RU2848134C1 - Kinase nek7 inhibitors - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of structure (I):

or a pharmaceutically acceptable salt thereof, where: A is C₆ aryl, C₆ cycloalkyl or 6-membered monocyclic heteroaryl containing from one to two nitrogen heteroatoms, each of which is optionally substituted with one or two R⁶; X is N; Y is NH; R¹ denotes H; R² is , C₃-C₄ cycloalkyl or 4–6 membered heterocyclyl having one heteroatom selected from nitrogen or oxygen, or 6-membered heteroaryl containing one nitrogen heteroatom, where the cycloalkyl or heterocycloalkyl is optionally substituted with one substituent selected from the group consisting of the following: hydroxyl and C₁-C₆ alkyl; R³ denotes H; R⁴ denotes heteroaryl selected from isoxazolyl, 1,2,4-triazolyl, isothiazolyl, 1,2,4-thiadiazolyl and 1,3,4-thiadiazolyl, each of which is optionally substituted with one or more substitutes selected from the group consisting of the following: halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ cyanoalkyl, C₁-C₂ haloalkyl-C₃-C₈ cycloalkyl, C₃ aminylalkylcycloalkyl, C₁-C₂ alkyl-C₃ cycloalkyl and C₁-C₂ alkyl-4-membered heterocyclylalkyl containing a nitrogen or oxygen heteroatom; R⁵ denotes H; and R⁶ in each case independently denotes halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy group, cyano group, C₁-C₆ hydroxyalkyl or C₁-C₆ haloalkyl pharmaceutical compositions containing such compounds, and a method of treating NLRP3-mediated disorders.

EFFECT: kinase nek7 inhibitors are disclosed.

27 cl, 2 tbl, 63 ex

Description

Prerequisites for the creation of the invention

Field of technology to which the invention relates

Embodiments of the present invention generally relate to compounds and methods for their preparation and use as therapeutic or prophylactic agents, such as for the treatment of inflammation.

State of the art

Inflammasomes are multiprotein complexes whose activation plays a key role in innate immunity and inflammation. Four inflammasomes have been described to date: NLRP1, NLRC4, NLRP3, and AIM2. The NLRP3 inflammasome consists of NLRP3, ASC, and caspase-I. Its activation leads to the activation of caspase-1, which promotes the secretion of IL-Iβ and IL-18, cytokines that mediate inflammation in animal models of various autoimmune diseases, myocardial infarction, metabolic syndromes, inflammatory bowel disease, and macrophage activation syndrome.

NEK7 is a member of the NIMA-associated kinase (NEK) family, which acts as NLRP3 binding proteins to regulate their oligomerization and activation. NEK7 is a serine/threonine kinase required for mitotic entry, cell cycle progression, cell division, and mitotic progression. It is expressed in multiple tissues, including the brain, heart, lungs, liver, and spleen. Overexpression of NEK7 induces the production of abnormal cells that are closely associated with tumors such as retinoblastoma, gallbladder cancer, and head and neck carcinoma.

A large number of inhibitors have been widely used to disrupt signaling by effectors including IL-1β or IL-18 without disrupting the inflammatory response. Inhibitors of NLRP3 inflammasome activation, which block the NLRP3-NEK7 interaction, may have therapeutic or preventive activity in various human diseases, such as type 2 diabetes (T2D), atherosclerosis, gout, and neurodegenerative diseases. However, the precise mechanism of NLRP3-NEK7 interaction is not well understood.

Accordingly, it is necessary to develop inhibitors that directly target NEK7 to affect the inflammatory response modulated by the NLRP3 inflammasome in different

pathological diseases such as gout, atherosclerosis, type 2 diabetes, metabolic syndrome, macular degeneration, Alzheimer's disease, multiple sclerosis, and inflammatory bowel disease. Embodiments of the present invention address this issue and provide additional associated benefits.

The essence of the invention

Briefly, embodiments of the present invention relate to compounds, including pharmaceutically acceptable salts, stereoisomers, and prodrugs thereof, that are capable of inhibiting NEK7 and/or modulating the activity of the NLRP3 inflammasome.

One object of the present invention are compounds of structure (I):

pharmaceutically acceptable salts, stereoisomers or prodrugs thereof, wherein all A, X, Y, R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined below.

Another object also is pharmaceutical compositions containing the disclosed compounds and methods of using them for the treatment of inflammation.

Detailed description of the invention

In the following description, certain specific details are provided to clarify various embodiments of the present invention. However, one skilled in the art will appreciate that the present invention may be practiced without these details.

Unless the context otherwise requires, in the present description and claims, the term "include" and its variations such as "includes" and "including" are open ended, inclusive, i.e., mean "including, but not limited to."

In the present description, any concentration range, percentage range, ratio range, or integer range includes any integer value within the range and, where appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise specified. As used herein, the terms "about" and "approximately" mean +20%, +10%, +5%, or +1% of the stated range, value, or structure, unless otherwise specified. It should be understood that the singular terms, as used herein, mean "one or more" of the listed components. The use of an alternative (e.g., "or") means one, both, or any combination of the alternatives.

Reference in this specification to "one 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 invention. Thus, the phrases "in one embodiment" or "in an embodiment" in different parts of the specification do not necessarily refer to the same embodiment. Furthermore, particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

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 invention pertains. As used in this description and claims, singular terms include plural terms unless the context clearly dictates otherwise.

"Amino group" means the radical -NH ₂ .

"Carboxy group" or "carboxyl" means the radical -CO ₂ H.

"Cyano group" means -CN radical.

"Hydroxy" or "hydroxyl" means the radical -OH.

"Nitro group" means the radical -NO ₂ .

"Oxo group" means the substituent =O.

"Thio group" means the -SH substituent.

"Thiooxo group" means the substituent =S.

"Alkyl" means a saturated, straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing from 1 to 12 carbon atoms (C ₁ -C ₁₂ alkyl), from 1 to 8 carbon atoms (C ₁ -C ₈ alkyl) or from 1 to 6 carbon atoms (C ₁ -C ₆ alkyl), or any number within these ranges, such as C ₄ -C ₆ alkyl and the like, and which is attached to the rest of the molecule by a single bond, for example, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (tert-butyl), 3-methylhexyl, 2-methylhexyl and the like. The number of carbon atoms indicated refers to the carbon backbone and the carbon branches, but does not include carbon atoms related to any substituents. Unless otherwise specifically stated in the description, an alkyl group is optionally substituted.

"Alkenyl" means an unsaturated straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, which contains one or more carbon-carbon double bonds, containing from 2 to 12 carbon atoms (C ₂ -C ₁₂ alkenyl), from 2 to 8 carbon atoms (C ₂ -C ₈ alkenyl) or from 2 to 6 carbon atoms (C ₂ -C ₆ alkenyl), or any number within these ranges and which is attached to the rest of the molecule by a single bond, for example, ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like. The number of carbon atoms indicated refers to the carbon backbone and the carbon branches, but does not include carbon atoms belonging to any substituents. Unless otherwise specifically stated in the description, an alkenyl group is optionally substituted.

The term "alkynyl" means an unsaturated linear or branched hydrocarbon radical containing from 2 to 12 carbon atoms (C ₂ -C ₁₂ alkynyl), from 2 to 9 carbon atoms (C ₂ -C ₉ alkynyl) or from 2 to 6 carbon atoms (C ₂ -C ₆ alkynyl), or any number in these ranges and containing at least one carbon-carbon triple bond. Examples of alkynyl groups can be selected from the group consisting of the following: ethynyl, propargyl, but-1-ynyl, but-2-ynyl, etc. The number of carbon atoms indicated refers to the carbon backbone and carbon branches, but does not include carbon atoms belonging to any substituents. Unless otherwise specifically stated in the description, an alkynyl group is optionally substituted.

