxy)ethyl)-4-(4-nitro-2-(trifluoromethyl)benzyl)piperazine (0.560 g, 0.980 mmol) and obtained as a yellow gum (0.530 g, quantitative yield), which was used without further purification. LCMS: 542.3 [M+H]

[0283] Intermediate D27 was prepared according to the following synthetic scheme. [ka] Intermediate D27 5-Amino-2-((4-methylpiperazin-1-yl)methyl)benzonitrile

[0284] Step 1: Synthesis of 2-((4-methylpiperazin-1-yl)methyl)-5-nitrobenzonitrile The title compound was prepared in a similar manner as described in Step 1 of Intermediate D23, starting from 2-(bromomethyl)-5-nitrobenzonitrile (0.100 g, 0.415 mmol) and 1-methylpiperazine (0.042 g, 0.415 mmol), and was obtained as a yellow gum (0.077 g, 71% yield), which was used without further purification. LCMS: 261.1 [M+H]

[0285] Step 2: Synthesis of 5-amino-2-((4-methylpiperazin-1-yl)methyl)benzonitrile The title compound was prepared in a similar manner as described in Step 4 of Intermediate D17, starting from 2-((4-methylpiperazin-1-yl)methyl)-5-nitrobenzonitrile (0.077 g, 0.296 mmol) and was obtained as a yellow gum (0.068 g, 85% yield), which was used without further purification. LCMS: 231.2 [M+H]

[0286] Intermediate D36 2-Fluoro-4-((4-methylpiperazin-1-yl)methyl)aniline [ka] The title compound was prepared in a manner similar to that described in Step 4 of Intermediate D17, starting from 1-(3-fluoro-4-nitrobenzyl)-4-methylpiperazine (synthesized as described in PCT International Publication No. WO 2009 / 002916 A2, 0.125 g, 0.494 mmol), and was obtained as a brown sticky material (0.065 g, 59% yield), which was used without further purification. LCMS: 224.3 [M+H]

[0287] Intermediate D37 2-Fluoro-4-((4-methylpiperazin-1-yl)methyl)aniline [ka] The title compound was prepared in a similar manner as described in Step 4 of Intermediate D17, starting from tert-butyl 4-(4-nitro-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane-7-carboxylate (Step 1 of Intermediate D17, 0.120 g, 0.289 mmol) and obtained as a light brown solid (0.080 g, 72% yield), which was used without further purification. LCMS: 386.2 [M+H]

[0288] Intermediate D40 was prepared according to the following synthetic scheme. [ka] Intermediate D40 4-((2,4-dimethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline

[0289] Step 1: Synthesis of 2,4-dimethyl-1-(4-nitro-2-(trifluoromethyl)benzyl)piperazine The title compound was prepared in a manner similar to that described in Step 1 of Intermediate D23, starting from 1-(bromomethyl)-4-nitro-2-(trifluoromethyl)benzene (0.250 g, 0.880 mmol) and 1,3-dimethylpiperazine (0.101 g, 0.880 mmol), and obtained as a yellow gum (0.270 g, quantitative yield), which was used without further purification. LCMS: 318.1 [M+H]

[0290] Step 2: Synthesis of 4-((2,4-dimethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline The title compound was prepared in a similar manner as described in Step 4 of Intermediate D17, starting from 2,4-dimethyl-1-(4-nitro-2-(trifluoromethyl)benzyl)piperazine (0.287 g, 0.904 mmol) and obtained as a yellow gum (0.074 g, 26% yield) which was used without further purification. LCMS: 288.2 [M+H]

[0291] Intermediate D42 was prepared according to the following synthetic scheme. [ka] Intermediate D42 3-Fluoro-4-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)aniline

[0292] Step 1: Synthesis of 1-fluoro-2-methyl-5-nitro-3-(trifluoromethyl)benzene A mixture of methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (2.57 mL, 20.07 mmol), copper(I) iodide (0.38 g, 2.007 mmol), and 1-fluoro-3-iodo-2-methyl-5-nitrobenzene (synthesized as described in PCT International Publication No. WO 2019 / 044946 A1, 0.940 g, 3.34 mmol) in DMF (8 mL) was purged with N for 15 minutes and stirred at 110 °C for 16 hours. After completion of the reaction (as confirmed by GCMS), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), which was then washed with DCM (20 mL). The combined filtrate was concentrated under reduced pressure to provide the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), elution: petroleum ether) to give the title compound as a colorless liquid (0.461 g, yield 62%). 1 H NMR (400MHz, DMSO-d6)δ=8.46-8.48(m, 1H), 8.28(s, 1H), 2.41(s, 3H); GCMS: 223 [M]

[0293] Step 2: Synthesis of 2-(bromomethyl)-1-fluoro-5-nitro-3-(trifluoromethyl)benzene AIBN (0.068 g, 0.413 mmol) and NBS (0.368 g, 2.066 mmol) were added to a solution of 1-fluoro-2-methyl-5-nitro-3-(trifluoromethyl)benzene (0.461 g, 2.066 mmol) in DCE (25 mL), and the mixture was stirred at 90 °C for 16 h. After completion of the reaction (confirmed by TLC), the reaction mixture was concentrated under reduced pressure to give the crude material. This was purified by flash chromatography (silica gel (230-400 mesh), elution: petroleum ether) to give the title compound as a yellow liquid (0.20 g, 32% yield). 1 H NMR (400MHz, DMSO-d6)δ=8.60-8.63(m, 1H), 8.34(s, 1H), 4.77(s, 2H)

[0294] Step 3: Synthesis of 1-(2-fluoro-4-nitro-6-(trifluoromethyl)benzyl)-4-methylpiperazine The title compound was prepared in a similar manner as described in Step 1 of Intermediate D23, starting from 2-(bromomethyl)-1-fluoro-5-nitro-3-(trifluoromethyl)benzene (0.200 g, 0.662 mmol) and 1-methylpiperazine (0.066 g, 0.662 mmol), and obtained as a yellow gum (0.145 g, 68% yield). 1 H NMR (400MHz, DMSO-d6)δ=8.47-8.51(m, 1H), 8.31(s, 1H), 3.70(s, 2H), 2.42(s, 4H), 2.19-2.34(m, 4H), 2.13(s, 3H); LCMS: 322.1 [M+H]

[0295] Step 4: Synthesis of 3-fluoro-4-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)aniline The title compound was prepared in a similar manner as described in Step 4 of Intermediate D19, starting from 1-(2-fluoro-4-nitro-6-(trifluoromethyl)benzyl)-4-methylpiperazine (0.143 g, 0.445 mmol) and was obtained as a pale brown sticky material (0.074 g, 26% yield), which was used without further purification. LCMS: 292.3 [M+H]

[0296] Intermediate D44 was prepared according to the following synthetic scheme. [ka] Intermediate D44 4-((4-methyl-4,7-diazaspiro[2.5]octan-7-yl)methyl)-3-(trifluoromethyl)aniline

[0297] Step 1: Synthesis of tert-butyl 4-methyl-4,7-diazaspiro[2.5]octane-7-carboxylate The title compound was prepared in a manner similar to that described in Step 3 of Intermediate D17, starting from tert-butyl 4,7-diazaspiro[2.5]octane-7-carboxylate (0.200 g, 0.942 mmol), and was obtained as a colorless sticky material (0.139 g, crude), which was used without further purification. LCMS: 227.1 [M+H]

[0298] Step 2: Synthesis of 4-methyl-4,7-diazaspiro[2.5]octane hydrochloride The title compound was prepared in a similar manner as described in Step 2 of Intermediate A15, starting from tert-butyl 4-methyl-4,7-diazaspiro[2.5]octane-7-carboxylate (0.139 g, 0.614 mmol) and using 1,4-dioxane (3 mL) as solvent, to give an off-white solid (0.080 g, crude), which was used without further purification. LCMS: 127.3 [M+H]

[0299] Step 3: Synthesis of 4-methyl-7-(4-nitro-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane The title compound was prepared in a manner similar to that described in Step 1 of Intermediate D23, starting from 1-(bromomethyl)-4-nitro-2-(trifluoromethyl)benzene (0.150 g, 0.528 mmol) and 4-methyl-4,7-diazaspiro[2.5]octane (0.066 g, 0.528 mmol), and obtained as a yellow gum (0.045 g, 26% yield). LCMS: 330.2 [M+H]

[0300] Step 4: Synthesis of 4-((4-methyl-4,7-diazaspiro[2.5]octan-7-yl)methyl)-3-(trifluoromethyl)aniline The title compound was prepared in a manner similar to that described in Step 2 of Intermediate B2, starting from 4-methyl-7-(4-nitro-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane (0.045 g, 0.137 mmol), and was obtained as a pale yellow solid (0.033 g, 81% yield), which was used without further purification. LCMS: 300.2 [M+H]

[0301] Intermediate D45 was prepared according to the following synthetic scheme. [ka] Intermediate D45 tert-Butyl (1-(4-amino-2-(trifluoromethyl)benzyl)piperidin-4-yl)(methyl)carbamate

[0302] Step 1: Synthesis of tert-butyl methyl (1-(4-nitro-2-(trifluoromethyl)benzyl)piperidin-4-yl)carbamate The title compound was prepared in a manner similar to that described in Step 1 of Intermediate D23, starting from 1-(bromomethyl)-4-nitro-2-(trifluoromethyl)benzene (0.199 g, 0.700 mmol) and tert-butyl methyl(piperidin-4-yl)carbamate (0.150 g, 0.700 mmol), and obtained as a yellow gum (0.284 g, 97% yield). LCMS: 418.2 [M+H]

[0303] Step 2: Synthesis of tert-butyl (1-(4-amino-2-(trifluoromethyl)benzyl)piperidin-4-yl)(methyl)carbamate The title compound was prepared in a manner similar to that described in Step 2 of Intermediate B2, starting from tert-butyl methyl (1-(4-nitro-2-(trifluoromethyl)benzyl)piperidin-4-yl)carbamate (0.285 g, 0.683 mmol), and was obtained as a pale yellow solid (0.260 g, quantitative), which was used without further purification. LCMS: 388.2 [M+H]

[0304] Intermediate D50 was prepared according to the following synthetic scheme. [ka] Intermediate D50 1-(4-(4-amino-2-(trifluoromethyl)phenoxy)piperidin-1-yl)ethan-1-one

[0305] Step 1: Synthesis of 1-(4-(4-nitro-2-(trifluoromethyl)phenoxy)piperidin-1-yl)ethan-1-one TEA (0.425 mL, 3.050 mmol) and acetic anhydride (0.125 mL, 1.321 mmol) were added to a solution of 4-(4-nitro-2-(trifluoromethyl)phenoxy)piperidine (synthesized as described in PCT International Publication No. WO 2018 / 222917 A1, 0.295 g, 1.016 mmol) in DCM (4 mL) at 0 °C, and the mixture was stirred at 25 °C for 16 h. After completion of the reaction (confirmed by TLC), the reaction mixture was quenched with ice water (10 mL) and extracted with DCM (2 × 10 mL). The combined organic extracts were dried over Na SO , filtered, and concentrated under reduced pressure to give the crude material. This was purified by flash column chromatography (silica gel 230-400, elution: 80% EtOAc / petroleum ether) to give the title compound as a yellow sticky material (0.232 g, 70% yield). LCMS: 333.2 [M+H]

[0306] Step 2: Synthesis of 1-(4-(4-amino-2-(trifluoromethyl)phenoxy)piperidin-1-yl)ethan-1-one The title compound was prepared in a similar manner as described in Step 2 of Intermediate B2, starting from 1-(4-(4-nitro-2-(trifluoromethyl)phenoxy)piperidin-1-yl)ethan-1-one (0.230 g, 0.692 mmol), and obtained as a pale brown sticky material (0.131 g, 63% yield). It was subsequently purified by flash column chromatography (silica gel 230-400, elution: 5% MeOH / DCM). LCMS: 303.1 [M+H]

[0307] Intermediates D57a and D57b were prepared according to the following synthetic scheme. [ka] Intermediates D57A and D57B 4-((3-fluoro-1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)aniline

[0308] Step 1: Synthesis of tert-butyl 3-fluoro-4-(4-nitro-2-(trifluoromethyl)phenoxy)piperidine-1-carboxylate Cs2CO3 (0.896 g, 2.750 mmol) was added to a solution of 1-fluoro-4-nitro-2-(trifluoromethyl)benzene (0.250 g, 1.196 mmol) and tert-butyl 3-fluoro-4-hydroxypiperidine-1-carboxylate (0.288 g, 1.315 mmol) in DMF (6 mL), and the mixture was stirred at 100 °C for 16 h. After completion of the reaction (confirmed by TLC), the reaction mixture was added to ice water (60 mL) and extracted with EtOAc (2 × 25 mL). The combined organic extracts were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude material. This was purified by flash column chromatography (silica gel (230-400 mesh), elution: 20% EtOAc / petroleum ether) to give two diastereomers of the title compound as colorless oils (first eluting peak: 0.123 g, 30% yield; second eluting peak: 0.159 g, 39% yield). LCMS of both diastereomers: 309.1 [M+H-Boc]

[0309] Step 2: Synthesis of 3-fluoro-4-(4-nitro-2-(trifluoromethyl)phenoxy)piperidine The two diastereomers of the title compound were prepared in a similar manner as described in Step 2 of Intermediate A15, starting from tert-butyl 3-fluoro-4-(4-nitro-2-(trifluoromethyl)phenoxy)piperidine-1-carboxylate (first diastereomer: 0.123 g, 0.301 mmol; second diastereomer: 0.159 g, 0.389 mmol) and using 1,4-dioxane (3 or 5 mL) as the solvent, to give an off-white solid (first diastereomer: 0.119 g; second diastereomer: 0.125 g; crude hydrochloride salt), which was used without further purification. LCMS for both diastereomers: 309.1 [M+H]

[0310] Step 3: Synthesis of 3-fluoro-1-methyl-4-(4-nitro-2-(trifluoromethyl)phenoxy)piperidine The two diastereomers of the title compound were prepared in a similar manner as described in Step 3 of Intermediate D17, starting from 3-fluoro-4-(4-nitro-2-(trifluoromethyl)phenoxy)piperidine (1st diastereomer: 0.196 g, 0.636 mmol; 2nd diastereomer: 0.093 g, 0.302 mmol) and obtained as a colorless oil (1st diastereomer: 0.189 g; 2nd diastereomer: 0.055 g; crude), which was used without further purification. LCMS: 1st diastereomer 323.2 [M+H]; 2nd diastereomer 323.1 [M+H]

[0311] Step 2: Synthesis of 4-((3-fluoro-1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)aniline The two diastereomers of the title compound were prepared in a similar manner as described in Step 4 of Intermediate D19, starting from 3-fluoro-1-methyl-4-(4-nitro-2-(trifluoromethyl)phenoxy)piperidine (1st diastereomer: 0.220 g, 0.683 mmol; 2nd diastereomer: 0.055 g, 0.683 mmol) and obtained as a light brown sticky material (1st diastereomer: 0.085 g, 42% yield; 2nd diastereomer: 0.045 g, 90% yield), which was used without further purification. LCMS: 1st diastereomer: 293.1 [M+H]; 2nd diastereomer: 293.2 [M+H]

[0312] Intermediate D61 1-(4-amino-2-(trifluoromethyl)benzyl)piperidin-4-ol [ka] The title compound was prepared in a manner similar to that described in Step 4 of Intermediate D19, starting from 1-(4-nitro-2-(trifluoromethyl)benzyl)piperidin-4-ol (0.254 g, 0.835 mmol), and was obtained as a pale brown sticky material (0.173 g, 75% yield), which was used without further purification. LCMS: 275.2 [M+H]

[0313] Intermediate D62 was prepared according to the following synthetic scheme. [ka] Intermediate D62 4-((4-(cyclopropylsulfonyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline Step 1: Synthesis of 1-(cyclopropylsulfonyl)-4-(4-nitro-2-(trifluoromethyl)benzyl)piperazine The title compound was prepared in a manner similar to that described in Step 3 of Intermediate A15, starting from 1-(4-nitro-2-(trifluoromethyl)benzyl)piperazine (synthesized as described in PCT International Publication No. WO 2014 / 072220 A1, 0.408 g, 1.411 mmol) and cyclopropanesulfonyl chloride (0.198 g, 1.411 mmol), and obtained as an off-white solid (0.275 g, 49% yield). LCMS: 394.0 [M+H]