"Alkoxy" means a radical of the formula ^_OR _a where R _a is an alkyl radical as defined above, containing from 1 to 12 carbon atoms (C ₁ -C ₁₂ alkoxy), from 1 to 8 carbon atoms (C ₁ -C ₈ alkoxy), or from 1 to 6 carbon atoms (C ₁ -C ₆ alkoxy), or any number within these ranges. Unless otherwise specifically stated in the description, an alkoxy group is optionally substituted.

"Aminyl" means a radical of the formula -NR _a R _b , where R _a and R b are both independently H or C ₁ -C ₆ alkyl as defined above. If both R _a and R _b are H, "aminyl" is the same as "amino" as defined above. The C ₁ -C ₆ alkyl moiety of the aminyl group is optionally substituted unless otherwise noted.

"Aminylalkylcycloalkyl" means a radical of the formula -R _a R _b NR _c R _d wherein R _a is cycloalkyl as defined herein, R _b is C ₁ -C ₆ alkyl, R _c is H or C ₁ -C ₆ alkyl, and R _d is C ₁ -C ₆ alkyl as defined above. The cycloalkyl and each C ₁ -C ₆ alkyl portion of the aminylalkylcycloalkyl group are optionally substituted unless otherwise indicated.

"Aromatic ring" means a cyclic planar molecule or molecular moiety (i.e., a radical) with resonant bonds within the ring that have increased stability compared to other bonding arrangements of the same set of atoms. Typically, aromatic rings comprise a set of covalently bonded atoms in the same plane and contain a number of π-electrons (e.g., alternating double and single bonds) that is even but not a multiple of 4 (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 specifically stated in the description, "aromatic ring" includes all radicals that are optionally substituted.

"Aryl" means a carbocyclic ring system radical containing from 6 to 18 carbon atoms, for example from 6 to 10 carbon atoms ( _C6 - _C10 aryl) and at least one carbocyclic aromatic ring. For embodiments of the present invention, the aryl radical is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems. Aryl radicals include, but are not limited to, aryl radicals formed from the following: aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, asi-indacene, sim-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleyadene, pyrene, and triphenylene. Unless otherwise specifically stated in the description, an aryl group is optionally substituted.

"Cyanoalkyl" means an alkyl group containing at least one cyano substituent. The -CN substituent may be located on a primary, secondary, or tertiary carbon atom. Unless otherwise specified, a cyanoalkyl group is optionally substituted.

"Carbocyclic" or "carbocycle" means a ring system where all ring atoms are carbon atoms.

"Cycloalkyl" means a non-aromatic monocyclic or polycyclic carbocyclic radical consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems containing from 3 to 15 ring carbon atoms ( _C3 - _C15 cycloalkyl), from 3 to 10 ring carbon atoms ( _C3 - _C10 cycloalkyl), or from 3 to 8 ring carbon atoms ( _C3 - _C8 cycloalkyl), or any number within these ranges, for example, from 3 to 4 carbon atoms ( _C3 - _C4 cycloalkyl), and which is saturated or partially unsaturated and attached to the rest of the molecule by a single bond. Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Polycyclic radicals include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethylbicyclo[2.2.1]heptanyl, etc. Unless otherwise specifically stated herein, a cycloalkyl group is optionally substituted.

"Alkylcycloalkyl" means a radical group of the formula -R _a R _b where R _a is a cycloalkyl group and R _b is an alkyl group as defined above. Unless otherwise specifically indicated herein, an alkylcycloalkyl group is optionally substituted.

"Fused" means any ring structure described in the present invention that is fused to another ring structure.

"Halogen" means bromine, chlorine, fluorine, or iodine.

"Haloalkyl" means an alkyl radical as defined above that is substituted by one or more halide radicals 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 specifically stated in the description, a haloalkyl group is optionally substituted.

"Halocycloalkyl" means a cycloalkyl radical as defined above that is substituted by one or more halide radicals 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 specifically stated in the description, a halocycloalkyl group is optionally substituted.

"Haloalkylcycloalkyl" means a radical group of the formula -R _a R _b where R _a is a cycloalkyl group and R _b is a haloalkyl group as defined above. Unless otherwise specifically indicated herein, a haloalkylcycloalkyl group is optionally substituted.

"Hydroxyalkyl" means an alkyl radical, as defined above, that is substituted by one or more hydroxyl radicals. The hydroxyalkyl radical is attached to the main chain through an alkyl carbon atom. Unless otherwise specified, a hydroxyalkyl group is optionally substituted.

"Heterocyclyl" means a 3- to 18-membered, such as 3- to 10-membered or 3- to 8-membered, non-aromatic ring radical containing from 1 to 10 ring carbon atoms (e.g., from 2 to 10) and from 1 to 6 ring heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. Unless otherwise specifically stated in the description, a heterocyclyl radical is partially or fully saturated and represents a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused, spirocyclic, and/or bridged ring systems. The nitrogen, oxygen, and sulfur atoms in a heterocyclyl radical are optionally oxidized, and the nitrogen atoms may be optionally quaternized. Examples of such heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, furanonyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, hexahydro-1H-pyrrolizine, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, oxiranyl, piperidinyl, piperazinyl, 4-piperidonyl, azetidinyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxothiomorpholinyl and 1,1-dioxothiomorpholinyl. Unless otherwise specified in the description, the heterocyclyl group is optionally substituted.

"Halogenoheterocyclylalkyl" means a radical group of the formula -R _a R _b where R _a is an alkyl group and R _b is a halogenoheterocyclyl group as defined herein. Unless otherwise specifically indicated herein, a halogenoheterocyclylalkyl group is optionally substituted.

"Heterocyclylalkyl" means a radical group of the formula -R _a R _b where R _a is an alkyl group and R _b is a heterocyclyl group as defined herein. Unless otherwise specifically indicated herein, a heterocyclylalkyl group is optionally substituted.

"Heteroaryl" means a 5- to 18-membered, for example 5- to 6-membered, ring system radical containing from 1 to 13 ring carbon atoms, from 1 to 6 ring heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and at least one aromatic ring. Heteroaryl radicals may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, oxygen and sulfur atoms in the heteroaryl radical are optionally oxidized and the nitrogen atom may 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, indazolyl, 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 otherwise specifically stated in the description, a heteroaryl group is optionally substituted.

Oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl and 1,3,4-thiadiazolyl mean the following structures, respectively:

wherein the oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl and 1,3,4-thiadiazolyl are attached to the remainder of the molecule by a covalent bond to one of the ring carbon atoms of the oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl and 1,3,4-thiadiazolyl.

The term "substituted" as used herein means any of the above groups (e.g., alkyl, alkenyl, alkylene, alkylcarbonyl, alkoxy, alkoxyalkyl, aminylalkyl, aryl, cyanoalkyl, cycloalkyl, haloalkyl, heterocyclyl, heterocyclene, heterocyclylalkyl, heteroaryl, heteroarylalkyl, and/or hydroxyalkyl) wherein at least one hydrogen atom (e.g., 1, 2, 3, or all of the hydrogen atoms) is replaced by a bond to a non-hydrogen substituent. Examples of non-hydrogen substituents include, but are not limited to, amino, carboxyl, cyano, hydroxyl, halogen, nitro, oxo, thiol, thiooxo, alkyl, alkenyl, alkylcarbonyl, alkoxy, aryl, cyanoalkyl, cycloalkyl, haloalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, and/or hydroxyalkyl, each of which may also be optionally substituted with one or more of the above substituents.

In some preferred embodiments, the optional substituents are independently selected from the group consisting of halogen, hydroxyl, cyano, aminyl, _C1 - _C6 alkyl, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C1 - _C6 haloalkyl, _C3 - _C8 cycloalkyl, _C3 - _C8 halocycloalkyl, _C6 - _C10 aryl, 5- or 6-membered heteroaryl, _C1 - _C6 alkoxy, and 3- to 8-membered heterocyclyl.

The term "effective amount" or "therapeutically effective amount" means an amount of a compound described herein that is sufficient to produce the intended effect, including, but not limited to, the treatment of a disease as defined below. A therapeutically effective amount may vary depending on the intended treatment (in vivo) or the subject and disease being treated, such as the subject's body weight or age, the severity of the disease, the route of administration, etc., which can be readily determined by one of ordinary skill in the art. The term also refers to a dose that will result in a specific response in target cells, such as a decrease in platelet aggregation and/or cell migration. The specific dose varies depending on the specific compounds selected, the intended administration regimen, whether administered in combination with other compounds, the time of administration, the tissue into which they are administered, and the physical delivery system in which they are contained.

As used herein, "treatment" or "treating" means an approach to obtaining beneficial or desired results for a disease, disorder, or pathological condition, including, but not limited to, a therapeutic effect and/or a prophylactic effect. A therapeutic benefit means the elimination or alleviation of the existing disorder being treated. Furthermore, a therapeutic benefit is provided by the elimination or alleviation of one or more physiological symptoms associated with the existing disorder, such that an improvement is observed in the subject regardless of the subject's afflictions with the existing disorder. A prophylactic effect includes delaying or eliminating the manifestation of a disease or pathological condition, delaying or eliminating the onset of symptoms of a disease or pathological condition, slowing, stopping, or reversing the progression of a disease or pathological condition, or any combination thereof. In some embodiments, for a prophylactic benefit, the compositions are administered 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 a diagnosis of the disease may not have been made.

The terms "co-administration," "administration in combination with," and their grammatical equivalents, when used herein, include the administration of two or more agents to an animal, including humans, such that both agents and/or their metabolites are present in the subject simultaneously. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in a composition containing both agents.

"Pharmaceutically acceptable salt" includes acid and base addition salts.

"Pharmaceutically acceptable acid addition salt" means those salts that retain the biological effectiveness of the free bases and that are biologically tolerable or otherwise biologically suitable for administration to a subject. See S.M. 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 those that are pharmacologically effective and suitable for contact with the tissues of patients without undesirable toxicity, irritation, or allergic reaction. Pharmaceutically acceptable acid addition salts are formed 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, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, Formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic 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.

"Pharmaceutically acceptable base addition salt" means those salts that retain the biological effectiveness of the free acids and that are biologically tolerable or otherwise biologically suitable for administration to a subject. See S. M. 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 those that are pharmacologically effective and suitable for contact with the tissues of patients without undesirable toxicity, irritation, or allergic reaction. Pharmaceutically acceptable base addition salts are prepared by adding an inorganic base or an 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, aluminum, etc. 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, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, 2-dimethylaminoethanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dichlorhexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benetamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dichlorhexylamine, choline, and caffeine.

In some embodiments, pharmaceutically acceptable salts include quaternary ammonium salts, such as alkyl halides of quaternary amines (e.g., methyl bromide).

The terms "antagonist" and "inhibitor" are used interchangeably and refer to a compound capable of inhibiting the biological function of a target protein by inhibiting the activity or expression of a protein such as the NLRP3 inflammasome or NEK7, or the NLRP3 inflammasome-NEK7 association. Accordingly, the terms "antagonist" and "inhibitors" are defined in the context of a biological effect on a target protein. Although preferred antagonists in the present invention specifically interact with (e.g., bind to) the target, compounds that inhibit the biological activity of a target protein by interacting with other members of the signal transduction pathway for which the target protein is a member are also specifically included in this definition. The preferred biological activity inhibited by the antagonist is related to tumor development, growth, or dissemination.

The term "agonist," as used in the present invention, refers to a compound capable of initiating or enhancing the biological function of a target protein by inhibiting the activity or expression of the protein. Accordingly, the term "agonist" is defined in the context of a biological effect on a target polypeptide. Although preferred agonists in the present invention specifically interact (e.g., bind) with the target, compounds that initiate or enhance the biological activity of a target polypeptide by interacting with other members of the signal transduction pathway of which the target polypeptide is a member are also specifically included in this definition.

"Signal transduction" is the process by which stimulatory or inhibitory signals are transmitted into and within a cell to produce an intracellular response.

The term "selective inhibition" or "selectively inhibit" a biologically active agent means the ability of the agent to preferentially reduce a target signaling activity over non-target signaling activity by directly or indirectly interacting with the target.

"Subject" means an animal, such as a mammal, for example, a human. The methods described in the present invention can be applied in human and veterinary medicine. In some embodiments, the subject is a mammal, and in some embodiments, the subject is a human.

"Mammal" includes humans and domestic animals such as laboratory animals and pets (e.g., cats, dogs, pigs, cattle, sheep, goats, horses, rabbits) and non-domestic animals such as wild animals, etc.

"Prodrug" means a compound that can be converted under physiological conditions or by solvolysis into a biologically active compound as described herein (e.g., compounds of structure (I)). Thus, the term "prodrug" means a precursor of a biologically active compound that is pharmaceutically acceptable. In some instances, a prodrug is inactive when administered to a subject, but is typically converted in vivo into the active compound, for example, by hydrolysis. A prodrug compound often provides advantages of solubility, tissue compatibility, or sustained release in a mammal (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," A.C.S. 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 entireties. The term "prodrug" also includes any covalently linked carriers that release the active compound in vivo when such prodrug is administered to a mammal. Prodrugs of the active compound described in the present invention are typically prepared by modifying functional groups of the active compound in such a way that the modifying moiety can be cleaved by standard procedures or in vivo to yield the parent active compound. Prodrugs include compounds in which a hydroxy, amino, or thio group is linked to any group that, upon administration of the prodrug of the active compound to a mammal, is cleaved to form a free hydroxy, free amino, or free mercapto group, respectively. Examples of prodrugs include, but are not limited to, the acetate, formate, and benzoate of the hydroxy group or the acetamide, formamide, and benzamide of the amino group in the active compound, etc.

The term "in vivo" means an event that occurs in the subject's body.

The embodiments disclosed in the present invention also include all pharmaceutically acceptable compounds of structure (I).

Some embodiments also include in vivo metabolic products of the disclosed compounds. Such products may be formed, for example, by oxidation, reduction, hydrolysis, amidation, esterification, etc., of the administered compound, primarily by enzymatic processes. Accordingly, embodiments include compounds produced by a method comprising administering a compound of the present invention to a mammal for a period of time sufficient to form a product of its metabolism. Such products are typically detected by administering a radioactively labeled compound of the present invention at a detectable dose to an animal, such as a rat, mouse, guinea pig, monkey, or human, allowing time for metabolism to occur and for the products of its metabolism to be excreted from urine, blood, or other biological samples.

"Stable compound" and "stable structure" mean a compound that is sufficiently stable when isolated in suitable purity from a reaction mixture and prepared into an effective therapeutic agent.

Crystallization often yields a solvate of the compounds disclosed herein. As used herein, the term "solvate" refers to an aggregate that comprises one or more compounds provided herein with 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. Thus, the compounds provided herein can exist as a hydrate, including a monohydrate, a dihydrate, a hemihydrate, a sesquihydrate, a trihydrate, a tetrahydrate, and the like, as well as corresponding solvated forms. In some aspects, the compounds provided herein are a true solvate, while in other cases, the compounds provided herein simply retain additional water or are a mixture of water with some additional solvent.

"Optional" or "optionally" means that the event described thereafter may or may not occur and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted aryl" means that the aryl radical may or may not be substituted and that the description includes substituted aryl radicals and aryl radicals that do not contain a substituent.

"Pharmaceutical composition" means a preparation of the compounds of the present invention and a vehicle generally accepted in the art for delivering the compounds of the present invention to mammals, such as humans. Such a vehicle includes all pharmaceutically acceptable carriers, diluents, or excipients.