[0314] Step 2: Synthesis of 4-((4-(cyclopropylsulfonyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline The title compound was prepared in a manner similar to that described in Step 4 of Intermediate D19, starting from 1-(cyclopropylsulfonyl)-4-(4-nitro-2-(trifluoromethyl)benzyl)piperazine (0.272 g, 0.691 mmol), and was obtained as a pale brown sticky material (0.225 g, 90% yield), which was used without further purification. LCMS: 364.2 [M+H]

[0315] Intermediate D63 was prepared according to the following synthetic scheme. [ka] Intermediate D63 4-((4-(methylsulfonyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline

[0316] Step 1: Synthesis of 1-(methylsulfonyl)-4-(4-nitro-2-(trifluoromethyl)benzyl)piperazine The title compound was prepared in a similar manner as described in Step 1 of Intermediate D23, starting from 1-(bromomethyl)-4-nitro-2-(trifluoromethyl)benzene (0.250 g, 0.880 mmol) and 1-(methylsulfonyl)piperazine (0.145 g, 0.880 mmol), and obtained as a pale yellow solid (0.060 g, 19% yield). LCMS: 368.1 [M+H]

[0317] Step 2: Synthesis of 4-((4-(methylsulfonyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline The title compound was prepared in a similar manner as described in Step 4 of Intermediate D19, starting from 1-(methylsulfonyl)-4-(4-nitro-2-(trifluoromethyl)benzyl)piperazine (0.050 g, 0.136 mmol), and was obtained as a pale brown sticky material (0.037 g, 81% yield), which was used without further purification. LCMS: 338.2 [M+H]

[0318] Intermediate D64 was prepared according to the following synthetic scheme. [ka] Intermediate D64 4-((4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline

[0319] Step 1: Synthesis of 1-(2-(methylsulfonyl)ethyl)-4-(4-nitro-2-(trifluoromethyl)benzyl)piperazine The title compound was prepared in a similar manner as described in Step 1 of Intermediate D23, starting from 1-(bromomethyl)-4-nitro-2-(trifluoromethyl)benzene (0.250 g, 0.880 mmol) and 1-(2-(methylsulfonyl)ethyl)piperazine dihydrochloride (0.233 g, 0.880 mmol), as a pale yellow sticky material (0.175 g, 50% yield). LCMS: 396.1 [M+H]

[0320] Step 2: Synthesis of 4-((4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline The title compound was prepared in a manner similar to that described in Step 4 of Intermediate D19, starting from 1-(2-(methylsulfonyl)ethyl)-4-(4-nitro-2-(trifluoromethyl)benzyl)piperazine (0.173 g, 0.438 mmol), and was obtained as a pale brown sticky material (0.135 g, 84% yield), which was used without further purification. LCMS: 366.2 [M+H]

[0321] Intermediate D65 was prepared according to the following synthetic scheme. [ka] Intermediate D65 2-(4-(4-amino-2-(trifluoromethyl)benzyl)piperazin-1-yl)ethane-1-sulfonic acid

[0322] Step 1: Synthesis of 2-(4-(4-nitro-2-(trifluoromethyl)benzyl)piperazin-1-yl)ethane-1-sulfonic acid Sodium 2-bromoethane-1-sulfonate (0.292 g, 1.383 mmol) and CsCO (0.676 g, 2.074 mmol) were added to a solution of 1-(4-nitro-2-(trifluoromethyl)benzyl)piperazine (synthesized as described in PCT International Publication No. WO 2014 / 072220 A1, 0.200 g, 0.691 mmol) in water (5 mL) at 0 °C, and the mixture was stirred at 25 °C for 16 h. After completion of the reaction (as confirmed by LCMS), the reaction mixture was directly purified by preparative HPLC (elution: 0.1% HCOOH / water and ACN) to afford the title compound as an off-white solid (0.045 g, 16% yield). LCMS: 398.2 [M+H]

[0323] Step 2: Synthesis of 2-(4-(4-amino-2-(trifluoromethyl)benzyl)piperazin-1-yl)ethane-1-sulfonic acid Zinc dust (0.074 g, 1.132 mmol) and NH4Cl (0.061 g, 1.132 mmol) were added to a solution of 2-(4-(4-nitro-2-(trifluoromethyl)benzyl)piperazin-1-yl)ethane-1-sulfonic acid (0.045 g, 0.113 mmol) in MeOH (6 mL), and the mixture was stirred at 25°C for 12 hours. After completion of the reaction (as confirmed by TLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), and the resulting filtrate was concentrated under reduced pressure to give the crude material. This was purified by preparative HPLC (elution: 10 mM NH4OAc / water and ACN) to give the title compound as a light brown sticky material (0.03 g, 71% yield). LCMS: 368.1 [M+H]

[0324] Preparation of Examples General Urea Formation Procedure for the Synthesis of Examples 1-115 Method A: DMAP (0.05 eq) and DIPEA (1.5 eq) were added to a solution of carbamate intermediate C (1.0 eq) and amine intermediate D (1.0 eq) in THF (10 vol.), and the mixture was stirred at 60° C. for 12 hours in a sealed tube. After completion of the reaction (as confirmed by LCMS), the reaction mixture was concentrated under reduced pressure to give the crude material, which was purified by reverse-phase preparative HPLC to give the title product. Method B: TEA (2.0 equiv.) was added to a mixture of carbamate intermediate C (1.0 equiv.) and amine intermediate D (1.0 equiv.) in THF (5 mL), and the mixture was stirred in a sealed tube at 60° C. for 12 hours. After completion of the reaction (as confirmed by LCMS), the reaction mixture was concentrated under reduced pressure to give the crude material, which was purified by reverse-phase preparative HPLC to give the title product.

[0325] Example 1 1-(4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-D]pyrimidin-3-yl)phenyl)-3-(3-methyl-4-morpholinophenyl)urea [ka] The title compound was obtained as an off-white solid (0.023 g, 13% yield) following the general procedure for urea formation (Method B), starting from 3-(4-aminophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (B2, 0.100 g, 0.372 mmol) and phenyl(3-methyl-4-morpholinophenyl)carbamate (C4, 0.116 g, 0.372 mmol). 1 H NMR (400MHz, DMSO-d6)δ=8.84(bs, 1H), 8.54(bs, 1H), 8.24(bs, 1H), 7.56-7.64(m, 4H), 7.26-7.29(m, 2H), 6.99(d, J=8.4Hz, 1H), 5.04-5.08(m, 1H), 3.73-3.74(m, 4H), 2.79-2.80(m, 4H), 2.26(s, 3H), 1.49(d, J=6.6Hz, 6H); LCMS: 487.2 [M+H]

[0326] Example 2 1-(4-(4-amino-1-(oxetan-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)phenyl)-3-(3-methyl-4-morpholinophenyl)urea [ka] The title compound was obtained as an off-white solid (0.015 g, 10% yield) starting from 3-(4-aminophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (B3, 0.090 g, 0.318 mmol) and phenyl(3-methyl-4-morpholinophenyl)carbamate (C4, 0.099 g, 0.318 mmol) following the general procedure for urea formation (Method B). 1H NMR (400MHz, DMSO-d6)δ=9.04(bs, 1H), 8.72(bs, 1H), 8.25(bs, 1H), 7.62-7.68(m, 4H), 7.26-7.30(m, 2H), 6.99(d, J=8.5Hz, 1H), 5.98-6.05(m, 1H), 5.10(t, J=6.4Hz, 2H), 4.99(t, J=6.8Hz, 2H), 3.72-3.74(m, 4H), 2.78-2.80(m, 4H), 2.26(s, 3H); LCMS: 501.2 [M+H]

[0327] Example 3 1-(4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-D]pyrimidin-3-yl)phenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.038 g, 22% yield) starting from 3-(4-aminophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (B2, 0.080 g, 0.298 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.117 g, 0.298 mmol) following the general procedure for urea formation (Method B). 1 H NMR (400MHz, DMSO-d6)δ=9.33(bs, 1H), 9.26(bs, 1H), 8.24(s, 1H), 8.01(s, 1H), 7.58-7.68(m, 6H), 5.03-5.08(m, 1H), 3.54(s, 2H), 2.34-2.39(m, 8H), 2.17(s, 3H), 1.50(d, J=6.8Hz, 6H); LCMS: 568.2[M+H]

[0328] Example 4 1-(4-(4-amino-1-(oxetan-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)phenyl)-3-(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.020 g, 16% yield) starting from 3-(4-aminophenyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (B3, 0.080 g, 0.22 mmol) and phenyl(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)carbamate (C3, 0.062 g, 0.22 mmol) following the general urea formation procedure (Method B). 1 H NMR (400MHz, DMSO-d6)δ=9.54(bs, 1H), 9.50(bs, 1H), 8.26(s, 1H), 8.22(bs, 1H), 7.90(d, J=6.0Hz, 2H), 7.66-7.73(m, 4H), 7.59(s, 1H), 7.50(s, 1H), 6.00-6.04(m, 1H), 5.11(t, J=6.4Hz, 2H), 5.00(t, J=6.8Hz, 2H), 2.19(s, 3H); LCMS: 549.9[M+H]

[0329] Example 5 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.019 g, 8% yield) following the general urea formation procedure (Method A), starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (B1, 0.100 g, 0.352 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.208 g, 0.528 mmol). 1 H NMR(400MHz, CD3OD)δ=8.41(s, 1H), 8.34-8.38(m, 1H), 7.96-7.96(m, 1H), 7.68-7.91(m, 2H), 7.48-7.55(m, 2H), 3.98-4.01(m, 1H), 3.76(s, 2H), 3.50-3.52(m, 2H), 3.11-3.15(m, 4H), 2.93(s, 3H), 2.48(bs, 2H), 1.36-1.40(m, 2H), 1.20-1.25(m, 2H); LCMS: 584.2 [M+H]

[0330] Example 6 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-(4-methylpiperazine-1-carbonyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.008 g, 6% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C5, 0.090 g, 0.223 mmol) and (4-amino-2-(trifluoromethyl)phenyl)(4-methylpiperazin-1-yl)methanone (D2, 0.064 g, 0.223 mmol). 1H NMR(400MHz, CD3OD)δ=8.33-8.37(m, 1H), 8.30(s, 1H), 8.08(d, J=2.0Hz, 1H), 7.75-7.78(m, 1H), 7.48-7.53(m, 2H), 7.39-7.41(m, 1H), 3.81-3.85(m, 3H), 3.24-3.28(m, 2H), 2.53-2.61(m, 2H), 2.42(s, 2H), 2.37(s, 3H), 1.29-1.33(m, 2H), 1.16-1.21(m, 2H); LCMS: 598.4 [M+H]

[0331] Example 7 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.037 g, 28% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C5, 0.098 g, 0.213 mmol) and 4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D1, 0.067 g, 0.235 mmol). 1 H NMR(400MHz, CD3OD)δ=8.39(s, 1H), 8.33-8.37(m, 1H), 7.95-7.95(m, 1H), 7.68-7.74(m, 2H), 7.48-7.55(m, 2H), 3.95-3.99(m, 1H), 3.77(bs, 2H), 3.35(bs, 2H), 3.28-3.29(m, 2H), 3.14-3.16(m, 4H), 2.52(bs, 2H), 1.35-1.39(m, 5H), 1.20-1.24(m, 2H); LCMS: 598.2 [M+H]

[0332] Example 8 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(3-fluoro-4-(morpholinomethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.018 g, 11% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C5, 0.140 g, 0.305 mmol) and 3-fluoro-4-(morpholinomethyl)aniline (D10, 0.064 g, 0.305 mmol). 1 H NMR(400MHz, CD3OD)δ=8.32-8.36(m, 1H), 8.29(s, 1H), 7.46-7.55(m, 3H), 7.34-7.38(m, 1H), 7.13-7.16(m, 1H), 3.79-3.84(m, 1H), 3.72-3.75(m, 4H), 3.68(s, 2H), 2.61-2.63(m, 4H), 1.29-1.33(m, 2H), 1.17-1.21(m, 2H); LCMS: 521.2 [M+H]

[0333] Example 9 1-(4-(4-amino-1-(2-hydroxy-2-methylpropyl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.010 g, 5% yield) following the general urea formation procedure (Method A) starting from 1-(4-amino-3-(4-amino-3-fluorophenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-2-methylpropan-2-ol (B4, 0.100 g, 0.316 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.187 g, 0.474 mmol). 1 H NMR(400MHz, CD3OD)δ=8.37-8.40(m, 2H), 7.96(d, J=2.0Hz, 1H), 7.70-7.72(m, 2H), 7.56-7.59(m, 2H), 4.48(s, 2H), 3.76(s, 2H), 3.52(bs, 2H), 2.99-3.24(m, 4H), 2.93(s, 3H), 2.50(bs, 2H), 1.32(s, 6H); LCMS: 615.8 [M+H]

[0334] Example 10 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.039 g, 20% yield) following the general urea formation procedure (Method A), starting from 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (B5, 0.100 g, 0.353 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.139 g, 0.353 mmol). 1H NMR(400MHz, CD3OD)δ=8.36(s, 1H), 8.21(t, J=8.0Hz, 1H), 7.96(d, J=2.0Hz, 1H), 7.67-7.73(m, 2H), 7.47(s, 1H), 7.29-7.37(m, 2H), 3.72(s, 2H), 3.67-3.71(m, 1H), 3.49-3.51(m, 2H), 3.04-3.18(m, 4H), 2.93(s, 3H), 2.48(s, 2H), 1.15-1.25(m, 4H); LCMS: 583.3 [M+H]

[0335] Example 11 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.027 g, 13% yield) following the general urea formation procedure (Method A) starting from 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (B5, 0.100 g, 0.353 mmol) and phenyl(4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)phenyl)carbamate (C7, 0.201 g, 0.529 mmol). 1 H NMR(400MHz, CD3OD)δ=8.34(s, 1H), 8.20(t, J=8.4Hz, 1H), 7.95(d, J=2.4Hz, 1H), 7.72-7.75(m, 1H), 7.54(d, J=8.4Hz, 1H), 7.44(s, LCMS: 569.3 [M+H]

[0336] Example 12 1-(4-(4-amino-7-methyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a white solid (0.009 g, 8% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C8, 0.060 g, 0.159 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.048 g, 0.175 mmol). 1 H NMR (400MHz, DMSO-d6)δ=9.41(bs, 1H), 8.70(d, J=2.4Hz, 1H), 8.11-8.23(m, 2H), 8.01(d, J=2.0Hz, 1H), 7.64-7.66(m, 1H), 7.54-7.57(m, 1H), 7.34(s, 1H), 7.23-7.33(m, 2H), 6.16(bs, 2H), 3.74(s, 3H), 3.55(s, 2H), 2.41(bs, 8H), 2.20(s, 3H); LCMS: 556.8 [M+H]

[0337] Example 13 1-(4-(4-amino-7-cyclobutyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a white solid (0.043 g, 15%), starting from 5-(4-amino-3-fluorophenyl)-7-cyclobutyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (B7, 0.135 g, 0.454 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.179 g, 0.454 mmol), following the general urea formation procedure (Method A). 1 H NMR (400MHz, DMSO-d6)δ=9.53(bs, 1H), 8.83(bs, 1H), 8.40(s, 1H), 8.25(t, J=8.4Hz, 1H), 8.02(d, J=1.6Hz, 1H), 7.94(s, 1H), 7.60-7.66(m, 2H), 7.39-7.42(m, 1H), 7.30(d, J=8.4Hz, 1H), 5.24-5.26(m, 1H), 3.88(s, 2H), 3.40-3.43(m, 2H), 2.90-3.03(m, 4H), 2.82(s, 3H), 2.56-2.68(m, 2H), 2.40-2.46(m, 4H), 1.85-1.90(m, 2H); LCMS: 597.2 [M+H]

[0338] Example 14 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-(4-methylpiperazine-1-carbonyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a white solid (0.009 g, 6% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.100 g, 0.248 mmol) and (4-amino-2-(trifluoromethyl)phenyl)(4-methylpiperazin-1-yl)methanone (D2, 0.078 g, 0.273 mmol).1 H NMR(400MHz, CD3OD)δ=8.36(s, 1H), 8.17-8.21(m, 1H), 8.11(d, J=1.6Hz, 1H), 7.79(d, J=8.4Hz, 1H), 7.48-7.50(m, 2H), 7.30-7.37(m, 2H), 3.68-3.72(m, 1H), 3.46(s, 8H), 2.98(s, 3H), 1.18-1.22(m, 4H);LCMS: 597.3 [M+H]