"Pharmaceutically acceptable carrier, diluent, or excipient" includes, without limitation, any excipient, vehicle, excipient, glidant, sweetener, diluent, preservative, dye/coloring agent, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solubilizer, or emulsifier.

"Stereoisomers" mean compounds consisting of the same atoms linked by the same bonds but having different three-dimensional structures that are not interchangeable. The present invention includes various stereoisomers and mixtures thereof, and includes "enantiomers," which mean two stereoisomers whose molecules are non-superimposable mirror images of each other.

The compounds of the present invention (i.e., the compounds of structure (I)) or pharmaceutically acceptable salts thereof may contain one or more centers of geometric asymmetry and may therefore form stereoisomers, such as enantiomers, diastereoisomers, and other stereoisomeric forms that are defined by absolute stereochemical configurations as (R)- or (S)- or as (D)- or (L)- for amino acids. Thus, embodiments include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers can be prepared using chiral synthons or chiral reagents, or separated by conventional techniques, such as chromatography and fractional crystallization. Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor 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 otherwise indicated, the compounds are understood to include both E and Z geometric isomers. Likewise, all tautomeric forms are included.

Embodiments of the present invention include all types of rotational isomers and conformationally constrained states of the compound provided herein. Also included are atropisomers, which are stereoisomers formed by hindered rotation about a single bond, where the energy difference due to steric strain or other contributions creates a rotational barrier that is large enough to allow the isolation of individual conformers. By way of example, some compounds provided herein may exist as mixtures of atropisomers or as purified or enriched atropisomers.

In some embodiments, compounds of structure (I) are mixtures of enantiomers or diastereoisomers. In other embodiments, compounds of structure (I) are substantially a single enantiomer or diastereoisomer.

A "tautomer" involves the displacement of a proton from one atom of a molecule to another atom of the same molecule. Thus, embodiments include tautomers of the disclosed compounds.

The chemical naming and structure diagram generation protocol used in the present invention is a modified form of the I.U.P.A.C. nomenclature system, using ACD/Name Version 9.07 software and/or ChemDraw Professional Version 17.0.0.206 naming software (CambridgeSoft). In the compound chemical names used in the present invention, the substituent group is typically named before the group to which it is attached. For example, cyclopropylethyl includes an ethyl backbone with a cyclopropyl substituent. Except as described below, all bonds are indicated in the chemical structure diagrams in the present invention except for all bonds of certain carbon atoms, which are assumed to be bonded to a number of hydrogen atoms sufficient to saturate the valence.

Connections

The present invention relates to compounds, including pharmaceutically acceptable salts, stereoisomers and prodrugs thereof, that are capable of inhibiting NEK7 and/or modulating the activity of the NLRP3 inflammasome.

Embodiments of the present invention relate to a compound having the following structure (I):

or a pharmaceutically acceptable salt, stereoisomer or prodrug thereof, wherein:

A is _C6 - _C10 aryl, _C3 - _C10 cycloalkyl, 3- to 10-membered heterocyclyl, or 5- to 6-membered monocyclic heteroaryl, each of which is optionally substituted with one or more ^R6 ;

Y stands for CHOH or NH;

R ¹ is H or C ₁ -C ₆ alkyl;

R ² is C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl or 5- or 6-membered heteroaryl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy and 3- to 8-membered heterocyclyl;

R ³ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl or 5- or 6-membered heteroaryl, each of which is optionally substituted with another substituent selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl and C ₁ -C ₆ alkoxy;

R⁴is a heteroaryl selected from the group consisting of: oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl and 1,3,4-thiadiazolyl, each of which is optionally substituted with one or more substituents selected from the group consisting of: halogen, C₁-C₆alkyl, C₂-WITH₆alkenyl, C₂-WITH₆alkynyl, C₁-WITH₆haloalkyl, C₃-WITH₈cycloalkyl, cyano group, aminyl, C₁-C₆hydroxyalkyl, C₁-C₆cyanoalkyl, 3- to 8-membered heterocyclyl, C₃-WITH₈ haloalkylcycloalkyl, C₃-WITH₈ aminolalkylcycloalkyl, C₃-WITH₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered haloheterocyclylalkyl and C₃-WITH₈halocycloalkyl or combinations thereof;

R ⁵ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl, C ₆ -C ₁₀ aryl, or 5- or 6-membered heteroaryl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, and C ₁ -C ₆ alkoxy; and

R ⁶ in each case independently represents halogen, C ₁ -C ₆ alkyl, cyano, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkyl.

In some embodiments of structure (I):

A is _C6 - _C10 aryl, _C3 - _C10 cycloalkyl, 3- to 10-membered heterocyclyl, or 5- to 6-membered monocyclic heteroaryl, each of which is optionally substituted with one or more ^R6 ;

Y stands for CHOH or NH;

R ¹ is H or C ₁ -C ₆ alkyl;

R ² is C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl or 5- or 6-membered heteroaryl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy and 3- to 8-membered heterocyclyl;

R ³ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl or 5- or 6-membered heteroaryl, each of which is optionally substituted with another substituent selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl and C ₂ -C ₆ alkoxy;

R ⁴ is a heteroaryl selected from the group consisting of: oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl and 1,3,4-thiadiazolyl, each of which is optionally substituted with one or more substituents selected from the group consisting of: halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl and C ₃ -C ₈ halocycloalkyl;

R ⁵ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl, C ₆ -C ₁₀ aryl, or 5- or 6-membered heteroaryl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, and C ₁ -C ₆ alkoxy; and

R ⁶ in each case independently represents halogen, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl.

Some more preferred embodiments relate to a compound having the following structure (I):

or a pharmaceutically acceptable salt, stereoisomer or prodrug thereof, wherein:

A is _C6 - _C10 aryl, _C3 - _C10 cycloalkyl, 3- to 10-membered heterocyclyl, or 5- to 6-membered monocyclic heteroaryl, each of which is optionally substituted with one or more ^R6 ;

X stands for CH or N;

Y stands for CHOH or NH;

R ¹ is H or C ₁ -C ₆ alkyl;

R ² is C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl or 5- or 6-membered heteroaryl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy and 3- to 8-membered heterocyclyl;

R³means H, C₁-WITH₆alkyl, C₂-WITH₆alkenyl, C₂-WITH₆alkynyl, C₃-WITH₈cycloalkyl, 3- to 8-membered heterocyclyl, or 5- or 6-membered heteroaryl, each of which is optionally substituted with another substituent selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C₁-WITH₆alkyl, C₂-WITH₆ alkenyl, C₂-WITH₆alkynyl and C₁-WITH₆alkoxy group;

R ⁴ is a heteroaryl selected from the group consisting of: oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl and 1,3,4-thiadiazolyl, each of which is optionally substituted with one or more substituents selected from the group consisting of: halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, C ₃ -C ₈ aminylalkylcycloalkyl, C ₃ -C ₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered haloheterocyclylalkyl and C ₃ -C ₈ halocycloalkyl;

R ⁵ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl, C ₆ -C ₁₀ aryl, or 5- or 6-membered heteroaryl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, and C ₁ -C ₆ alkoxy; and

R ⁶ in each case independently represents halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, cyano, C ₁ -C ₆ hydroxyalkyl or C ₁ -C ₆ haloalkyl.

In one embodiment, R ¹ is H. In other embodiments, R ¹ is C ₁ -C ₆ alkyl, such as methyl.

One embodiment pertains to compounds of structure (I) wherein R ² is branched C ₄ -C ₆ alkyl, C ₃ -C ₄ cycloalkyl, C ₃ -C ₈ heterocyclyl, or 5- or 6-membered heteroaryl, each of which is optionally substituted with another substituent selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy, and 3- to 8-membered heterocyclyl.

Another embodiment relates to compounds of structure (I), wherein R ² is branched C ₄ -C ₆ alkyl, C ₃ -C ₄ cycloalkyl, or C ₃ -C ₈ heterocyclyl, each of which is optionally substituted with another substituent selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy, and 3- to 8-membered heterocyclyl.

In preferred embodiments, R ² is cyclopropyl or oxetanyl, each of which is optionally substituted with one more substituent selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy, and 3- to 8-membered heterocyclyl. In some embodiments, R ² is cyclopropyl. In other embodiments, R ² is oxetanyl. In some embodiments, R ² is unsubstituted cyclopropyl or oxetanyl.

In preferred embodiments, R ² is cyclopropyl, cyclobutyl, pyrrolidinyl, piperidinyl, or oxetanyl, each of which is optionally substituted with one more substituent selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy, and 3- to 8-membered heterocyclyl. In some embodiments, R ² is cyclopropyl. In other embodiments, R ² is oxetanyl. In some embodiments, R ² is unsubstituted cyclopropyl or oxetanyl. In some embodiments, R ² is N-methyl substituted pyrrolidinyl. In some preferred embodiments, R ² is unsubstituted cyclobutyl.

In other embodiments, R ² is branched C ₄ -C ₆ alkyl optionally substituted with another substituent selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy, and 3- to 8-membered heterocyclyl. For example, in some embodiments, R ² is 2-methylpropyl optionally substituted with hydroxyl.

In more preferred embodiments, R ² has one of the following structures:

In some preferred embodiments, R ² has one of the following structures:

In other embodiments, R ³ is H. In other embodiments, R ³ is C ₁ -C ₆ alkyl, such as methyl.

In any of the preceding embodiments, R ⁴ is oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl or 1,3,4-oxadiazolyl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, C ₃ -C ₈ aminylalkylcycloalkyl, C ₃ -C ₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered haloheterocyclylalkyl, and _C3 - _C8 halocycloalkyl, and combinations thereof. For example, in some embodiments, ^R4 is isoxazolyl optionally substituted with one or more substituents selected from the group consisting of halogen, _C1 - _C6 alkyl, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, _C1 - _C6 haloalkyl, _C3 - _C8 cycloalkyl, and _C3 - _C8 halocycloalkyl. In other preferred embodiments, ^R4 comprises the following substituents: _C1 - _C6 alkyl, _C1 - _C6 haloalkyl, _C3 - _C8 cycloalkyl, or _C3 - _C8 halocycloalkyl.

In some embodiments, R ⁴ is oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, thiazolyl, isothiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,4-triazolyl, or 1,3,4-oxadiazolyl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, _C3 - _C8 aminylalkylcycloalkyl, _C3 - _C8 alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered haloheterocyclylalkyl and _C3 - _C8 halocycloalkyl and combinations thereof.

In some embodiments, R ⁴ is isoxazolyl optionally substituted with one or more substituents selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, C ₃ -C ₈ aminylalkylcycloalkyl, C ₃ -C ₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered haloheterocyclylalkyl, and C ₃ -C ₈ halocycloalkyl and combinations thereof.

In some embodiments, R ⁴ is thiazolyl optionally substituted with one or more substituents selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, C ₃ -C ₈ aminylalkylcycloalkyl, C ₃ -C ₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered haloheterocyclylalkyl, and C ₃ -C ₈ halocycloalkyl and combinations thereof.

In some embodiments, R ⁴ is isothiazolyl optionally substituted with one or more substituents selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, C ₃ -C ₈ aminylalkylcycloalkyl, C ₃ -C ₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered haloheterocyclylalkyl, and C ₃ -C ₈ halocycloalkyl and combinations thereof.

In some embodiments, R ⁴ is 1,2,4-thiadiazolyl optionally substituted with one or more substituents selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, C ₃ -C ₈ aminylalkylcycloalkyl, C ₃ -C ₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered halogenoheterocyclylalkyl and _C3 - _C8 halogenocycloalkyl and combinations thereof.

In some embodiments, R ⁴ is 1,3,4-thiadiazolyl optionally substituted with one or more substituents selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, C ₃ -C ₈ aminylalkylcycloalkyl, C ₃ -C ₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered halogenoheterocyclylalkyl and _C3 - _C8 halogenocycloalkyl and combinations thereof.

In some embodiments, R ⁴ is 1,2,4-triazolyl optionally substituted with one or more substituents selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, C ₃ -C ₈ aminylalkylcycloalkyl, C ₃ -C ₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered halogenoheterocyclylalkyl and _C3 - _C8 halogenocycloalkyl and combinations thereof.

In some embodiments, R ⁴ is 1,3,4-oxadiazolyl optionally substituted with one or more substituents selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, 3- to 8-membered heterocyclyl, and C ₃ -C ₈ halocycloalkyl, or combinations thereof.

In some embodiments, R ⁴ comprises the following substituents: C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, C ₃ -C ₈ aminylalkylcycloalkyl, C ₃ -C ₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered haloheterocyclylalkyl and C ₃ -C ₈ halocycloalkyl, and combinations thereof.

In various embodiments, R ⁴ has one of the following structures:

In various other embodiments, R ⁴ has one of the following structures:

In various other embodiments, R ⁴ has one of the following structures:

In some preferred embodiments, R ² is C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl, or 5- or 6-membered heteroaryl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy, and 3- to 8-membered heterocyclyl; and

R ⁴ has one of the following structures:

In some preferred embodiments, R ² is C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl, or 5- or 6-membered heteroaryl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy, and 3- to 8-membered heterocyclyl; and

R ⁴ has one of the following structures:

In more preferred embodiments, R ² is C ₁ -C ₆ alkyl substituted with hydroxyl or C ₁ -C ₆ alkoxy. In some embodiments, R ² has one of the following structures:

In other embodiments, R ⁵ is H. In other embodiments, R ⁵ is C ₁ -C ₆ alkyl, such as methyl.

In some embodiments, Υ is C(H) (OH). In other embodiments, Υ is NH.

In various embodiments, A is _C6 - _C10 aryl, _C3 - _C10 cycloalkyl, or 5- to 6-membered monocyclic heteroaryl, each of which is optionally substituted with one or more ^R6 . It should be understood that A is a divalent radical.

In some embodiments, A is a divalent optionally substituted C _6-10 aryl. In some embodiments, A is a divalent optionally substituted 3- to 8-membered saturated or partially unsaturated carbocyclic ring. In some embodiments, A is a divalent optionally substituted 3- to 10-membered heterocyclic ring containing 1-4 heteroatoms independently selected from the group consisting of nitrogen, oxygen, or sulfur. In some embodiments, A is a divalent optionally substituted 5- to 6-membered monocyclic heteroaryl ring containing 1-4 heteroatoms independently selected from the group consisting of nitrogen, oxygen, or sulfur.

In some embodiments, A is a divalent group selected from the following: phenyl, pyridinyl, cyclohexyl, and cyclohexenyl; each of which is optionally substituted. In other embodiments, A is phenyl. In other embodiments, A is saturated or unsaturated cyclohexyl. In more preferred embodiments, A is pyridinyl.

In some embodiments, A is pyrimidinyl, which is optionally substituted.

In any of the preceding embodiments, A is unsubstituted. In other embodiments, A is substituted with one or more R ⁶ . For example, in some embodiments, R ⁶ is halogen. In some embodiments, R ⁶ is chlorine or fluorine. In other embodiments, R ⁶ is fluorine.

In some embodiments, R ⁶ is C ₁ -C ₆ hydroxyalkyl. In some embodiments, the C ₁ -C ₆ hydroxyalkyl is -CH ₂ CH ₂ OH. In other embodiments, R ⁶ is cyano. In some embodiments, R ⁶ is C ₁ -C ₆ alkoxy. In more preferred embodiments, the C ₁ -C ₆ alkoxy is methoxy.