[0339] Example 15 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.009 g, 4% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.168 g, 0.418 mmol) and 4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D1, 0.120 g, 0.418 mmol). 1 H NMR(400MHz, CD3OD)δ=8.36(s, 1H), 8.21(t, J=8.0Hz, 1H), 7.96(d, J=2.0Hz, 1H), 7.66-7.73(m, 2H), 7.48(s, 1H), 7.29-7.37(m, 2H), 3.73(s, 2H), 3.69-3.72(m, 1H), 3.50-3.57(m, 2H), 3.21-3.27(m, 2H), 3.09-3.16(m, 4H), 2.49(bs, 2H), 1.37(t, J=7.2Hz, 3H), 1.18-1.22(m, 4H); LCMS: 597.3 [M+H]

[0340] Example 16 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methyl-1,4-diazepan-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.010 g, 4% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.160 g, 0.397 mmol) and 4-((4-methyl-1,4-diazepan-1-yl)methyl)-3-(trifluoromethyl)aniline (D7, 0.080 g, 0.278 mmol). 1 H NMR(400MHz, CD3OD)δ=8.15-8.21(m, 2H), 7.90(d, J=2.0Hz, 1H), 7.78(d, J=8.4Hz, 1H), 7.69-7.72(m, 1H), 7.27-7.33(m, 2H), 7.21(s, 1H), 3.84(s, 2H), 3.51-3.54(m, 1H), 3.35-3.38(m, 2H), 3.24-3.28(m, 2H), 2.79-2.91(m, 7H), 2.04-2.07(m, 2H), 1.13-1.17(m, 2H), 1.07-1.10(m, 2H); LCMS: 596.9 [M+H]

[0341] Example 17 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-(morpholinomethyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.012 g, 7% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.077 g, 0.191 mmol) and 4-(morpholinomethyl)-3-(trifluoromethyl)aniline (D3, 0.071 g, 0.273 mmol). 1 H NMR (400MHz, DMSO-d6)δ=9.65(bs, 1H), 8.88(bs, 1H), 8.41(s, 1H), 8.20-8.24(m, 1H), 8.09(bs, 1H), 7.70-7.78(m, 2H), 7.70(s, 1H), 7.35-7.39(m, 1H), 7.24-7.28(m, 1H), 4.35(s, 2H), 3.67-3.72(m, 5H), 3.20(bs, 4H), 1.09-1.11(m, 4H); LCMS: 570.3 [M+H]

[0342] Example 18 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((3-(dimethylamino)pyrrolidin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.040 g, 34% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.100 g, 0.193 mmol) and 1-(4-amino-2-(trifluoromethyl)benzyl)-N,N-dimethylpyrrolidin-3-amine (D4, 0.061 g, 0.213 mmol). 1H NMR(400MHz, CD3OD)δ=8.37(s, 1H), 8.21(t, J=8.0Hz, 1H), 8.04(s, 1H), 7.73-7.73(m, 2H), 7.49(s, 1H), 7.30-7.37(m, 2H), 4.15-4.16(m, 2H), 4.02-4.04(m, 1H), 3.71-3.73(m, 1H), 3.40-3.41(m, 1H), 3.24-3.28(m, 2H), 2.92-2.95(m, 7H), 2.45-2.48(m, 1H), 2.18-2.24(m, 1H), 1.18-1.22(m, 4H); LCMS: 597.3 [M+H]

[0343] Example 19 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-(((3AR,6AS)-5-methylhexahydropyrrolo[3,4-C]pyrrol-2(1H)-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.016 g, 13% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-(((3aR,6aS)-5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)-3-(trifluoromethyl)aniline (D8, 0.056 g, 0.186 mmol). 1H NMR (400MHz, CD3OD) δ=8.17-8.20(m, 2H), 7.91(bs, 1H), 7.71-7.73(m, 2H), 7.28-7.33(m, 2H), 7.21(bs, 1H), 3.79(bs, 2H), 3.52-3.53(m, 3H), 3.03(bs, 4H), 2.88(s, 3H), 2.78-2.81(m, 2H), 2.41(bs, 2H), 1.09-1.17(m, 4H); LCMS: 609.3 [M+H]

[0344] Example 20 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-fluoro-4-((4-methylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.005 g, 5% yield) following the general urea formation procedure (Method A) starting from phenyl(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 3-fluoro-4-((4-methylpiperazin-1-yl)methyl)aniline (D9, 0.042 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.20(s, 1H), 8.14-8.18(m, 1H), 7.49-7.53(m, 1H), 7.27-7.36(m, 3H), 7.21(s, 1H), 7.13-7.16(m, 1H), 3.70(s, 2H), 3.50-3.55(m, 1H), 3.15-3.19(m, 4H), 2.80-2.87(m, 7H), 1.06-1.18(m, 4H); LCMS: 533.3 [M+H]

[0345] Example 21 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-fluoro-4-(morpholinomethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.035 g, 17% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.140 g, 0.347 mmol) and 3-fluoro-4-(morpholinomethyl)aniline (D10, 0.073 g, 0.347 mmol). 1 H NMR(400MHz, CD3OD)δ=8.21(s, 1H), 8.17-8.19(m, 1H), 7.58-7.62(m, 1H), 7.39-7.44(m, 1H), 7.27-7.33(m, 2H), 7.23(s, 1H), 7.18-7.20(m, 1H), 3.95(s, 2H), 3.74-3.82(m, 4H), 3.55-3.58(m, 1H), 2.90(bs, 4H), 1.07-1.19(m, 4H); LCMS: 520.2 [M+H]

[0346] Example 22 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((1-methylpiperidin-4-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a white solid (0.040 g, 33% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((1-methylpiperidin-4-yl)methyl)-3-(trifluoromethyl)aniline (D11, 0.051 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.30-8.34(m, 2H), 7.94(d, J=2.4Hz, 1H), 7.64-7.67(m, 1H), 7.50-7.58(m, 2H), 7.40-7.42(m, 1H), 6.90(s, 1H), 3.60-3.62(m, 1H), 3.51-3.54(m, 2H), 2.94-3.00(m, 2H), 2.86(s, 3H), 2.78-2.79(m, 2H), 1.92-1.95(m, 3H), 1.55-1.58(m, 2H), 1.12-1.14(m, 2H), 0.80-0.83(m, 2H); LCMS: 582.3 [M+H]

[0347] Example 23 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-(4-methyl-1H-imidazol-1-yl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a white solid (0.010 g, 5% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.160 g, 0.397 mmol) and 4-(4-methyl-1H-imidazol-1-yl)-3-(trifluoromethyl)aniline (D5, 0.096 g, 0.397 mmol). 1H NMR(400MHz, CD3OD)δ=8.20-8.22(m, 2H), 8.15-8.15(m, 1H), 7.79-7.82(m, 1H), 7.66(s, 1H), 7.45(d, J=8.4Hz, 1H), 7.29-7.34(m, LCMS: 551.1 [M+H]

[0348] Example 24 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-chloro-4-((4-methylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as an off-white sticky material (0.010 g, 8% yield) following the general urea formation procedure (Method A) starting from 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (B5, 0.060 g, 0.212 mmol) and phenyl(3-chloro-4-((4-methylpiperazin-1-yl)methyl)phenyl)carbamate (C9, 0.084 g, 0.233 mmol). 1 H NMR(400MHz, CD3OD)δ=8.37(s, 1H), 8.22(t, J=8.4Hz, 1H), 7.74(d, J=2.4Hz, 1H), 7.49(s, 1H), 7.43(d, J=8.4Hz, 1H), 7.33-7.44(m, LCMS: 549.3 [M+H]

[0349] Example 25 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a light brown solid (0.100 g, 44% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.150 g, 0.372 mmol) and 4-((1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)aniline (D12, 0.102 g, 0.372 mmol). 1 H NMR(400MHz, CD3OD)δ=8.36(s, 1H), 8.19(t, J=8.4Hz, 1H), 7.88(d, J=2.4Hz, 1H), 7.63-7.66(m, 1H), 7.48(s, 1H), 7.24-7.36(m, 3H), 4.96(bs, 1H), 3.69-3.72(m, 2H), 3.49-3.51(m, 1H), 3.23-3.30(m, 2H), 2.93(bs, 3H), 1.90-2.48(m, 4H), 1.18-1.23(m, 4H); LCMS: 584.3 [M+H]

[0350] Example 26 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(2-fluoro-4-(morpholinomethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.009 g, 9.7% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 2-fluoro-4-(morpholinomethyl)aniline (D10, 0.039 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.20-8.24(m, 2H), 8.07-8.11(m, 1H), 7.26-7.32(m, 2H), 7.15-7.22(m, 3H), 3.71-3.73(m, 4H), 3.50-3.52(m, 3H), 2.48-2.50(m, 4H), 1.14-1.16(m, 2H), 1.08-1.09(m, 2H); LCMS: 520.2 [M+H]

[0351] Example 27 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((3-methyl-3,8-diazabicyclo[3.2.1]octan-8-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.008 g, 10% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.054 g, 0.134 mmol) and 4-((3-methyl-3,8-diazabicyclo[3.2.1]octan-8-yl)methyl)-3-(trifluoromethyl)aniline (D14, 0.040 g, 0.134 mmol). 1H NMR(400MHz, CD3OD)δ=8.35(s, 1H), 8.22(t, J=8.0Hz, 1H), 7.95(s, 1H), 7.86(d, J=8.4Hz, 1H), 7.71(d, J=8.0Hz, 1H), 7.46(s, 1H), 7.30-7.37(m, 2H), 3.66-3.81(m, 3H), 3.49-3.51(m, 2H), 3.15(s, 2H), 2.86(s, 3H), 2.32-2.34(m, 2H), 1.94-1.96(m, 2H), 1.17-1.35(m, 6H); LCMS: 609.5 [M+H]

[0352] Example 28 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-(1-(4-methylpiperazin-1-yl)ethyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.020 g, 16% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.085 g, 0.211 mmol) and 4-(1-(4-methylpiperazin-1-yl)ethyl)-3-(trifluoromethyl)aniline (D15, 0.061 g, 0.211 mmol). 1H NMR(400MHz, CD3OD)δ=8.25(t, J=8.4Hz, 1H), 8.16(s, 1H), 7.91(bs, 1H), 7.80(d, J=8.8Hz, 1H), 7.69(d, J=8.8Hz, 1H), 7.47-7.52(m, 2H), 6.64(s, 1H), 3.69-3.70(m, 1H), 3.47-3.51(m, 1H), 2.80-2.88(m, 4H), 2.49-2.60(m, 7H), 1.34-1.35(m, 3H), 1.05-1.10(m, 2H), 0.66-0.70(m, 2H); LCMS: 597.3 [M+H]

[0353] Example 29 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((6-methyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.002 g, 1.3% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.106 g, 0.263 mmol) and 4-((6-methyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-3-(trifluoromethyl)aniline (D16, 0.075 g, 0.263 mmol). 1H NMR(400MHz, CD3OD)δ=8.31-8.35(m, 2H), 8.12(d, J=2.0Hz, 1H), 7.80-7.83(m, 1H), 7.53-7.64(m, 3H), 6.91(s, 1H), 4.50(bs, 2H), 4.43(bs, 4H), 3.61-3.64(m, 3H), 2.93(s, 3H), 2.45-2.50(m, 2H), 1.05-1.10(m, 2H), 0.66-0.70(m, 2H); LCMS: 595.4 [M+H]

[0354] Example 30 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((7-methyl-4,7-diazaspiro[2.5]octan-4-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.006 g, 5% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((7-methyl-4,7-diazaspiro[2.5]octan-4-yl)methyl)-3-(trifluoromethyl)aniline (D17, 0.056 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.34(s, 1H), 8.21(t, J=8.4Hz, 1H), 7.95(bs, 1H), 7.60-7.66(m, 2H), 7.44(s, 1H), 7.29-7.38(m, 2H), 4.19-4.23(m, 1H), 3.93-3.96(m, 1H), 3.65-3.69(m, 3H), 3.15-3.22(m, 2H), 2.80-2.95(m, 5H), 1.16-1.21(m, 4H), 0.77-0.93(m, 4H); LCMS: 609.6 [M+H]

[0355] Example 31 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((5-methyl-2,5-diazabicyclo[2.2.2]octan-2-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.007 g, 5.8% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.080 g, 0.198 mmol) and 4-((5-methyl-2,5-diazabicyclo[2.2.2]octan-2-yl)methyl)-3-(trifluoromethyl)aniline (D18, 0.059 g, 0.198 mmol). 1 H NMR(400MHz, CD3OD)δ=8.15-8.21(m, 2H), 7.91(bs, 1H), 7.73-7.76(m, 1H), 7.66-7.69(m, 1H), 7.27-7.33(m, 2H), 7.21(s, 1H), 3.89(s, 2H), 3.50-3.54(m, 1H), 3.27-3.30(m, 2H), 3.12-3.15(m, 2H), 2.77-2.81(m, 5H), 1.93-2.12(m, 3H), 1.69-1.70(m, 1H), 1.08-1.17(m, 4H); LCMS: 609.3 [M+H]

[0356] Example 32 1-(5-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)pyridin-2-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka]

[0357] Step 1: Synthesis of 1-(5-bromopyridin-2-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was obtained as an off-white solid (0.035 g, 28% yield) following the general urea formation procedure (Method A) starting from 5-bromopyridin-2-amine (0.044 g, 0.254 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.100 g, 0.254 mmol). It was subsequently purified by preparative HPLC (elution: 0.1% TFA / water and ACN). LCMS: 472.1 [M+H]

[0358] Step 2: Synthesis of tert-butyl (tert-butoxycarbonyl)(7-cyclopropyl-5-(6-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate The title compound was prepared in a manner similar to that described for Intermediate B5, starting from 1-(5-bromopyridin-2-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (0.033 g, 0.070 mmol) and tert-butyl (tert-butoxycarbonyl) (7-cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (prepared as described in IN 2016 / 11024887 A, 0.035 g, 0.070 mmol), and obtained as a brown sticky substance (0.025 g, 46% yield). It was subsequently purified by preparative HPLC (elution: 10 mM NHOAc / HO and ACN). LCMS: 766.8 [M+H]

[0359] Step 3: Synthesis of 1-(5-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pyridin-2-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea TFA (0.125 mL) was added to a solution of tert-butyl (tert-butoxycarbonyl)(7-cyclopropyl-5-(6-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (0.025 g, 0.033 mmol) in DCM (2 mL) at 0° C., and the resulting mixture was stirred at 25° C. for 12 hours. After completion of the reaction (as confirmed by LCMS), the reaction mixture was concentrated under reduced pressure to give the crude material, which was purified by preparative HPLC (elution: 10 mM NH4OAc / HO and ACN) to give the title product as an off-white solid (0.003 g, 16% yield). 1 H NMR(400MHz, CD3OD)δ=8.47(bs, 1H), 8.22(bs, 1H), 8.03(bs, 1H), 7.87-7.89(m, 1H), 7.74(bs, 2H), 7.35-7.37(m, 1H), 7.27(s, 1H), 3.70(bs, 4H), 3.59(bs, 3H), 2.78(bs, 2H), 2.62(bs, 2H), 2.50(s, 3H), 1.10-1.17(m, 4H); LCMS: 566.3 [M+H]

[0360] Example 33 1-(4-(4-amino-7-(2-hydroxy-2-methylpropyl)-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka]

[0361] Step 1: Synthesis of tert-butyl (tert-butoxycarbonyl) (7-(2-hydroxy-2-methylpropyl)-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate The title compound was prepared in a manner similar to that described in Step 1 of Intermediate B10, starting from 1-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol (A8, 0.400 g, 1.204 mmol), and was obtained as a light brown solid (0.400 g, 61% yield). LCMS: 533.1 [M+H] Step 2: Synthesis of tert-butyl (tert-butoxycarbonyl)(7-(2-hydroxy-2-methylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate TEA (0.348 mL, 2.479 mmol) and XPhos (0.030 g, 0.062 mmol) were added to a solution of tert-butyl (tert-butoxycarbonyl)(7-(2-hydroxy-2-methylpropyl)-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (0.330 g, 0.620 mmol) and 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.357 mL, 2.479 mmol) in 1,4-dioxane (10 mL), and the mixture was purged with N for 10 min. Pd(dba) (0.057 g, 0.062 mmol) was then added, and the mixture was stirred at 100 °C for 6 h. After completion of the reaction (as confirmed by TLC), the reaction mixture was filtered through Celite® (i.e., diatomaceous earth), which was washed with DCM (2×20 mL). The combined filtrate was concentrated under reduced pressure to give the crude material, which was purified by flash chromatography (silica gel (230-400 mesh), elution: 75% EtOAc / petroleum ether) to give the title compound as a pale yellow oil (0.100 g, 30% yield). LCMS: 533.2 [M+H]