In some embodiments, A is a divalent group selected from the following: phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctanyl, [4.3.0]bicyclononanyl, [4.4.0]bicyclodecanyl, [2.2.2]bicyclooctanyl, fluorenyl, indanyl, tetrahydronaphthyl, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, dithiazinyl, tetrahydrofuranyl, furanyl, furazanil, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3-indolyl, isoindolinyl, isoindolenyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl; 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinil, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl and xanthenyl; each of which is optionally substituted.

In preferred embodiments, A has one of the following structures:

In some preferred embodiments, A has one of the following structures:

In some preferred embodiments, A has one of the following structures:

In some embodiments, the compound has the following structure (IA):

or a pharmaceutically acceptable salt, stereoisomer or prodrug thereof, wherein:

R ^2a is C ₁ -C ₆ alkyl or C ₃ -C ₈ cycloalkyl, optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy and 3- to 8-membered heterocyclyl;

R ^4a is isoxazolyl optionally substituted with one or more substituents selected from the group consisting of C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl or C ₃ -C ₈ haloalkylcycloalkyl.

In more preferred embodiments, R ^2a is branched C ₁ -C ₆ alkyl substituted with hydroxyl. In some embodiments, R ^2a is C ₃ -C ₈ cycloalkyl. In more preferred embodiments, R ^2a has one of the following structures:

In some embodiments, R ^4a is isoxazolyl substituted with C ₃ -C ₈ haloalkylcycloalkyl. In some embodiments, R ^4a is C ₃ -C ₈ fluoroalkylcycloalkyl. In even more preferred embodiments, R ^4a is fluoroalkylcyclopropyl or fluoroalkylcyclobutyl. In more preferred embodiments, R ^4a has one of the following structures:

In some embodiments, X is CH. In some more preferred embodiments, the compound has the following structure (IB):

or a pharmaceutically acceptable salt, stereoisomer or prodrug thereof, wherein:

A is _C6 - _C10 aryl, _C3 - _C10 cycloalkyl, 3- to 10-membered heterocyclyl, or 5- to 6-membered monocyclic heteroaryl, each of which is optionally substituted with one or more ^R6 ;

X stands for CH or N;

Y stands for CHOH or NH;

R ¹ is H or C ₁ -C ₆ alkyl;

R ² is C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl or 5- or 6-membered heteroaryl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy and 3- to 8-membered heterocyclyl;

R ³ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl or 5- or 6-membered heteroaryl, each of which is optionally substituted with another substituent selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl and C ₁ -C ₆ alkoxy;

R ⁴ is a heteroaryl selected from the group consisting of: oxazolyl, isoxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl and 1,3,4-thiadiazolyl, each of which is optionally substituted with one or more substituents selected from the group consisting of: halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl, cyano, aminyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, 3- to 8-membered heterocyclyl, C ₃ -C ₈ haloalkylcycloalkyl, C ₃ -C ₈ aminylalkylcycloalkyl, C ₃ -C ₈ alkylcycloalkyl, 3- to 8-membered heterocyclylalkyl, 3- to 8-membered alkylheterocyclylcycloalkyl, 3- to 8-membered haloheterocyclylalkyl and C ₃ -C ₈ halocycloalkyl;

R ⁵ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl, C ₆ -C ₁₀ aryl, or 5- or 6-membered heteroaryl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, aminyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, and C ₁ -C ₆ alkoxy; and

R ⁶ in each case independently represents halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, cyano, C ₁ -C ₆ hydroxyalkyl or C ₁ -C ₆ haloalkyl.

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

In a preferred embodiment, the compound of structure (I) is an inhibitor of NEK7 in a patient or in a biological sample.

In various other embodiments, the compound has one of the structures shown below in Table 1, or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof. The compounds in Table 1 were prepared as described in the Examples or by methods known in the art and analyzed using mass spectrometry and/or ^1H NMR.

It should be understood that in the present description, combinations of substituents and/or variables in the given formulas are permissible only if they lead to stable compounds.

In a further embodiment, various compounds of the present invention that exist in free base or acid form can be converted into their pharmaceutically acceptable salts by treatment with a suitable inorganic or organic base or acid using methods known to those skilled in the art. Salts of the compounds of the present invention can be converted into their free bases or acids using standard methods.

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

The following general reaction schemes illustrate examples of compounds of the present invention, structure (I):

or a pharmaceutically acceptable salt, stereoisomer or prodrug thereof, wherein each A, X, Y, R ¹ , R ² , R ³ , R ⁴ and R ⁵ is as defined below.

General reaction scheme 1

The following general reaction scheme, where X ¹ and X ² are independently halogens and X, R ¹ , R ² , R ³ and A have the meanings described herein, illustrates exemplary procedures for preparing the intermediate amine D:

As shown in General Reaction Scheme 1, alkylation of pyrimidine/pyridinepyrrole (i.e., intermediate A) with a cycloalkyl boronate or a suitable electrophile in the presence of a base affords intermediate B. This precursor is treated with ammonium hydroxide to afford the pyrrolopyrimidine/pyridin-4-amine derivative of intermediate C. The resulting intermediate C can be subjected to palladium-catalyzed arylation to afford intermediate D.

General reaction scheme 2

The following general reaction scheme illustrates examples of methods for preparing the intermediate carbamate E:

As shown in General Reaction Scheme 2, intermediate E can be prepared in the presence of base by the reaction of phenyl chloroformates and the indicated heteroarylamine (an amino-substituted analog of R ⁴ ). General Reaction Scheme 2 depicts the preparation of compounds in which R ⁵ is H; however, compounds in which R ⁵ is not H can be prepared by analogous procedures by introducing R ⁵ after the preparation of intermediate E or by using an appropriately substituted heteroarylamine.

General reaction scheme 3

The following general reaction scheme illustrates examples of methods for preparing compounds of structure (I):

Intermediate D and intermediate E are treated with a base (e.g., trimethylamine, DIPEA, DMAP, etc.) in THF to give compounds of structure (I).

General reaction scheme 4

The following general reaction scheme illustrates examples of compounds of structure (I):

Intermediate D is reacted with the said phenyl chloroformate under suitable conditions to give intermediate E. Intermediate E is then coupled with an amine using a suitable base (e.g., trimethylamine, DIPEA, DMAP, etc.) in THF to give compounds of structure (I).

Any of the above reaction schemes can be modified at any stage to add and/or modify a substituent, and the substituent can be added or modified accordingly at any stage of the overall synthesis of the desired compounds.

Those skilled in the art will appreciate that in preparing the compounds described in the present invention, functional groups of the intermediates may need to be protected with suitable protecting groups. Such functional groups include, but are not limited to, hydroxy, amino, mercapto, and carboxy. Suitable protecting groups for the hydroxy group include trialkylsilyl or diarylalkylsilyl (e.g., t-butyldimethylsilyl, t-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for the amino, amidino, and guanidino groups include t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for the mercapto group include -C(O)-R" (where R" is alkyl, aryl, or arylalkyl), p-methoxybenzyl, trityl, and the like. Suitable protecting groups for the carboxy group include alkyl, aryl, or arylalkyl ester groups. Protecting groups are optionally introduced or removed using standard techniques known to those skilled in the art and described herein. The use of protecting groups is described in detail in Green, T.W. and P.G.M. Wutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley. As will be appreciated by those skilled in the art, the protecting group can also be a polymeric resin, such as Wang resin, Rink resin, or 2-chlorotrityl chloride resin.

Those skilled in the art will also appreciate that, although such protected derivatives of the compounds of the present invention may not themselves possess pharmacological activity, they can be administered to a mammal and then metabolized in the body to form the compounds of the present invention, which are pharmacologically active. Therefore, such derivatives can be described as "prodrugs." Prodrugs of the compounds of the present invention are included within the scope of embodiments of the present invention.

Pharmaceutical compositions

Other embodiments relate to pharmaceutical compositions. The pharmaceutical composition comprises one or more of the above compounds and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is prepared for oral administration. In other embodiments, the pharmaceutical composition is prepared for injection. In other embodiments, the pharmaceutical compositions comprise a compound disclosed herein and an additional therapeutic agent (e.g., an anticancer agent). Non-limiting examples of such therapeutic agents are described below in this disclosure.

Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ocular, pulmonary, mucosal, transdermal, vaginal, aural, nasal, and topical administration. Additionally, by way of example only, parenteral delivery includes intramuscular, subcutaneous, intravenous, intramedullary, intrathecal, direct intraventricular, intraperitoneal, intralymphatic, and intranasal injections.

In some embodiments, a compound described herein is administered locally rather than systemically, such as by injecting the compound directly into an organ, often as a depot or sustained-release formulation. In preferred embodiments, long-acting formulations are administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Furthermore, in other embodiments, the compound is delivered using a targeted drug delivery system, such as a coated liposome and an organ-specific antibody. In such embodiments, the liposomes are targeted to the organ and are selectively taken up by the organ. In other embodiments, a compound described herein is used in the form of a quick-release formulation, an extended-release formulation, or an intermediate-release formulation. In other embodiments, a compound described herein is administered locally.

In the treatment methods according to embodiments of the present invention, an effective amount of at least one compound of structure (I) is administered to a subject suffering from or diagnosed as having such a disease, disorder, or pathological condition. Effective amounts or doses can be determined by methods such as modeling, dose escalation studies, or clinical studies, for example, the regimen or route of administration or delivery of the drug, the pharmacokinetics of the drug, the severity and course of the disease, disorder, or pathological condition, the presence of previous or current treatment in the subject, the subject's health status and response to drugs, and the discretion of the treating physician.

The compounds provided in the present invention are effective over a wide dosage range. For example, for the treatment of adults, doses of 10 to 5000 mg, 100 to 5000 mg, 1000 to 4000 mg per day, and 1000 to 3000 mg per day are examples of doses used in some embodiments. The exact dosage depends on the route of administration, the form in which the compound is administered, the subject being treated, the body weight of the subject being treated, and the preference and experience of the treating physician.

In some embodiments, the compounds of the present invention are administered in a single dose. Typically, such administration is by injection, such as intravenous injection, for rapid delivery. However, other routes are also used if suitable. A single dose of a compound of the present invention can also be used to treat an acute medical condition.

In some embodiments, the compounds of the present invention are administered in multiple doses. In some embodiments, administration is about one, two, three, four, five times, 6 times, or more than 6 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 compounds of the present invention and another agent (e.g., an anticancer agent) are administered together from about once a day to about 6 times per day. In another embodiment, administration of the compounds of the present invention and the agent is carried out for less than about 7 days. In another embodiment, administration is carried out for more than about 6, 10, 14, 28 days, two months, 6 months, or one year. In some cases, continuous administration is prescribed and carried out for as long as necessary.

The compounds of the present invention can be administered for as long as necessary. In some embodiments, the compounds of the present invention are administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, the compounds of the present invention are administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, the compounds of the present invention are administered chronically on a continuous basis, for example, to treat chronic effects.

In some embodiments, the compounds of the present invention are administered in individual dosage forms. It is known in the art that, due to differences in compound pharmacokinetics between subjects, individualization of the administration regimen is necessary for optimal treatment.

In some embodiments, the compounds described herein are prepared as pharmaceutical compositions. In preferred embodiments, the pharmaceutical compositions are prepared in a conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliaries that facilitate processing of the disclosed compounds into preparations that can be used pharmaceutically. The appropriate formulation depends on the chosen route of administration. Any pharmaceutically acceptable techniques, carriers, and excipients are used as appropriate for preparing the pharmaceutical compositions described herein: Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999).

The present invention relates to pharmaceutical compositions comprising one or more compounds of structure (I) and a pharmaceutically acceptable carrier.

The present invention relates to pharmaceutical compositions comprising one or more compounds selected from compounds of structure (I) and a pharmaceutically acceptable diluent(s), excipient(s), and carrier(s). In some embodiments, the compounds described are administered as pharmaceutical compositions in which one or more compounds selected from compounds of structure (I) are mixed with other active ingredients, as in combination therapy. The present invention includes all combinations of active compounds specified in the section on combination therapy below and throughout the present invention. In preferred embodiments, the pharmaceutical compositions comprise one or more compounds of structure (I).

In one embodiment, the pharmaceutical compositions of the compounds of structure (I) are modulators of the NLRP3 inflammasome.

In a preferred embodiment, pharmaceutical compositions of compounds of structure (I) inhibit NEK7 when administered to a patient or a biological sample.

A pharmaceutical composition, as used in the present invention, means a mixture of one or more compounds selected from among the compounds of structure (I) with other chemical components such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents and/or excipients. In some embodiments, the pharmaceutical composition facilitates the administration of the compound to the body. In some embodiments, therapeutically effective amounts of one or more compounds selected from among the compounds of structure (I) provided in the present invention are administered in a pharmaceutical composition to a mammal that has the disease, disorder or pathological condition being treated. In preferred embodiments, the mammal is a human. In some embodiments, therapeutically effective amounts vary depending on the severity of the disease, the age and relative health of the subject, the activity of the compound used and other factors. The compounds described in the present invention are used alone or in combination with one or more therapeutic agents as components of mixtures.

In one embodiment, one or more compounds selected from among the compounds of structure (I) are prepared in aqueous solutions. In preferred embodiments, the aqueous solution is selected from among the following, by way of example only, a physiologically compatible buffer, such as Hank's solution, Ringer's solution, or physiological buffered saline. In other embodiments, one or more compounds selected from among the compounds of structure (I) are prepared for transmucosal administration. In preferred embodiments, preparations for transmucosal administration include penetrating agents that are suitable for permeation through the barrier. In other embodiments, in which the compounds described in the present invention are prepared for other parenteral injections, suitable preparations include aqueous or non-aqueous solutions. In preferred embodiments, such solutions include physiologically compatible buffers and/or inert excipients.

In another embodiment, the compounds described herein are prepared for oral administration. The compounds described herein are prepared by combining the active compounds with, for example, pharmaceutically acceptable carriers or excipients. In various embodiments, the compounds described herein are prepared in oral dosage forms, which include, by way of example only, tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, dispersions, suspensions, and the like.

In some embodiments, pharmaceutical preparations for oral use are obtained by mixing one or more solid inert excipients with one or more compounds described in the present invention, optionally milling the resulting mixture and processing the mixture into granules after adding suitable excipients, if desired, to obtain tablets or dragee cores. Suitable inert excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulose preparations such as: for example, corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, microcrystalline cellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose; or others, such as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. In preferred embodiments, disintegrating agents are optionally added. Disintegrating agents include, by way of example only, cross-linked sodium croscarmellose, polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

In one embodiment, dosage forms such as dragee cores and tablets are prepared with one or more suitable coatings. In preferred embodiments, concentrated sugar solutions are used as coatings for the dosage form. The sugar solutions optionally contain additional components, such as, by way of example only, gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Colorants and/or pigments are also optionally added to the coatings for identification. Furthermore, colorants and/or pigments are optionally used to characterize different dose combinations of the active compound.

In some embodiments, therapeutically effective amounts of at least one compound described herein are prepared in other oral dosage forms. Oral dosage forms include collapsible capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerin or sorbitol. In preferred embodiments, the collapsible capsules contain the active ingredients in admixture with one or more fillers. Fillers include, by way of example only, lactose, binders such as starches, and/or lubricants such as 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 of the following: fixed oil, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers are optionally added.