[0362] Step 3: Synthesis of tert-butyl (tert-butoxycarbonyl)(5-(3-fluoro-4-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)phenyl)-7-(2-hydroxy-2-methylpropyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate The title compound was prepared in a similar manner to that described for Intermediate B5, starting from 1-(5-bromopyridin-2-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (0.092 g, 0.188 mmol) and tert-butyl (tert-butoxycarbonyl)(7-(2-hydroxy-2-methylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (0.100 g, 0.188 mmol), and obtained as a brown sticky substance (0.030 g, 20% yield). It was subsequently purified by preparative HPLC (elution: 0.1% TFA / HO and ACN). LCMS: 815.3 [M+H]

[0363] Step 4: Synthesis of 1-(4-(4-amino-7-(2-hydroxy-2-methylpropyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was prepared in a manner similar to that described in Step 3 of Example 32, starting from tert-butyl (tert-butoxycarbonyl)(5-(3-fluoro-4-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)phenyl)-7-(2-hydroxy-2-methylpropyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (0.030 g, 0.037 mmol), and obtained as an off-white solid (0.007 g, 31% yield). It was subsequently purified by preparative HPLC (elution: 0.1% TFA / HO and ACN). 1H NMR (400MHz, DMSO-d6)δ=9.50(bs, 1H), 8.79(bs, 1H), 8.37(s, 1H), 8.25(t, J=8.4Hz, 1H), 8.02(d, J=2.0Hz, 1H), 7.59-7.66(m, 2H), 7.50(s, 1H), 7.28-7.38(m, 1H), 7.25-7.26(m, 1H), 4.85(bs, 1H), 4.17(s, 2H), 3.65(s, 2H), 3.35(bs, 2H), 3.02-3.04(m, 2H), 2.77(bs, 2H), 2.69(s, 3H), 2.33-2.35(m, 2H), 1.11(s, 6H); LCMS: 615.3 [M+H]

[0364] Example 34 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-2-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.004 g, 3.7% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-2-(trifluoromethyl)aniline (D19, 0.051 g, 0.186 mmol). 1H NMR(400MHz, CD3OD)δ=7.49-7.55(m, 2H), 7.12(d, J=8.0Hz, 1H), 6.91(d, J=1.6Hz, 1H), 6.84(d, J=8.0Hz, 1H), 6.69-6.78(m, 2H), 6.07(s, 1H), 2.91(s, 2H), 2.79-2.82(m, 1H), 2.48-2.51(m, 4H), 2.24(bs, 2H), 2.10(s, 3H), 1.72(bs, 2H), 0.31-0.33(m, 2H), 0.01-0.03(m, 2H); LCMS: 583.3 [M+H]

[0365] Example 35 1-(4-(4-amino-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.003 g, 3.6% yield) starting from 5-(4-amino-3-fluorophenyl)-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (B8, 0.048 g, 0.147 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.057 g, 0.145 mmol) following the general urea formation procedure (Method A). 1 H NMR(400MHz, CD3OD)δ=8.38(s, 1H), 8.21-8.27(m, 1H), 7.97(s, 1H), 7.66-7.73(m, 2H), 7.31-7.38(m, 2H), 5.59-5.62(m, 1H), 4.08(bs, 2H), 3.50-3.77(m, 7H), 3.06-3.13(m, 7H), 2.92-2.93(m, 5H), 2.54(bs, 2H); LCMS: 626.3 [M+H]

[0366] Example 36 1-(4-(4-amino-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a white solid (0.015 g, 10% yield) starting from 5-(4-amino-3-fluorophenyl)-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (B9, 0.080 g, 0.235 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.092 g, 0.235 mmol) following the general urea formation procedure (Method A). 1 H NMR(400MHz, CD3OD)δ=8.38(s, 1H), 8.23(t, J=8.0Hz, 1H), 7.98(d, J=2.0Hz, 1H), 7.66-7.73(m, 2H), 7.63(s, 1H), 7.32-7.40(m, 2H), 5.08-5.14(m, 1H), 3.74-3.77(m, 4H), 3.37-3.38(m, 10H), 2.99(s, 3H), 2.93(s, 3H), 2.53-2.62(m, 2H), 2.38-2.41(m, 2H); LCMS: 640.4 [M+H]

[0367] Example 37 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(2-fluoro-4-(morpholinomethyl)phenyl)urea [ka] The title compound was obtained as a white solid (0.009 g, 7.9% yield) starting from phenyl (4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C5, 0.090 g, 0.223 mmol) and 2-fluoro-4-(morpholinomethyl)aniline (D10, 0.047 g, 0.223 mmol) following the general urea formation procedure (Method A). 1 H NMR(400MHz, CD3OD)δ=8.36-8.42(m, 3H), 7.47-7.55(m, 2H), 7.33-7.42(m, 2H), 4.36(s, 2H), 3.95-4.06(m, 2H), 3.73-3.78(m, 3H), 3.34-3.37(m, 2H), 3.22-3.33(m, 2H), 1.35-1.41(m, 2H), 1.21-1.25(m, 2H); LCMS: 521.2 [M+H]

[0368] Example 38 1-(5-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-D]pyrimidin-3-yl)pyridin-2-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka]

[0369] Step 1: Synthesis of tert-butyl (tert-butoxycarbonyl)(1-cyclopropyl-3-(6-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)carbamate The title compound was obtained as an off-white solid (0.028 g, 8.5% yield) following the general urea formation procedure (Method A) starting from di-tert-butyl (3-(6-aminopyridin-3-yl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)iminodicarbonate (B10, 0.200 g, 0.428 mmol) and phenyl (4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.185 g, 0.471 mmol). LCMS: 767.3 [M+H]

[0370] Step 2: Synthesis of 1-(5-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)pyridin-2-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was prepared in a manner similar to that described in Step 3 of Example 32, starting from tert-butyl (tert-butoxycarbonyl)(1-cyclopropyl-3-(6-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)carbamate (0.028 g, 0.037 mmol) as an off-white solid (0.019 g, 92% yield). 1 H NMR(400MHz, CD3OD)δ=8.63(d, J=2.0Hz, 1H), 8.40(s, 1H), 8.04-8.08(m, 2H), 7.79-7.81(m, 1H), 7.74(d, J=8.4Hz, 1H), 7.49(d, J=8.8Hz, 1H), 3.96-3.99(m, 1H), 3.78(s, 2H), 3.50-3.51(m, 2H), 3.15-3.32(m, 4H), 2.93(s, 3H), 2.51(bs, 2H), 1.35-1.38(m, 2H), 1.21-1.25(m, 2H); LCMS: 566.9 [M+H]

[0371] Example 39 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[4,3-C]pyridin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka]

[0372] Step 1: Synthesis of 1-(4-(1-cyclopropyl-4-((2,4-dimethoxybenzyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was obtained as an off-white solid (0.030 g, 12% yield) following the general urea formation procedure (Method A) starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-N-(2,4-dimethoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (B11, 0.150 g, 0.346 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.136 g, 0.346 mmol). LCMS: 733.3 [M+H]

[0373] Step 2: Synthesis of 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea Triethylsilane (0.012 g, 0.102 mmol) and TFA (0.004 g, 0.034 mmol) were added to a solution of 1-(4-(1-cyclopropyl-4-((2,4-dimethoxybenzyl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (0.025 g, 0.034 mmol) in DCM (2 mL) at 0° C., and the mixture was stirred at 25° C. for 12 hours. After completion of the reaction (confirmed by LCMS), the reaction mixture was concentrated under reduced pressure to give the crude material, which was purified by preparative HPLC (elution: 0.1% TFA / HO and ACN) to give the title product as an off-white solid (0.010 g, 50% yield). 1 H NMR (400MHz, DMSO-d6)δ=9.64(bs, 1H), 8.99(bs, 1H), 8.36(t, J=8.4Hz, 1H), 7.95-8.03(m, 3H), 7.84(d, J=7.2Hz, 1H), 7.61-7.67(m, 2H), 7.52-7.55(m, 1H), 7.44(d, J=8.4Hz, 1H), 7.32(d, J=7.2Hz, 1H), 3.93-3.99(m, 1H), 3.65(bs, 2H), 3.38-3.40(m, 2H), 2.90-3.03(m, 4H), 2.81(s, 3H), 2.34-2.39(m, 2H), 1.19-1.20(m, 4H); LCMS: 583.3 [M+H]

[0374] Example 40 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-methyl-4-morpholinophenyl)urea [ka] The title compound was obtained as an off-white solid (0.090 g, 61% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.150 g, 0.293 mmol) and 4-((7-methyl-4,7-diazaspiro[2.5]octan-4-yl)methyl)-3-(trifluoromethyl)aniline (D20, 0.056 g, 0.293 mmol). 1 H NMR (400MHz, DMSO-d6)δ=8.99(bs, 1H), 8.72(bs, 1H), 8.42(s, 1H), 8.35(t, J=8.4Hz, 1H), 7.26-7.29(m, 1H), 7.14-7.18(m, 2H), 7.01(d, J=8.8Hz, 1H), 6.96(s, 1H), 6.76(t, J=8.0Hz, 1H), 3.68-3.75(m, 5H), 2.79-2.81(m, 4H), 2.27(s, 3H), 1.03-1.06(m, 2H), 0.67-0.71(m, 2H); LCMS: 502.3 [M+H]

[0375] Example 41 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-chlorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka]

[0376] Step 1: Synthesis of 1-(4-bromo-2-chlorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was prepared according to the general urea formation procedure (Method A) starting from 4-bromo-2-chloroaniline (0.100 g, 0.484 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.191 g, 0.484 mmol) and obtained as an off-white solid (0.030 g, 12% yield) after purification by preparative HPLC (elution: 10 mM NHOAc / water and ACN). LCMS: 505.1 [M+H]

[0377] Step 2: Synthesis of di-tert-butyl(5-(3-chloro-4-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)phenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)iminodicarbonate The title compound was prepared in a manner similar to that described for Intermediate B5, starting from 1-(4-bromo-2-chlorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (0.030 g, 0.060 mmol) and tert-butyl (tert-butoxycarbonyl) (7-cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (prepared as described in IN 2016 / 11024887 A, 0.030 g, 0.060 mmol), and obtained as a brown sticky substance (0.020 g, 42% yield). It was subsequently purified by preparative HPLC (elution: 0.1% TFA / HO and ACN). LCMS: 799.3 [M+H]

[0378] Step 3: Synthesis of 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-chlorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was prepared in a manner similar to that described in Step 3 of Example 32, starting from tert-butyl (tert-butoxycarbonyl)(5-(3-chloro-4-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)phenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (0.020 g, 0.025 mmol) and obtained as an off-white solid (0.42 mg, 2.8% yield). LCMS: 599.2 [M+H]

[0379] Example 42 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-methoxy-4-((4-methylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as a light brown solid (0.037 g, 26% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.106 g, 0.262 mmol) and 3-methoxy-4-((4-methylpiperazin-1-yl)methyl)aniline (D21, 0.062 g, 0.262 mmol). 1 H NMR(400MHz, CD3OD)δ=8.27(t, J=8.0Hz, 1H), 8.16(s, 1H), 7.47-7.53(m, 2H), 7.41(d, J=2.0Hz, 1H), 7.25(d, J=8.4Hz, 1H), 6.92-6.94(m, 1H), 6.64(s, 1H), 3.89(s, 3H), 3.71(s, 2H), 3.49-3.51(m, 1H), 2.74(bs, 8H), 2.45(s, 3H), 1.07-1.09(m, 2H), 0.69-0.70(m, 2H); LCMS: 545.3 [M+H]

[0380] Example 43 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-methyl-4-((4-methylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as a pale yellow solid (0.020 g, 15% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.100 g, 0.248 mmol) and 3-methyl-4-((4-methylpiperazin-1-yl)methyl)aniline (D22, 0.054 g, 0.248 mmol). 1 H NMR (400MHz, DMSO-d6)δ=9.16(bs, 1H), 8.80(bs, 1H), 8.41(s, 1H), 8.34(t, J=8.4Hz, 1H), 7.63-7.67(m, 1H), 7.46-7.48(m, 1H), 7.29-7.31(m, 2H), 7.10-7.23(m, 1H), 6.95(s, 1H), 3.56-3.72(m, 3H), 2.94-2.98(m, 8H), 2.78(s, 3H), 2.34(s, 3H), 1.02-1.04(m, 2H), 0.66-0.67(m, 2H); LCMS: 529.5 [M+H]

[0381] Example 44 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka]

[0382] Step 1: Synthesis of 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was obtained as an off-white solid (0.050 g, 31% yield) following the general urea formation procedure (Method A) starting from phenyl(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and phenyl(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (D23, 0.101 g, 0.186 mmol). LCMS: 852.3 [M+H]

[0383] Step 2: Synthesis of 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea TBAF (1 M, THF solution, 0.053 mL, 0.037 mmol) was added to a solution of 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (0.045 g, 0.053 mmol) in THF (5 mL) at 0° C., and the solution was stirred at 25° C. for 1 hour. After completion of the reaction (confirmed by LCMS), the reaction mixture was concentrated under reduced pressure to give the crude material. This was purified by preparative HPLC (elution: 0.1% TFA / water and ACN) to give the title compound as a white solid (0.010 g, 32% yield). 1H NMR(400MHz, CD3OD)δ=8.37(s, 1H), 8.22(t, J=8.4Hz, 1H), 7.96(d, J=2.0Hz, 1H), 7.69-7.74(m, 2H), 7.50(s, 1H), 7.29-7.37(m, 2H), 3.89-3.91(m, 2H), 3.70-3.73(m, 3H), 3.58(bs, 2H), 3.10-3.29(m, 4H), 3.03(bs, 2H), 2.57-2.60(m, 2H), 1.18-1.32(m, 4H); LCMS: 613.2 [M+H]

[0384] Example 45 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(2-methoxyethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.043 g, 37% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-2-(trifluoromethyl)aniline (D24, 0.059 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.37(s, 1H), 8.22(t, J=8.4Hz, 1H), 7.96(d, J=2.0Hz, 1H), 7.67-7.73(m, 2H), 7.50(s, 1H), 7.29-7.38(m, 2H), 3.70-3.77(m, 5H), 3.43(s, 3H), 3.37-3.38(m, 2H), 2.61-3.03(m, 8H), 1.18-1.23(m, 4H); LCMS: 627.3 [M+H]

[0385] Example 46 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[4,3-C]pyridin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 39, starting from 3-(6-aminopyridin-3-yl)-1-cyclopropyl-N-(2,4-dimethoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (B12, 0.110 g, 0.264 mmol) and phenyl(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C2, 0.108 g, 0.264 mmol) in Step 1, and obtained as an off-white solid (0.002 g, 1.3% yield over two steps). 1 H NMR(400MHz, CD3OD)δ=8.64(d, J=1.6Hz, 1H), 8.06-8.08(m, 2H), 7.80(d, J=6.4Hz, 1H), 7.74-7.75(m, 2H), 7.45(d, J=8.0Hz, 1H), 7.03(d, J=6.4Hz, 1H), 3.68-3.71(m, 3H), 2.51-2.57(m, 8H), 2.02-2.08(m, 2H), 1.22-1.31(m, 4H), 1.13-1.17(m, 3H); LCMS: 580.2 [M+H]

[0386] Example 47 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((1-methylpiperidin-4-ylidene)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a white solid (0.006 g, 11% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.040 g, 0.099 mmol) and 4-((1-methylpiperidin-4-ylidene)methyl)-3-(trifluoromethyl)aniline (D25, 0.027 g, 0.099 mmol). 1 H NMR(400MHz, CD3OD)δ=8.28(t, J=8.0Hz, 1H), 8.16(s, 1H), 7.95(d, J=2.4Hz, 1H), 7.65-7.67(m, 1H), 7.48-7.55(m, 2H), 7.27(d, J=8.4Hz, 1H), 6.64(s, 1H), 6.53(s, 1H), 3.48-3.52(m, 1H), 2.86-2.88(m, 2H), 2.72-2.75(m, 2H), 2.55-2.58(m, 5H), 2.41-2.44(m, 2H), 1.06-1.11(m, 2H), 0.67-0.71(m, 2H); LCMS: 580.3 [M+H]

[0387] Example 48 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(2-methoxy-4-((4-methylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as a light brown solid (0.024 g, 23% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 2-methoxy-4-((4-methylpiperazin-1-yl)methyl)aniline (D26, 0.044 g, 0.186 mmol). 1H NMR(400MHz, CD3OD)δ=8.36(t, J=8.4Hz, 1H), 8.31(s, 1H), 8.15(d, J=8.4Hz, 1H), 7.49-7.57(m, 2H), 7.08(d, J=1.6Hz, 1H), 6.97-6.99(m, 1H), 6.90(s, 1H), 3.97(s, 3H), 3.83(s, 2H), 3.60-3.64(m, 1H), 2.92-2.95(m, 8H), 2.87(s, 3H), 1.11-1.14(m, 2H), 0.82-0.84(m, 2H); LCMS: 545.3 [M+H]

[0388] Example 49 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-cyano-4-((4-methylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as a light brown solid (0.004 g, 4% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 5-amino-2-((4-methylpiperazin-1-yl)methyl)benzonitrile (D27, 0.043 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.31-8.37(m, 2H), 8.05(d, J=2.4Hz, 1H), 7.65-7.68(m, 1H), 7.51-7.59(m, 3H), 6.90(s, 1H), 3.78(s, 2H), 3.61-3.64(m, 1H), 3.49-3.51(m, 2H), 3.13(s, 4H), 2.92(s, 3H), 2.54(bs, 2H), 1.11-1.16(m, 2H), 0.79-0.84(m, 2H); LCMS: 540.3 [M+H]

[0389] Example 50 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-chlorophenyl)-3-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.026 g, 18% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-chlorophenyl)carbamate (C10, 0.100 g, 0.238 mmol) and 4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D1, 0.068 g, 0.238 mmol). 1 H NMR (400MHz, DMSO-d6)δ=11.14(bs, 1H), 9.76(bs, 1H), 8.17(s, 1H), 8.00-8.07(m, 2H), 7.66-7.69(m, 2H), 7.52(d, J=2.0Hz, 1H), 7.36-7.38(m, 1H), 7.32(s, 1H), 6.13(bs, 2H), 3.56-3.61(m, 1H), 3.53(s, 2H), 2.29-2.35(m, 2H), 1.73(bs, 8H), 0.97-1.08(m, 7H); LCMS: 613.3 [M+H]

[0390] Example 51 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[4,3-C]pyridin-3-yl)-2-fluorophenyl)-3-(3-methyl-4-morpholinophenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 39, starting from 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-N-(2,4-dimethoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (B11, 0.100 g, 0.231 mmol) and phenyl(3-methyl-4-morpholinophenyl)carbamate (C4, 0.072 g, 0.231 mmol) in Step 1, and obtained as an off-white solid (0.005 g, 4.3% yield over two steps). 1 H NMR(400MHz, CD3OD)δ=8.33-8.38(m, 1H), 7.72(d, J=7.2Hz, 1H), 7.44-7.52(m, 2H), 7.34(d, J=7.2Hz, 1H), 7.28-7.31(m, 2H), 7.06-7.09(m, 1H), 3.85-3.87(m, 5H), 2.91-2.93(m, 4H), 2.35(s, 3H), 1.27-1.32(m, 4H); LCMS: 502.2 [M+H]

[0391] Example 52 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.010 g, 10% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((4-methylpiperazin-1-yl)methyl)aniline (D28, 0.038 g, 0.186 mmol). 1H NMR(400MHz, CD3OD)δ=8.31-8.36(m, 2H), 7.50-7.58(m, 4H), 7.37(d, J=8.4Hz, 2H), 6.90(s, 1H), 3.82(s, 2H), 3.61-3.64(m, 1H), 3.32-3.33(m, 4H), 2.87-2.93(m, 7H), 1.11-1.16(m, 2H), 0.80-0.84(m, 2H); LCMS: 515.3 [M+H]

[0392] Example 53 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-ethylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.020 g, 20% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((4-ethylpiperazin-1-yl)methyl)aniline (D29, 0.041 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.31-8.37(m, 2H), 7.50-7.58(m, 4H), 7.37(d, J=8.4Hz, 2H), 6.90(s, 1H), 3.81(s, 2H), 3.61-3.64(m, 1H), 3.15-3.33(m, 6H), 2.97(bs, 4H), 1.31-1.37(m, 3H), 1.11-1.16(m, 2H), 0.80-0.84(m, 2H); LCMS: 529.3 [M+H]

[0393] Example 54 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(difluoromethyl)-4-((4-methylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.031 g, 20% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.077 g, 0.192 mmol) and 3-(difluoromethyl)-4-((4-methylpiperazin-1-yl)methyl)aniline (D30, 0.070 g, 0.274 mmol). 1 H NMR(400MHz, CD3OD)δ=8.26(t, J=8.4Hz, 1H), 8.16(s, 1H), 7.79(s, 1H), 7.59(d, J=8.4Hz, 1H), 7.47-7.53(m, 2H), 7.13-7.37(m, 2H), 6.64(s, 1H), 3.67(s, 2H), 3.49-3.50(m, 1H), 2.88(bs, 4H), 2.59(bs, 7H), 1.08-1.11(m, 2H), 0.80-0.84(m, 2H); LCMS: 565.5 [M+H]

[0394] Example 55 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(difluoromethyl)-4-((4-ethylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.005 g, 4.8% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 3-(difluoromethyl)-4-((4-ethylpiperazin-1-yl)methyl)aniline (D31, 0.050 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.31-8.36(m, 2H), 7.83(d, J=2.0Hz, 1H), 7.50-7.62(m, 3H), 7.12-7.40(m, 2H), 6.90(s, 1H), 3.75(s, 2H), 3.49-3.64(m, 3H), 3.19-3.24(m, 2H), 3.02(bs, 4H), 2.49(bs, 2H), 1.34-1.40(m, 3H), 1.11-1.16(m, 2H), 0.79-0.84(m, 2H); LCMS: 579.2 [M+H]

[0395] Example 56 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(6-((1-methylpiperidin-4-yl)oxy)-5-(trifluoromethyl)pyridin-3-yl)urea [ka] The title compound was obtained as an off-white solid (0.008 g, 11% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.050 g, 0.124 mmol) and 6-((1-methylpiperidin-4-yl)oxy)-5-(trifluoromethyl)pyridin-3-amine (D32, 0.034 g, 0.124 mmol). 1H NMR(400MHz, CD3OD)δ=8.44-8.45(m, 1H), 8.29-8.35(m, 3H), 7.51-7.59(m, 2H), 6.87(s, 1H), 5.40-5.60(m, 1H), 3.50-3.62(m, 5H), 2.96(s, 3H), 2.35-2.50(m, 2H), 2.18(bs, 1H), 1.98(bs, 1H), 1.10-1.13(m, 2H), 0.78-0.82(m, 2H); LCMS: 585.2 [M+H]

[0396] Example 57 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-cyclopropylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.017 g, 15% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((4-cyclopropylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D33, 0.056 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.14-8.19(m, 2H), 7.91(d, J=2.0Hz, 1H), 7.71(d, J=8.4Hz, 1H), 7.62-7.65(m, 1H), 7.25-7.30(m, 2H), 7.19(s, 1H), 3.64(s, 2H), 3.48-3.53(m, 1H), 2.75(bs, 4H), 2.52(bs, 4H), 1.75-1.99(m, 1H), 1.05-1.18(m, 4H), 0.47-0.56(m, 4H); LCMS: 609.2 [M+H]

[0397] Example 58 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-isopropylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.006 g, 5.3% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((4-isopropylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D34, 0.056 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.37(s, 1H), 8.21(t, J=8.0Hz, 1H), 7.96(d, J=2.0Hz, 1H), 7.67-7.74(m, 2H), 7.50(s, 1H), 7.29-7.37(m, 2H), 3.70-3.77(m, 3H), 3.48-3.55(m, 3H), 3.11-3.18(m, 4H), 2.53(bs, 2H), 1.40(d, J=6.4Hz, 6H), 1.18-1.23(m, 4H); LCMS: 611.3 [M+H]

[0398] Example 59 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((5-methyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.002 g, 1.8% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((5-methyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)-3-(trifluoromethyl)aniline (D35, 0.053 g, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.37(s, 1H), 8.21(t, J=8.4Hz, 1H), 7.95(d, J=2.0Hz, 1H), 7.66-7.75(m, 2H), 7.49(s, 1H), 7.29-7.37(m, 2H), 4.19(s, 1H), 3.89-4.02(m, 2H), 3.68-3.74(m, 4H), 3.37(s, 2H), 2.94(bs, 5H), 1.19-1.19(m, 4H); LCMS: 595.2 [M+H]

[0399] Example 60 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(2-fluoro-4-((4-methylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.009 g, 9.5% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 2-fluoro-4-((4-methylpiperazin-1-yl)methyl)aniline (D36, 0.041 g, 0.186 mmol). 1H NMR(400MHz, CD3OD)δ=8.36(s, 1H), 8.27(t, J=8.0Hz, 1H), 8.13(t, J=8.4Hz, 1H), 7.48(s, 1H), 7.29-7.37(m, 2H), 7.15-7.24(m, 2H), 3.66-3.73(m, 3H), 3.12(bs, 4H), 2.90(s, 3H), 2.51(bs, 4H), 1.18-1.22(m, 4H); LCMS: 533.3 [M+H]

[0400] Example 61 1-(4-((4,7-diazaspiro[2.5]octan-4-yl)methyl)-3-(trifluoromethyl)phenyl)-3-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)urea [ka]

[0401] Step 1: Synthesis of tert-butyl 4-(4-(3-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)ureido)-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane-7-carboxylate The title compound was obtained as an off-white solid (0.020 g, 15% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and tert-butyl 4-(4-amino-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane-7-carboxylate (D37, 0.072 g, 0.186 mmol). LCMS: 695.2 [M+H]

[0402] Step 2: Synthesis of 1-(4-((4,7-diazaspiro[2.5]octan-4-yl)methyl)-3-(trifluoromethyl)phenyl)-3-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)urea The title compound was prepared in a manner similar to that described in Step 2 of Intermediate A15, starting from tert-butyl 4-(4-(3-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)ureido)-2-(trifluoromethyl)benzyl)-4,7-diazaspiro[2.5]octane-7-carboxylate (0.022 g, 0.032 mmol) and obtained as an off-white solid (0.09 g, 48% yield). 1 H NMR(400MHz, CD3OD)δ=8.38(s, 1H), 8.22(t, J=8.4Hz, 1H), 7.95(d, J=2.0Hz, 1H), 7.62-7.63(m, 2H), 7.51(s, 1H), 7.29-7.37(m, 2H), 4.09(s, 2H), 3.70-3.72(m, 1H), 3.15-3.24(m, 4H), 3.04-3.07(m, 2H), 1.18-1.23(m, 4H), 0.80-0.92(m, 2H), 0.77-0.79(m, 2H); LCMS: 595.3 [M+H]

[0403] Example 62 1-(4-((4,7-diazaspiro[2.5]octan-4-yl)methyl)-3-(trifluoromethyl)phenyl)-3-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)urea [ka]

[0404] Step 1: Synthesis of 1-(5-bromo-3-fluoropyridin-2-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was obtained as an off-white solid (0.260 g, quantitative yield) following the general urea formation procedure (Method A) starting from phenyl (5-bromo-3-fluoropyridin-2-yl)carbamate (synthesized as described in PCT International Publication No. WO 2017 / 117393 A1, 0.163 g, 0.524 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (0.143 g, 0.524 mmol). It was subsequently purified by preparative HPLC (elution: 0.1% TFA / water and ACN). LCMS: 490.1 [M+H]

[0405] Step 2: Synthesis of tert-butyl (tert-butoxycarbonyl)(7-cyclopropyl-5-(5-fluoro-6-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate The title compound was prepared in a similar manner to that described for Intermediate B5, starting from 1-(5-bromo-3-fluoropyridin-2-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (0.280 g, 0.560 mmol) and tert-butyl (tert-butoxycarbonyl) (7-cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (prepared as described in IN 2016 / 11024887 A, 0.274 g, 0.560 mmol), and obtained as a brown solid (0.100 g, 23% yield). It was subsequently purified by preparative HPLC (elution: 10 mM NH4OAc / water and ACN). LCMS: 784.3 [M+H]

[0406] Step 3: Synthesis of 1-(5-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-3-fluoropyridin-2-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was prepared in a manner similar to that described in Step 2 of Intermediate A15, starting from tert-butyl (tert-butoxycarbonyl) (7-cyclopropyl-5-(5-fluoro-6-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (0.100 g, 0.128 mmol), and obtained as a white solid (0.063 g, 84% yield). It was subsequently purified by preparative HPLC (elution: 0.1% TFA / water and ACN). 1 H NMR (400MHz, DMSO-d6)δ=9.75(s, 1H), 9.50(s, 1H), 8.45(s, 1H), 8.30(s, 1H), 8.05(d, J=2.0Hz, 1H), 7.79-7.86(m, 2H), 7.66-7.68(m, 2H), 3.66-3.75(m, 3H), 3.39-3.42(m, 2H), 3.03(s, 4H), 2.90-2.93(m, 2H), 2.81(s, 3H), 2.33-2.36(m, 2H), 1.11-1.12(m, 4H); LCMS: 584.2 [M+H]

[0407] Example 63 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(6-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)pyridin-3-yl)urea [ka] The title compound was obtained as an off-white solid (0.005 g, 3.4% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.100 g, 0.248 mmol) and 6-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)pyridin-3-amine (D38, 0.068 g, 0.248 mmol). 1 H NMR(400MHz, CD3OD)δ=8.85(d, J=2.4Hz, 1H), 8.48(d, J=2.4Hz, 1H), 8.37(s, 1H), 8.21(t, J=8.0Hz, 1H), 7.49(s, 1H), 7.31-7.39(m, LCMS: 584.4 [M+H]

[0408] Example 64 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(dimethylamino)piperidin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.004 g, 3.5% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 1-(4-amino-2-(trifluoromethyl)benzyl)-N,N-dimethylpiperidin-4-amine (D39, 0.056 g, 0.186 mmol). 1H NMR(400MHz, CD3OD)δ=8.15-8.21(m, 2H), 7.89(d, J=2.0Hz, 1H), 7.66-7.73(m, 2H), 7.28-7.33(m, 2H), 7.21(s, 1H), 3.66(s, 2H), 3.50-3.54(m, 1H), 3.03-3.15(m, 3H), 2.82(s, 6H), 2.14-2.19(m, 2H), 2.04-2.07(m, 2H), 1.73-1.77(m, 2H), 1.07-1.17(m, 4H); LCMS: 611.3 [M+H]

[0409] Example 65 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((2,4-dimethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.014 g, 13% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((2,4-dimethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D40, 0.053 g, 0.186 mmol). 1H NMR(400MHz, CD3OD)δ=8.35(s, 1H), 8.22(t, J=8.4Hz, 1H), 7.95(d, J=2.0Hz, 1H), 7.76(d, J=8.0Hz, 1H), 7.66-7.69(m, 1H), 7.47(s, 1H), 7.29-7.37(m, 2H), 3.70(s, 2H), 3.48-3.51(m, 1H), 3.15-3.16(m, 1H), 2.89-2.92(m, 5H), 2.74-0.00(m, 2H), 2.49(bs, 2H), 1.26(d, J=6.0Hz, 3H), 1.17-1.22(m, 4H); LCMS: 597.3 [M+H]

[0410] Example 66 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 44, starting from phenyl(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.060 g, 0.149 mmol) and 4-((4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazin-1-yl)methyl)-3-(difluoromethyl)aniline (D41, 0.078 g, 0.149 mmol) in Step 1, and obtained as a white solid (0.004 g, 4.4% yield over two steps). It was subsequently purified by preparative HPLC (elution: 0.1% TFA / water and ACN). 1H NMR(400MHz, CD3OD)δ=8.37(s, 1H), 8.23(t, J=8.4Hz, 1H), 7.82(d, J=2.0Hz, 1H), 7.60(d, J=8.8Hz, 1H), 7.49(s, 1H), 7.13-7.40(m, LCMS: 595.3 [M+H]

[0411] Example 67 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-fluoro-4-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.002 g, 1.3% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.100 g, 0.248 mmol) and 3-fluoro-4-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)aniline (D42, 0.072 g, 0.248 mmol). LCMS: 601.3 [M+H]

[0412] Example 68 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-cyclopropyl-4-((4-methylpiperazin-1-yl)methyl)phenyl)urea [ka] The title compound was obtained as an off-white sticky material (0.010 g, 9.7% yield) starting from phenyl(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 3-cyclopropyl-4-((4-methylpiperazin-1-yl)methyl)aniline (D43, 0.046 g, 0.186 mmol) following the general procedure for urea formation (Method A). 1 H NMR (400MHz, DMSO-d6)δ=9.13(bs, 1H), 8.66(bs, 1H), 8.35(s, 1H), 8.24(t, J=8.4Hz, 1H), 7.47(s, 1H), 7.32-7.35(m, 1H), 7.18-7.24(m, 3H), 7.08(s, 1H), 3.55-3.62(m, 3H), 2.95-2.97(m, 4H), 2.78(bs, 4H), 2.20(bs, 1H), 2.08(s, 3H), 1.07-1.09(m, 4H), 0.94-0.96(m, 2H), 0.60-0.61(m, 2H); LCMS: 555.3 [M+H]

[0413] Example 69 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methyl-4,7-diazaspiro[2.5]octan-7-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white sticky material (0.004 g, 3.3% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((4-methyl-4,7-diazaspiro[2.5]octan-7-yl)methyl)-3-(trifluoromethyl)aniline (D44, 0.056 g, 0.186 mmol).1 H NMR(400MHz, CD3OD)δ=8.36(s, 1H), 8.22(t, J=8.4Hz, 1H), 7.97(d, J=1.6Hz, 1H), 7.69-7.73(m, 2H), 7.48(s, 1H), 7.29-7.37(m, 2H), 3.85(s, 2H), 3.69-3.73(m, 1H), 3.48-3.51(m, 2H), 2.92-2.95(m, 7H), 1.18-1.31(m, 6H), 0.80-0.93(m, 2H); LCMS: 609.3 [M+H]

[0414] Example 70 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(methylamino)piperidin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 61, starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and tert-butyl (1-(4-amino-2-(trifluoromethyl)benzyl)piperidin-4-yl)(methyl)carbamate (D45, 0.072 g, 0.186 mmol) in Step 1, and obtained as an off-white solid (HCl salt, 0.013 g, 7.2% yield over two steps). 1H NMR(400MHz, CD3OD)δ=8.38(s, 1H), 8.23(t, J=8.4Hz, 1H), 8.15(bs, 1H), 7.93-7.95(m, 1H), 7.82-7.84(m, 1H), 7.51(s, 1H), 7.31-7.39(m, 2H), 4.48(bs, 2H), 3.50-3.74(m, 4H), 3.02(bs, 2H), 2.77(s, 3H), 2.36(bs, 2H), 2.08(bs, 2H), 1.18-1.23(m, 4H); LCMS: 597.3 [M+H]

[0415] Example 71 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-aminopiperidin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 61, starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and tert-butyl (1-(4-amino-2-(trifluoromethyl)benzyl)piperidin-4-yl)carbamate (D46, 0.069 g, 0.186 mmol) in Step 1, and obtained as an off-white solid (HCl salt, 0.015 g, 14% yield over two steps). 1 H NMR(400MHz, CD3OD)δ=8.38(s, 1H), 8.23(t, J=8.4Hz, 1H), 8.17(bs, 1H), 7.95(bs, 1H), 7.82-7.84(m, 1H), 7.51(s, 1H), 7.31-7.39(m, LCMS: 583.3 [M+H]

[0416] Example 72 1-(4-(4-amino-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.104 g, 26% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C11, 0.280 g, 0.686 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.187 g, 0.686 mmol). 1 H NMR (400MHz, DMSO-d6)δ=9.59(bs, 1H), 8.93(bs, 1H), 8.39(s, 1H), 8.35(t, J=8.4Hz, 1H), 8.03(s, 1H), 7.64-7.67(m, 2H), 7.47-7.54(m, 2H), 4.42(t, J=6.0Hz, 2H), 3.86(t, J=5.6Hz, 2H), 3.66(s, 2H), 3.40-3.43(m, 2H), 2.91-3.03(m, 4H), 2.82(s, 3H), 2.34-2.37(m, 2H); LCMS: 588.3 [M+H]

[0417] Example 73 1-(4-(4-amino-1-(2-methoxyethyl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a light brown solid (0.040 g, 14% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-1-(2-methoxyethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C12, 0.200 g, 0.473 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.129 g, 0.473 mmol). 1 H NMR (400MHz, DMSO-d6)δ=9.47(bs, 1H), 8.84(bs, 1H), 8.34(t, J=8.0Hz, 1H), 8.25(s, 1H), 8.02(d, J=2.0Hz, 1H), 7.65(d, J=8.4Hz, 1H), 7.55-7.57(m, 1H), 7.45-7.51(m, 2H), 4.49(t, J=6.0Hz, 2H), 3.82(t, J=5.6Hz, 2H), 3.54(s, 2H), 3.23(s, 3H), 2.38(bs, 8H), 2.17(s, 3H); LCMS: 602.4 [M+H]

[0418] Example 74 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethoxy)phenyl)urea [ka] The title compound was obtained as an off-white sticky material (0.005 g, 9.6% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.035 g, 0.087 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethoxy)aniline (D47, 0.025 g, 0.087 mmol). 1H NMR(400MHz, CD3OD)δ=8.36(s, 1H), 8.20(t, J=8.4Hz, 1H), 7.78(s, 1H), 7.48-7.50(m, 2H), 7.29-7.37(m, 3H), 3.69-3.72(m, 3H), 3.45(bs, 2H), 3.06(bs, 4H), 2.90(s, 3H), 2.52(bs, 2H), 1.17-1.22(m, 4H); LCMS: 599.7 [M+H]

[0419] Example 75 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-(piperazin-1-ylmethyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 61, starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and tert-butyl 4-(4-amino-2-(trifluoromethyl)benzyl)piperazine-1-carboxylate (D48, 0.067 g, 0.186 mmol) in Step 1, and obtained as an off-white solid (HCl salt, 0.009 g, 8.4% yield over two steps). 1 H NMR (400MHz, DMSO-d6)δ=10.03(bs, 1H), 9.00(bs, 1H), 8.92(bs, 1H), 8.50(s, 1H), 8.19(t, J=8.4Hz, 1H), 8.05(s, 1H), 7.68(d, J=8.8Hz, 1H), 7.59(bs, 2H), 7.24-7.39(m, 2H), 3.70-3.73(m, 3H), 3.13(bs, 4H), 2.68(bs, 4H), 1.10-1.13(m, 4H); LCMS: 569.2 [M+H]

[0420] Example 76 1-(4-((4-acetylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)-3-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)urea [ka] The title compound was obtained as an off-white solid (0.013 g, 11% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 1-(4-(4-amino-2-(trifluoromethyl)benzyl)piperazin-1-yl)ethan-1-one (D49, 0.056 g, 0.186 mmol). 1 H NMR (400MHz, DMSO-d6)δ=9.40(bs, 1H), 8.68(bs, 1H), 8.18-8.23(m, 2H), 8.02(d, J=2.0Hz, 1H), 7.69(d, J=8.4Hz, 1H), 7.57(d, J=8.4Hz, LCMS: 611.4 [M+H]

[0421] Example 77 1-(4-(4-amino-7-(2-methoxyethyl)-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a white solid (0.08 g, 19% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C13, 0.290 g, 0.688 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.188 g, 0.688 mmol). 1 H NMR(400MHz, CD3OD)δ=8.35(s, 1H), 8.21-8.23(m, 1H), 7.96(d, J=2.0Hz, 1H), 7.67-7.73(m, 2H), 7.56(s, 1H), 7.31-7.39(m, 2H), 4.54(t, J=5.2Hz, 2H), 3.82(t, J=4.8Hz, 2H), 3.76(s, 2H), 3.49-3.51(m, 2H), 3.37(s, 3H), 3.14-3.19(m, 2H), 3.04-3.06(m, 2H), 2.93(s, 3H), 2.47(bs, 2H); LCMS: 601.5 [M+H]

[0422] Example 78 1-(4-((1-acetylpiperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)-3-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)urea [ka] The title compound was obtained as an off-white solid (0.04 g, 33% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.08 g, 0.198 mmol) and 1-(4-(4-amino-2-(trifluoromethyl)benzyl)piperazin-1-yl)ethan-1-one (D50, 0.06 g, 0.198 mmol). 1H NMR(400MHz, CD3OD)δ=8.36(s, 1H), 8.20(t, J=8.4Hz, 1H), 7.82(d, J=2.4Hz, 1H), 7.59-7.62(m, 1H), 7.48(s, 1H), 7.22-7.36(m, 3H), 4.82-4.84(m, 1H), 3.58-3.80(m, 5H), 2.15(s, 3H), 1.89(bs, 4H), 1.18-1.22(m, 4H); LCMS: 612.2 [M+H]

[0423] Example 79 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((1-(oxetan-3-yl)piperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as a white solid (0.058 g, 62% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.060 g, 0.149 mmol) and 4-((1-(oxetan-3-yl)piperidin-4-yl)oxy)-3-(trifluoromethyl)aniline (D51, 0.047 g, 0.149 mmol). 1H NMR (400MHz, DMSO-d6)δ=9.29(bs, 1H), 8.73(bs, 1H), 8.40(s, 1H), 8.23(t, J=8.4Hz, 1H), 7.92(s, 1H), 7.56-7.59(m, 1H), 7.54(s, 1H), 7.35-7.38(m, 2H), 7.26(d, J=8.4Hz, 1H), 4.70-4.78(m, 5H), 4.38-4.41(m, 1H), 3.66-3.72(m, 1H), 2.83-2.98(m, 4H), 2.12(bs, 3H), 1.83(bs, 1H), 1.06-1.11(m, 4H); LCMS: 626.2 [M+H]

[0424] Example 80 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(2-fluoroethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.026 g, 23% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((4-(2-fluoroethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D52, 0.057 g, 0.186 mmol). 1H NMR (400MHz, DMSO-d6)δ=9.38(bs, 1H), 8.67(bs, 1H), 8.14-8.22(m, 2H), 8.01(d, J=2.4Hz, 1H), 7.66(d, J=8.4Hz, 1H), 7.54-7.57(m, 1H), 7.29-7.34(m, 2H), 7.23-7.26(m, 1H), 6.15(bs, 2H), 4.59(t, J=4.8Hz, 1H), 4.47(t, J=5.2Hz, 1H), 3.55-3.60(m, 3H), 2.66-2.67(m, 2H), 2.40(bs, 2H), 2.08(bs, 6H), 1.02-1.06(m, 4H); LCMS: 615.3 [M+H]

[0425] Example 81 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4,4-difluoropiperidin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.006 g, 8.3% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.050 g, 0.124 mmol) and 4-((4,4-difluoropiperidin-1-yl)methyl)-3-(trifluoromethyl)aniline (D53, 0.036 g, 0.124 mmol). 1H NMR(400MHz, CD3OD)δ=8.16-8.20(m, 2H), 7.92(d, J=2.0Hz, 1H), 7.73(d, J=8.4Hz, 1H), 7.63-7.66(m, 1H), 7.26-7.32(m, 2H), 7.21(s, LCMS: 604.3 [M+H]

[0426] Example 82 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(oxetan-3-yl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.023 g, 20% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((4-(oxetan-3-yl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D54, 0.059 g, 0.186 mmol). 1 H NMR (400MHz, DMSO-d6)δ=10.88(bs, 1H), 10.05(bs, 1H), 8.16(d, J=3.6Hz, 1H), 7.99-8.06(m, 2H), 7.58-7.66(m, 2H), 7.20-7.30(m, 3H), 6.13(bs, 2H), 4.52(t, J=6.4Hz, 2H), 4.42(t, J=6.0Hz, 2H), 3.54-3.60(m, 3H), 2.42(bs, 4H), 2.28-2.34(m, 4H), 1.02-1.07(m, 4H); LCMS: 625.3 [M+H]

[0427] Example 83 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(3-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.005 g, 3.5% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C5, 0.100 g, 0.247 mmol) and 3-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)aniline (D55, 0.068 g, 0.247 mmol). 1 H NMR(400MHz, CD3OD)δ=8.29-8.36(m, 2H), 7.84(s, 1H), 7.69(s, 1H), 7.47-7.52(m, 2H), 7.34(s, 1H), 3.80-3.84(m, 1H), 3.64(s, 2H), 2.77(bs, 4H), 2.64(bs, 4H), 2.49(s, 3H), 1.29-1.33(m, 2H), 1.18-1.21(m, 2H); LCMS: 584.3 [M+H]

[0428] Example 84 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((1-ethylpiperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.026 g, 23% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((1-ethylpiperidin-4-yl)oxy)-3-(trifluoromethyl)aniline (D56, 0.054 g, 0.186 mmol). 1 H NMR (400MHz, DMSO-d6)δ=9.30(bs, 1H), 8.73(bs, 1H), 8.39(s, 1H), 8.22(t, J=8.4Hz, 1H), 7.89-7.93(m, 1H), 7.50-7.59(m, 2H), 7.24-7.37(m, 3H), 4.66-4.93(m, 1H), 3.45-3.71(m, 3H), 3.13-3.17(m, 2H), 2.91-3.06(m, 2H), 2.27-2.33(m, 1H), 2.04-2.13(m, 2H), 1.76-1.79(m, 1H), 1.24(t, J=7.2Hz, 3H), 1.09-1.10(m, 4H); LCMS: 598.4 [M+H]

[0429] Examples 85 and 86 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((3-fluoro-1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)urea [ka] The two diastereomers of the title compound were obtained following the general urea formation procedure (Method A). Example 85 was prepared using phenyl(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.070 g, 0.174 mmol) and one diastereomer of 4-((3-fluoro-1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)aniline (D57a, 0.051 g, 0.174 mmol) and obtained as an off-white solid (0.010 g, 9.5% yield). 1 H NMR(400MHz, CD3OD)δ=8.14-8.20(m, 2H), 7.79(d, J=2.8Hz, 1H), 7.60-7.63(m, 1H), 7.20-7.31(m, 4H), 4.60-4.84(m, 2H), 3.49-3.54(m, 1H), 2.97(bs, 1H), 2.70-2.72(m, 2H), 2.50(bs, 1H), 2.40(s, 3H), 2.13-2.15(m, 1H), 1.94-1.99(m, 1H), 1.06-1.18(m, 4H); LCMS: 602.3 [M+H]

[0430] Example 86 was prepared using phenyl(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and the other diastereomer of 4-((3-fluoro-1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)aniline (D57b, 0.054 g, 0.186 mmol) and obtained as an off-white solid (0.007 g, 6% yield). 1H NMR (400MHz, DMSO-d6)δ=9.25(bs, 1H), 8.66(bs, 1H), 8.14-8.19(m, 2H), 7.87(d, J=2.4Hz, 1H), 7.52-7.55(m, 1H), 7.22-7.37(m, 4H), 6.13(bs, 2H), 4.56-4.71(m, 2H), 2.94-3.00(m, 1H), 2.59-2.68(m, 1H), 2.04-2.33(m, 7H), 1.58-1.61(m, 1H), 1.02-1.04(m, 4H); LCMS: 602.3 [M+H]

[0431] Examples 87 and 88 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((3-fluoro-1-(oxetan-3-yl)piperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)urea [ka] The two diastereomers of the title compound were obtained following the general urea formation procedure (Method A). Example 87 was prepared using phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.050 g, 0.124 mmol) and one diastereomer of 4-((3-fluoro-1-(oxetan-3-yl)piperidin-4-yl)oxy)-3-(trifluoromethyl)aniline (D58a, 0.041 g, 0.124 mmol) and obtained as an off-white solid (0.005 g, 6.3% yield). 1H NMR(400MHz, CD3OD)δ=8.14-8.20(m, 2H), 7.79(d, J=2.8Hz, 1H), 7.59-7.62(m, 1H), 7.21-7.31(m, 4H), 4.60-4.88(m, 6H), 3.64-3.67(m, 1H), 3.50-3.54(m, 1H), 2.42-2.76(m, 4H), 1.92-2.18(m, 2H), 1.06-1.18(m, 4H); LCMS: 644.3 [M+H] Example 88 was prepared using phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and the other diastereomer of 4-((3-fluoro-1-(oxetan-3-yl)piperidin-4-yl)oxy)-3-(trifluoromethyl)aniline (D58b, 0.062 g, 0.186 mmol) and obtained as an off-white solid (0.006 g, 5.4% yield). 1 H NMR (400MHz, DMSO-d6)δ=9.21(bs, 1H), 8.63(bs, 1H), 8.17-8.21(m, 2H), 7.88(d, J=2.4Hz, 1H), 7.52-7.55(m, 1H), 7.22-7.38(m, 4H), 6.15(bs, 2H), 4.41-4.56(m, 7H), 3.51-3.59(m, 2H), 2.92-2.97(m, 1H), 2.33-2.34(m, 1H), 2.08-2.13(m, 2H), 1.60-1.62(m, 1H), 1.02-1.07(m, 4H); LCMS: 644.3 [M+H]

[0432] Example 89 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.025 g, 17% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.100 g, 0.248 mmol) and 3-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)aniline (D55, 0.068 g, 0.248 mmol). 1 H NMR(400MHz, CD3OD)δ=8.15-8.20(m, 2H), 7.82(s, 1H), 7.65(s, 1H), 7.20-7.33(m, 4H), 3.64(s, 2H), 3.50-3.53(m, 1H), 2.81(bs, 4H), 2.64-2.68(m, 4H), 2.51(s, 3H), 1.08-1.16(m, 4H); LCMS: 583.3 [M+H]

[0433] Example 90 1-(4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(3,3-difluoro-1-(4-methylpiperazin-1-yl)-2,3-dihydro-1H-inden-5-yl)urea [ka] The title compound was obtained as an off-white solid (0.010 g, 11% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C5, 0.065 g, 0.161 mmol) and 3,3-difluoro-1-(4-methylpiperazin-1-yl)-2,3-dihydro-1H-inden-5-amine (D59, 0.043 g, 0.161 mmol). 1H NMR(400MHz, CD3OD)δ=8.29-8.35(m, 2H), 7.81(s, 1H), 7.58-7.60(m, 1H), 7.46-7.52(m, 3H), 3.78-3.84(m, 1H), 2.63-2.83(m, 9H), 2.52-2.58(m, 5H), 1.18-1.33(m, 4H); LCMS: 578.3 [M+H]

[0434] Example 91 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-(piperidin-1-ylmethyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.018 g, 15% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.083 g, 0.205 mmol) and 4-(piperidin-1-ylmethyl)-3-(trifluoromethyl)aniline (D60, 0.061 g, 0.205 mmol). 1 H NMR(400MHz, CD3OD)δ=8.16-8.20(m, 2H), 7.94(d, J=2.0Hz, 1H), 7.75(d, J=8.4Hz, 1H), 7.64-7.66(m, 1H), 7.27-7.32(m, 2H), 7.21(s, 1H), 3.68(bs, 2H), 3.49-3.55(m, 1H), 2.54(bs, 4H), 1.53-1.69(m, 6H), 1.09-1.18(m, 4H); LCMS: 568.2 [M+H]

[0435] Example 92 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-hydroxypiperidin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.015 g, 12% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.085 g, 0.211 mmol) and 1-(4-amino-2-(trifluoromethyl)benzyl)piperidin-4-ol (D61, 0.058 g, 0.211 mmol). 1 H NMR(400MHz, CD3OD)δ=8.16-8.20(m, 2H), 7.94(d, J=2.0Hz, 1H), 7.74(d, J=8.4Hz, 1H), 7.64-7.67(m, 1H), 7.27-7.32(m, 2H), 7.21(s, 1H), 3.69-3.71(m, 3H), 3.50-3.54(m, 1H), 2.86-2.88(m, 2H), 2.29-2.34(m, 2H), 1.87-1.96(m, 2H), 1.60-1.64(m, 2H), 1.07-1.18(m, 4H); LCMS: 584.2 [M+H]

[0436] Example 93 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(cyclopropylsulfonyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.016 g, 9.3% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.10 g, 0.248 mmol) and 4-((4-(cyclopropylsulfonyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D62, 0.09 g, 0.248 mmol). 1 H NMR(400MHz, CD3OD)δ=7.92-8.20(m, 2H), 7.92(d, J=2.0Hz, 1H), 7.73(d, J=8.4Hz, 1H), 7.67(d, J=2.0Hz, 1H), 7.27-7.32(m, 2H), 7.21(s, 1H), 3.69(s, 2H), 3.51-3.53(m, 1H), 2.49-2.59(m, 9H), 1.03-1.16(m, 8H); LCMS: 673.3 [M+H]

[0437] Example 94 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(methylsulfonyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.002 g, 1.9% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.065 g, 0.161 mmol) and 4-((4-(methylsulfonyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D63, 0.054 g, 0.161 mmol). 1H NMR (400MHz, CD3OD) δ=8.37(s, 1H), 8.22(t, J=8.4Hz, 1H), 8.11(d, J=2.0Hz, 1H), 7.76-7.79(m, 2H), 7.50(s, 1H), 7.30-7.38(m, 2H), LCMS: 647.1 [M+H]

[0438] Example 95 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.005 g, 4% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.075 g, 0.186 mmol) and 4-((4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D64, 0.068 mg, 0.186 mmol). 1 H NMR(400MHz, CD3OD)δ=8.36(s, 1H), 8.21(t, J=8.4Hz, 1H), 8.07(bs, 1H), 7.72-7.80(m, 2H), 7.48(s, 1H), 7.30-7.38(m, 2H), 4.23(bs, LCMS: 675.5 [M+H]

[0439] Example 96 2-(4-(4-(3-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)ureido)-2-(trifluoromethyl)benzyl)piperazin-1-yl)ethane-1-sulfonic acid [ka] The title compound was obtained as an off-white solid (0.003 g, 3.6% yield) following the general urea formation procedure (Method A), starting from phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C6, 0.050 g, 0.124 mmol) and 2-(4-(4-amino-2-(trifluoromethyl)benzyl)piperazin-1-yl)ethane-1-sulfonic acid (D65, 0.046 g, 0.124 mmol). 1 H NMR(400MHz, CD3OD)δ=8.37(s, 1H), 8.22(t, J=8.0Hz, 1H), 7.97(d, J=2.0Hz, 1H), 7.66-7.74(m, 2H), 7.50(s, 1H), 7.29-7.38(m, 2H), 3.70-3.91(m, 3H), 3.51-3.69(m, 4H), 3.21-3.25(m, 2H), 3.08(bs, 4H), 2.60(bs, 2H), 1.18-1.23(m, 4H); LCMS: 677.1 [M+H]

[0440] Example 97 1-(4-(4-amino-7-(1-(methylsulfonyl)piperidin-4-yl)-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.003 g, 33% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-(1-(methylsulfonyl)piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C14, 0.02 g, 0.038 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.01 g, 0.038 mmol). 1 H NMR(400MHz, CD3OD)δ=8.17-8.21(m, 2H), 7.92(d, J=2.0Hz, 1H), 7.65-7.73(m, 2H), 7.41(s, 1H), 7.29-7.35(m, 2H), 4.79-4.82(m, 1H), 3.94-3.97(m, 2H), 3.68(s, 2H), 3.01-3.08(m, 2H), 2.94(s, 3H), 2.77(bs, 4H), 2.61(bs, 4H), 2.50(s, 3H), 2.18-2.24(m, 4H); LCMS: 704.3 [M+H]

[0441] Example 98 1-(4-(4-amino-7-(1-(cyclopropylsulfonyl)piperidin-4-yl)-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.011 g, 14% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-(1-(methylsulfonyl)piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C15, 0.06 g, 0.109 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.03 g, 0.109 mmol). 1H NMR(400MHz, CD3OD)δ=8.36(s, 1H), 8.23(t, J=8.0Hz, 1H), 7.96(d, J=4.0Hz, 1H), 7.69-7.73(m, 3H), 7.32-7.40(m, 2H), 4.92-4.96(m, 1H), 3.97-4.00(m, 2H), 3.76(s, 2H), 3.49-3.51(m, 1H), 3.14-3.18(m, 6H), 2.93(s, 3H), 2.56-2.60(m, 4H), 2.17-2.32(m, 4H), 1.08-1.12(m, 4H); LCMS: 730.3 [M+H]

[0442] Example 99 1-(4-(4-amino-1-(5-hydroxypentyl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.003 g, 5.2% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-7-(1-(methylsulfonyl)piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (C16, 0.042 g, 0.093 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.025 g, 0.093 mmol). LCMS: 630.5 [M+H]

[0443] Example 100 1-(4-(4-amino-7-(1-methylazetidin-3-yl)-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka]

[0444] Step 1: Synthesis of 1-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was obtained as an off-white solid (0.120 g, 18% yield) following the general urea formation procedure (Method A) starting from 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.300 g, 1.265 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.498 g, 1.265 mmol). LCMS: 537.3 [M+H]

[0445] Step 2: Synthesis of 1-(4-(4-amino-7-(1-methylazetidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea The title compound was prepared in a manner similar to that described for Intermediate B5, starting from 5-iodo-7-(1-methylazetidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A18, 0.035 g, 0.106 mmol) and 1-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (0.057 g, 0.106 mmol), and obtained as an off-white solid (0.014 g, 22% yield). 1H NMR (400MHz, DMSO-d6)δ=9.99(bs, 1H), 9.52(bs, 1H), 8.81(s, 1H), 8.24-8.32(m, 2H), 8.03(d, J=2.0Hz, 1H), 7.59-7.66(m, 2H), 7.10-7.37(m, 2H), 6.97(bs, 2H), 5.60-5.72(m, 1H), 4.49-4.82(m, 4H), 3.65(bs, 2H), 3.40-3.43(m, 2H), 3.03-3.07(m, 4H), 2.89-2.92(m, 2H), 2.82(s, 3H), 2.51(s, 3H); LCMS: 612.3 [M+H]

[0446] Example 101 1-(4-(4-amino-7-isopropyl-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 100, starting from 5-iodo-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (A19, 0.035 g, 0.116 mmol) and 1-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (Step 1 of Example 100, 0.062 g, 0.116 mmol), and obtained as an off-white solid (0.005 g, 7.4% yield). 1H NMR(400MHz, CD3OD)δ=8.17-8.21(m, 2H), 7.92(d, J=2.0Hz, 1H), 7.63-7.73(m, 2H), 7.38(s, 1H), 7.29-7.34(m, 2H), 5.01-5.06(m, 1H), 3.66(s, 2H), 2.58(bs, 8H), 2.37(s, 3H), 1.56(d, J=6.8Hz, 6H); LCMS: 585.3 [M+H]

[0447] Example 102 3-(4-amino-5-(3-fluoro-4-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)phenyl)-7H-pyrrolo[2,3-D]pyrimidin-7-yl)propanoic acid [ka]

[0448] Step 1: Synthesis of tert-butyl 3-(4-amino-5-(3-fluoro-4-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propanoate The title compound was prepared in a similar manner as described for Intermediate B5, starting from tert-butyl 3-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propanoate (A20, 0.045 g, 0.116 mmol) and 1-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (Step 1 of Example 100, 0.062 g, 0.116 mmol), and obtained as a white solid (0.020 g, 26% yield). LCMS: 671.3 [M+H]

[0449] Step 2: Synthesis of 3-(4-amino-5-(3-fluoro-4-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propanoic acid The title compound was prepared in a manner similar to that described in Step 3 of Example 32, starting from tert-butyl 3-(4-amino-5-(3-fluoro-4-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propanoate (0.020 g, 0.030 mmol) as an off-white solid (0.002 g, 11% yield). LCMS: 615.4 [M+H]

[0450] Example 103 4-(4-amino-5-(3-fluoro-4-(3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)ureido)phenyl)-7H-pyrrolo[2,3-D]pyrimidin-7-yl)butanoic acid [ka] The title compound was prepared in a manner similar to that described in Example 102, starting from tert-butyl 4-(4-amino-5-(3-fluoro-4-((phenoxycarbonyl)amino)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)butanoate (C17, 0.050 g, 0.099 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.027 g, 0.099 mmol) in Step 1, and obtained as an off-white solid (0.009 g, 14% yield over two steps). 1H NMR (400MHz, DMSO-d6)δ=10.00(bs, 1H), 8.90(bs, 1H), 8.35(s, 1H), 8.24(t, J=8.8Hz, 1H), 8.01(s, 1H), 7.59-7.66(m, 3H), 7.34-7.37(m, 1H), 7.15-7.27(m, 1H), 4.25(t, J=7.2Hz, 2H), 3.64(s, 2H), 3.37-3.49(m, 2H), 3.00-3.03(m, 2H), 2.88-2.91(m, 2H), 2.79(s, 3H), 2.22-2.38(m, 4H), 2.02-2.04(m, 2H); LCMS: 629.3 [M+H]

[0451] Example 104 1-(4-(4-amino-1-(1-methylpyrrolidin-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a similar manner as described in Example 100, starting from 3-iodo-1-(1-methylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (A22, 0.030 g, 0.087 mmol) and 1-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (Step 1 of Example 100, 0.051 g, 0.096 mmol), and obtained as an off-white solid (0.002 g, 3.6% yield). LCMS: 627.2 [M+H]

[0452] Example 105 1-(4-(4-amino-1-(piperidin-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 61, starting from tert-butyl 3-(4-amino-3-(3-fluoro-4-((phenoxycarbonyl)amino)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylate (C18, 0.434 g, 0.793 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.217 g, 0.793 mmol) in Step 1, and obtained as an off-white solid (0.012 g, 30% yield over two steps). It was subsequently purified by preparative HPLC (elution: 10 mM NaHCO3 / water and ACN). 1 H NMR (400MHz, DMSO-d6)δ=9.54(bs, 1H), 8.90(bs, 1H), 8.25-8.35(m, 2H), 8.03(d, J=2.0Hz, 1H), 7.66(d, J=8.4Hz, 1H), 7.56-7.58(m, 1H), 7.44-7.51(m, 2H), 6.95(bs, 2H), 4.68-4.75(m, 1H), 3.55(s, 2H), 2.94-3.14(m, 4H), 2.52-2.55(m, 2H), 2.34-2.42(m, 4H), 2.17(s, 3H), 2.04-2.13(m, 4H), 1.77-1.79(m, 1H), 1.58-1.61(m, 1H); LCMS: 627.2 [M+H]

[0453] Example 106 1-(4-(4-amino-1-(1-methylpiperidin-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Step 3 of Intermediate D17, starting from 1-(4-(4-amino-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (Example 105, 0.10 g, 0.160 mmol), and obtained as an off-white solid (0.037 g, 36% yield). It was subsequently purified by preparative HPLC (elution: 10 mM NaHCO / water and ACN). 1 H NMR (400MHz, DMSO-d6)δ=9.62(bs, 1H), 8.97(bs, 1H), 8.32(t, J=8.4Hz, 1H), 8.25(s, 1H), 8.03(d, J=2.0Hz, 1H), 7.66(d, J=8.8Hz, 1H), 7.56-7.58(m, 1H), 7.43-7.49(m, 2H), 6.94(bs, 2H), 4.75-4.81(m, 1H), 3.54(s, 2H), 2.92-2.96(m, 1H), 2.79-2.81(m, 1H), 2.33-2.46(m, 8H), 2.23(s, LCMS: 641.3 [M+H]

[0454] Example 107 1-(4-(4-amino-1-(1-methylpiperidin-4-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 100, starting from 3-iodo-1-(1-methylpiperidin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (A24, 0.12 g, 0.335 mmol) and 1-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (Step 1 of Example 100, 0.18 g, 0.335 mmol), and obtained as an off-white solid (0.04 g, 19% yield). 1 H NMR (400MHz, DMSO-d6)δ=9.47(bs, 1H), 8.84(bs, 1H), 8.34(t, J=8.4Hz, 1H), 8.26(s, 1H), 8.02(s, 1H), 7.66(d, J=8.4Hz, 1H), 7.55-7.57(m, 1H), 7.45-7.50(m, 2H), 7.00(bs, 2H), 4.61-4.67(m, 1H), 3.55(s, 2H), 2.90-2.93(m, 2H), 2.34-2.39(m, 8H), 2.21-2.24(m, 5H), 2.16(s, 3H), 2.06-2.12(m, 2H), 1.87-1.89(m, 2H); LCMS: 641.2 [M+H]

[0455] Example 108 1-(4-(4-amino-1-(azetidin-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 61, starting from tert-butyl 3-(4-amino-3-(3-fluoro-4-((phenoxycarbonyl)amino)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)azetidine-1-carboxylate (C19, 0.400 g, 0.770 mmol) and 4-((1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)aniline (D12, 0.211 g, 0.770 mmol) in Step 1, and obtained as an off-white solid (0.013 g, 20% yield over two steps). It was subsequently purified by preparative HPLC (elution: 10 mM NaHCO3 / water and ACN). 1 H NMR (400MHz, DMSO-d6)δ=9.51(bs, 1H), 8.88(bs, 1H), 8.38(t, J=8.4Hz, 1H), 8.29(s, 1H), 7.90(d, J=2.4Hz, 1H), 7.47-7.58(m, 3H), 7.28-7.30(m, 1H), 5.80-5.84(m, 1H), 4.82(bs, 1H), 4.44-4.57(m, 4H), 3.02(bs, 4H), 2.56(s, 3H), 2.09(bs, 2H), 1.88-1.90(m, 2H); LCMS: 600.2 [M+H]

[0456] Example 109 1-(4-(4-amino-1-(1-methylazetidin-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Step 3 of Intermediate D17, starting from 1-(4-(4-amino-1-(azetidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)urea (Example 108, 0.075 g, 0.125 mmol), and obtained as an off-white solid (0.003 g, 3.9% yield). It was subsequently purified by preparative HPLC (elution: 10 mM NaHCO / water and ACN). 1 H NMR(400MHz, CD3OD)δ=8.33(t, J=8.0Hz, 1H), 8.27(s, 1H), 7.80(d, J=2.8Hz, 1H), 7.52-7.63(m, 3H), 7.21(d, J=8.8Hz, 1H), 5.56-5.64(m, 1H), 4.82(bs, 1H), 4.03-4.07(m, 2H), 3.95-3.99(m, 2H), 2.82(bs, 2H), 2.62-2.68(m, 5H), 2.44(s, 3H), 1.94-2.10(m, 4H); LCMS: 614.3 [M+H]

[0457] Example 110 1-(4-(4-amino-1-(1-methylazetidin-3-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was prepared in a manner similar to that described in Example 100, starting with 3-iodo-1-(1-methylazetidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (A26, 0.130 g, 0.394 mmol) and 1-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea (Step 1 of Example 100, 0.211 g, 0.394 mmol), and obtained as an off-white solid (0.02 g, 8.3% yield). 1 H NMR (400MHz, DMSO-d6)δ=9.44(bs, 1H), 8.82(bs, 1H), 8.36(bs, 1H), 8.25(s, 1H), 8.02(s, 1H), 7.65-7.68(m, 1H), 7.47-7.57(m, 3H), 7.10(bs, 2H), 5.38(bs, 1H), 3.80(bs, 2H), 3.55-3.58(m, 4H), 2.38(bs, 8H), 2.17(bs, 6H); LCMS: 613.3 [M+H]

[0458] Example 111 1-(4-(4-amino-1-(prop-2-yn-1-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.023 g, 34% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-1-(prop-2-yn-1-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C20, 0.061 g, 0.152 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.041 g, 0.152 mmol). 1H NMR(400MHz, CD3OD)δ=8.36(t, J=8.4Hz, 1H), 8.31(s, 1H), 7.93(d, J=2.0Hz, 1H), 7.72(d, J=8.4Hz, 1H), 7.65-7.67(m, 1H), 7.51-7.57(m, 2H), 5.24(s, 2H), 3.67(s, 2H), 2.85(s, 1H), 2.70(bs, 4H), 2.59(bs, 4H), 2.44(s, 3H); LCMS: 582.1 [M+H]

[0459] Example 112 1-(4-(4-amino-1-(prop-2-yn-1-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.024 g, 27% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-1-(prop-2-yn-1-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C20, 0.061 g, 0.152 mmol) and 4-((1-methylpiperidin-4-yl)oxy)-3-(trifluoromethyl)aniline (D12, 0.042 g, 0.152 mmol). 1 H NMR(400MHz, CD3OD)δ=8.31-8.36(m, 2H), 7.79(d, J=2.4Hz, 1H), 7.50-7.62(m, 3H), 7.19(d, J=8.8Hz, 1H), 5.24(s, 2H), 4.62(s, 1H), 2.85(s, 1H), 2.71(bs, 2H), 2.50(bs, 2H), 2.34(s, 3H), 1.93-2.06(m, 4H); LCMS: 583.2 [M+H]

[0460] Example 113 1-(4-(4-amino-1-(but-3-yn-1-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.025 g, 23% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-1-(but-3-yn-1-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C21, 0.07 g, 0.180 mmol) and 4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (D6, 0.049 g, 0.180 mmol). 1 H NMR (400MHz, DMSO-d6)δ=9.45(s, 1H), 8.82(s, 1H), 8.35(t, J=8.4Hz, 1H), 8.26(s, 1H), 8.02(s, 1H), 7.66(d, J=8.4Hz, 1H), 7.46-7.57(m, 3H), 7.02(bs, 2H), 4.45-4.48(m, 2H), 3.55(s, 2H), 2.78-2.82(m, 3H), 2.33-2.38(m, 8H), 2.18(s, 3H); LCMS: 596.2 [M+H]

[0461] Example 114 1-(4-(4-amino-1-(but-3-yn-1-yl)-1H-pyrazolo[3,4-D]pyrimidin-3-yl)-2-fluorophenyl)-3-(4-((1-ethylpiperidin-4-yl)oxy)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.023 g, 21% yield) following the general urea formation procedure (Method A) starting from phenyl (4-(4-amino-1-(but-3-yn-1-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)carbamate (C21, 0.076 g, 0.183 mmol) and 4-((1-ethylpiperidin-4-yl)oxy)-3-(trifluoromethyl)aniline (D56, 0.053 g, 0.183 mmol). 1 H NMR (400MHz, DMSO-d6)δ=9.21(s, 1H), 8.74(s, 1H), 8.34(t, J=8.4Hz, 1H), 8.26(s, 1H), 7.87(d, J=2.4Hz, 1H), 7.45-7.54(m, 3H), 7.27-7.29(m, 1H), 7.02(bs, 2H), 4.45-4.56(m, 3H), 2.78-2.82(m, 3H), 2.60-2.67(m, 2H), 2.27-2.36(m, 4H), 1.90-1.93(m, 2H), 1.66-1.68(m, 2H), 1.00(t, J=6.80Hz, 3H); LCMS: 611.2 [M+H]

[0462] Example 115 1-(4-(4-amino-7-(2-hydroxyethyl)-7H-pyrrolo[2,3-D]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea [ka] The title compound was obtained as an off-white solid (0.007 mg, 3.4% yield) following the general urea formation procedure (Method A) starting from 2-(4-amino-5-(4-amino-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-ol (B25, 0.109 g, 0.379 mmol) and phenyl(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamate (C1, 0.149 g, 0.379 mmol).1 H NMR(400MHz, CD3OD)δ=8.14-8.18(m, 2H), 7.92(d, J=2.0Hz, 1H), 7.64-7.71(m, 2H), 7.28-7.33(m, 3H), 4.31-4.34(m, 2H), 3.93(t, LCMS: 587.3 [M+H]

[0463] Biological Example 1 NEK7 enzyme assay Casein substrate (a mixture of hydrolyzed and partially dephosphorylated α-, β-, and κ-caseins from bovine milk, purchased from Sigma-Aldrich, catalog number C4765, diluted in distilled water to a final concentration of 1 mg / mL, assay concentration 20 μM) and full-length recombinant human Nek7 (expressed by baculovirus in Sf9 insect cells with an N-terminal GST tag, purchased from SignalChem, catalog number N09-10G, 0.1 μg / μL, assay concentration 70 nM) 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 target compound (3-fold serial dilutions from 10 μM to 0.5 nM in DMSO) or control (1% DMSO) was dispensed into the kinase reaction mixture using acoustic technology (Echo550, nanoliter range). After 20 minutes of incubation at room temperature, [ 32 The kinase reaction was initiated by adding [P]-ATP (specific activity at 10 μCi / μL), and the mixture was incubated for 2 hours at room temperature. The reaction was then stopped by spotting the reaction mixture onto phosphocellulose paper (P81). After washing, the specific radioactivity of the P81 paper was measured, and kinase activity data were expressed as the percentage of kinase activity in the test sample compared to the reaction in vehicle (dimethyl sulfoxide). IC 50 Value calculation and curve fitting were performed using Prism (GraphPad Software).

[0464] Biological Example 2 IL-1β release assay Approximately 1.5 million THP-1 cells were seeded into each well of a 6-well TC plate and incubated with 40 nM PMA / RPMI (10% FBS, 1% Pen-Strep) for 24 hours. The medium was then removed, and the cells were rested in RPMI (10% FBS, 1% Pen-Strep) for 24 hours. After this, the medium was removed, and the cells were pre-treated with various concentrations of the compound of interest (typically 3-fold serial dilutions in RPMI + 5% FBS, concentrations: 1 μM to 0.5 nM) in RPMI (5% FBS) for 2 hours. The medium was removed again, and the cells were incubated with LPS (250 ng / mL) and the compound of interest (at the concentrations listed above) in RPMI (5% FBS) for 2 hours. Finally, the medium was removed, and the cells were incubated with nigericin (20 μM) and the compound of interest (at the concentrations listed above) in Opti-MEM for 30 minutes. The cell culture medium was then collected and the amount of bound IL-1β was determined using JESS (Protein Simple) and standard protocols. The bound IL-1β antibody was obtained from Cell Signaling (Cat. No. 83186S) and diluted 1:20 with Antibody Diluent 2. A 1x anti-rabbit HRP secondary antibody (Protein Simple) was used for chemiluminescence detection with Protein Simple's luminol and peroxide. Incubation time with the primary antibody was increased from 30 to 60 minutes.

[0465] Biological Example 3 Biochemical assays of representative compounds Representative compounds were tested for inhibitory activity against NEK7 and IL-1β release according to the procedures described above, and the results are shown in the table below. [Table 38] [Table 39] NEK7 IC in Table 2 50 Regarding activity: * indicates values ​​above 2,000 nM ** indicates values ​​between 500 and 2,000 nM *** indicates values ​​between 100 and 500 nM **** indicates a value less than 100 nM - indicates that the value has not been determined Table 2. IL-1β release IC 50 Regarding activity: * indicates values ​​greater than 1,000 nM ** indicates values ​​between 250 and 1,000 nM *** indicates values ​​between 50nM and 250nM **** indicates values ​​less than 50 nM - indicates that the value has not been determined

[0466] 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 listed in the Application Data Sheets, including but not limited to Provisional Application No. 63 / 170,708, filed April 5, 2021, and Provisional Application No. 63 / 185,282, filed May 6, 2021, are incorporated herein by reference in their entirety. The embodiments may be modified, as necessary, to utilize concepts from various patents, applications, and publications to provide further embodiments.

[0467] These and other changes can be made to the embodiments in light of the above Detailed Description. In general, the terms used in the following claims should not be construed to limit the claims to the specific embodiments disclosed in the specification and claims, but should be construed to include all possible embodiments that fall within the full scope of equivalents of the claims. Accordingly, the claims are not limited by this disclosure.

Claims

1. The following structure (I): 【Chemistry 1】 [In the formula, A is C 6 -C 10 Arylene, C 3 -C 10 cycloalkylene, 3- to 10-membered heterocyclylene, or 5- to 6-membered heteroarylene; X is N or CR 4 and; Y is N or CH; Z is N or CH; R 1 is C 1 -C 6 Alkyl, C 1 -C 6 Alkynyl, C 1 -C 6 Hydroxyl alkyl, C 1 -C 6 Carboxyalkyl, C 1 -C 6 Alkoxyalkyl, C 3 -C 10 cycloalkyl, or 3-10 membered heterocyclyl; R 2 is a 3- to 10-membered heterocyclyl, a 3- to 10-membered heterocyclylalkyl, a 3- to 10-membered heterocyclylalkenyl, a 3- to 10-membered heterocyclylcarbonyl, a 3- to 10-membered heterocyclyloxy, or a 5- to 6-membered heteroaryl; R 2 is R 3 If exists, R 2 R bonded to the carbon adjacent to the carbon to which 3 Combined with C 3 -C 8 Forming a cycloalkyl; R 3 When present, each independently represents halogen, cyano, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkoxy, or C 3 -C 8 is cycloalkyl; R 4 , H, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, or C 3 -C 8 is cycloalkyl; and n is 0, 1, 2, 3, or 4. or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof.

2. 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein A is phenylene.

3. 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein A is pyridinylene.

4. In the formula, A is C 3 -C 10 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, which is cycloalkylene or 3- to 10-membered heterocyclylene.

5. In the formula, A is halogen, cyano, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkoxy, or C 3 -C 8 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, substituted with one or more substituents selected from cycloalkyl.

6. 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein A is substituted with one or more halogen substituents.

7. 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein A is unsubstituted.

8. In the formula, X is N or X is CR 4 and R 4 is H or C 1 -C 6 alkyl, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof.

9. In the formula, R 1 But the following structure: 【Chemistry 2】 or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof.

10. The compound has the following structure (Ia): 【Chemistry 3】 2. The compound of claim 1, having the formula:

11. 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein Y is N.

12. 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein Y is CH.

13. The compound has the following structure (Ib): 【Chemistry 4】 2. The compound of claim 1, having the formula:

14. In the formula, R 2 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein:

15. In the formula, R 2 is piperazinyl, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof.

16. In the formula, R 2 has the following structure: 【Chemistry 5】 【change】 or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof.

17. 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein n is 1 or 2.

18. In the formula, R 3 is halogen, cyano, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkoxy, or C 3 -C 8 2. The compound of claim 1 or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein:

19. In the formula, R 3 19. The compound of claim 18, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein is methyl, chloro, fluoro, cyano, difluoromethyl, trifluoromethyl, methoxy, trifluoromethoxy, or cyclopropyl.

20. The compound has the following structure (Ic): 【Chemistry 6】 [In the formula, R 3a is halogen or 3-10 membered heterocyclyl; and n1 is 1, 2, or 3.

2. The compound of claim 1, having the formula:

21. In the formula, R 3a 21. The compound of claim 20, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein is fluoro or piperazinyl.

22. The compound has the following structure (Ic1): 【Chemistry 7】 21. The compound of claim 20, having the formula:

23. 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein Z is N.

24. 2. The compound of claim 1, or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, wherein Z is CH.

25. The structure: 【Chemistry 8】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof.

26. A modulator of the NLRP3 inflammasome, an inhibitor of NEK7, or both, comprising a compound described in any one of claims 1 to 25 or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof.

27. 26. A pharmaceutical composition comprising a compound according to any one of claims 1 to 25 or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof, and a pharma- ceutically acceptable carrier, diluent, or excipient.

28. 26. A medicament for treating an NLRP3 mediated disease, comprising a compound according to any one of claims 1 to 25 or a pharma- ceutically acceptable salt, stereoisomer, or prodrug thereof.

29. 29. The pharmaceutical of claim 28, wherein the disease is selected from autoimmune diseases, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, bacterial and viral infections, allergies, asthma, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, transplantation, sperm motility disorders, red blood cell deficiencies, graft rejection, lung disorders, respiratory diseases, ischemic conditions, and combinations thereof.

30. 29. The pharmaceutical composition of claim 28, wherein the disease is selected from type 2 diabetes, atherosclerosis, Alzheimer's disease, aging, fatty liver, metabolic syndrome, asthma, psoriasis, obesity, acute and chronic tissue damage due to infection, gout, arthritis, macular degeneration, enteritis, hepatitis, peritonitis, silicosis, ultraviolet tanning of the skin, contact hypersensitivity, sepsis, cancer, neurodegenerative diseases, multiple sclerosis, Muckle-Wells syndrome, and combinations thereof.

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