In other embodiments, the compounds described herein are prepared for parenteral injection, including preparations suitable for bolus injection or continuous infusion. In preferred embodiments, preparations for injection are prepared in unit dosage forms (e.g., in ampoules) or in multi-dose containers. Preservatives are optionally added to preparations for injection. In other embodiments, pharmaceutical compositions are prepared in a form suitable for parenteral injection as sterile suspensions, solutions, or emulsions in oily or aqueous vehicles. Preparations for parenteral injection optionally contain formulation auxiliary agents such as suspending, stabilizing, and/or dispersing agents. In preferred embodiments, pharmaceutical preparations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. In additional embodiments, suspensions of one or more compounds selected from among the compounds of structure (I), if appropriate, are prepared as oily injection suspensions. Suitable lipophilic solvents or diluents for use in the pharmaceutical compositions described herein include, by way of example only, fixed oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. In some preferred embodiments, aqueous injection suspensions contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. The suspension optionally contains suitable stabilizers or agents that increase the solubility of the compounds to enable the preparation of highly concentrated solutions. Alternatively, in other embodiments, the active ingredient is in powder form for reconstitution with a suitable diluent, such as sterile, pyrogen-free water, before use.

The pharmaceutical compositions comprise at least one pharmaceutically acceptable carrier, diluent, or excipient and one or more compounds selected from the compounds of structure (I) described in the present invention as the active ingredient. The active ingredient is in the form of a free acid or free base, or in the form of a pharmaceutically acceptable salt. In addition, the methods and pharmaceutical compositions described in the present invention include the use of N-oxides, crystalline forms (also known as polymorphs), as well as active metabolites of these compounds possessing the same type of activity. All tautomers of the compounds described in the present invention are included within the scope of the compounds provided in the present invention. In addition, the compounds described in the present invention include unsolvated forms, as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. Solvated forms of the compounds provided in the present invention are also considered to be disclosed in the present invention. In addition, the pharmaceutical compositions optionally include other medicinal or pharmaceutical agents, carriers, auxiliary substances such as preservatives, stabilizing, wetting or emulsifying agents, dissolution promoters, salts for regulating osmotic pressure, buffers and/or other therapeutically valuable substances.

Methods for preparing compositions containing the compounds described in the present invention include preparing the compounds together with one or more inert, pharmaceutically acceptable excipients or carriers to form a solid, semi-liquid, or liquid composition. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, wafers, and suppositories. Liquid compositions include solutions in which the compound is dissolved, emulsions containing the compound, or a solution containing liposomes, micelles, or nanoparticles containing a compound disclosed in the present invention. Semi-liquid compositions include, but are not limited to, gels, suspensions, and creams. The form of the pharmaceutical compositions described in the present invention includes liquid solutions or suspensions, solid forms suitable for preparing a solution or suspension in liquid form before use, or emulsions. These compositions also optionally contain small amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, etc.

In some embodiments, pharmaceutical compositions containing one or more compounds selected from compounds of structure (I) are illustratively in liquid form, wherein the agents are contained in solution, suspension, or both. Typically, if the composition is administered as a suspension, the first portion of the agent is in solution and the second portion of the agent is in particulate form, suspended in a liquid matrix. In some embodiments, the liquid composition includes a gel formulation. In other embodiments, the liquid composition is aqueous.

In some embodiments, aqueous suspensions contain one or more polymers as suspending agents. Polymers include water-soluble polymers, such as cellulose polymers, for example, hydroxypropyl methylcellulose, and water-insoluble polymers, such as cross-linked carboxyl-containing polymers. Some pharmaceutical compositions described in the present invention contain a mucoadhesive polymer selected, for example, from the following: carboxymethylcellulose, carbomer (an acrylic acid polymer), poly(methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid/butyl acrylate copolymer, sodium alginate, and dextran.

Pharmaceutical compositions also optionally include solubilizing agents to improve the solubility of one or more compounds selected from compounds of structure (I). The term "solubilizing agent" generally includes agents that result in the formation of a micellar solution or a true solution of the agent. Certain suitable nonionic surfactants, such as polysorbate 80, are useful as solubilizing agents, as are ophthalmologically acceptable glycols, polyglycols such as polyethylene glycol 400, and glycol esters.

In addition, the pharmaceutical compositions optionally include one or more pH adjusting agents or buffering reagents, including acids such as acetic, boric, citric, lactic, phosphoric, and hydrochloric acids; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and tris-hydroxymethylaminomethane; and buffers such as citrate/dextrose, sodium bicarbonate, and ammonium chloride. Such acids, bases, and buffers are included in an amount necessary to maintain the pH of the composition within an acceptable range.

The compositions also optionally include one or more salts in an amount necessary to adjust the osmolality of the composition to an acceptable range. Such salts include those containing sodium, potassium, or ammonium cations and anions such as chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate, or bisulfite; suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.

Other pharmaceutical compositions optionally include one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as merphen and thiomersal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

The compositions may include one or more surfactants to enhance physical stability or for other purposes. Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, such as polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkyl ethers and alkyl phenyl ethers, such as octoxynol 10 and octoxynol 40.

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

In some embodiments, aqueous suspension compositions are packaged in single-dose, non-resealable containers. Alternatively, multi-dose, resealable containers are used, in which case a preservative is typically included in the composition.

Alternative embodiments utilize other delivery systems for hydrophobic pharmaceutical compounds. Liposomes and emulsions are examples of diluents or carriers useful in the present invention. Some embodiments also utilize organic solvents, such as N-methylpyrrolidone. In additional embodiments, the compounds described herein are delivered using an extended-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. The present invention utilizes various extended-release materials. In some embodiments, extended-release capsules release compounds over a period of several weeks to over 100 days. Depending on the chemical nature and biological stability of the therapeutic agent, additional strategies for protein stabilization are utilized.

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

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

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

In some embodiments, the amount of one or more compounds selected from the compounds of structure (I) contained in the pharmaceutical compositions of the present invention is less than or equal to 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, 0.3 g, 0.25 g, 0.2 g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03 g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g, 0.004 g, 0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g, 0.0005 g, 0.0004 g, 0.0003 g, 0.0002 g or 0.0001 g.

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

Packaging materials used for packaging the pharmaceutical compositions described in the present invention 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 suitable for the selected formulation and mode of administration and treatment. For example, the container(s) include one or more compounds described in the present invention, optionally in a composition or in combination with another agent disclosed in the present invention. The container(s) optionally include a sterile entry port (e.g., the container is a sterile bag of intravenous solution or a vial with a stopper pierced by a hypodermic injection needle). Such kits optionally contain a compound with an identifying description or label or instructions for its use in the methods described in the present invention.

For example, a kit typically includes one or more additional containers, each with one or more different materials (such as reagents, optionally in concentrated form, and/or devices) desirable from a commercial and user standpoint for using the compound described in the present invention. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, filters, needles, syringes; a carrier, packaging, container, vial, and/or tube with labels indicating the contents and/or instructions for use, and package inserts with instructions for use. A set of instructions for use is typically also included. The label is optionally located on or together with the container. For example, the label is located on the container, wherein the letters, numbers, or other icons forming the label are attached, molded, or etched on the container itself; the label is associated with the container when it is in the box or carrier so that it also holds the container, for example, as a package insert. In addition, the label is used to indicate that the contents are to be used for a specific therapeutic application. Furthermore, the label provides directions for using the contents, for example, in the methods described herein. In some embodiments, the pharmaceutical compositions are contained in a package or dispenser device that contains one or more unit dosage forms containing a compound of the present invention. The package, for example, comprises a metal or plastic foil, such as a blister pack. Or the package or dispenser contains instructions for administration. Or the package or dispenser contains a direction associated with the container in a form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, and this direction contains approval by the agency for use of a form of the drug for administration to humans or animals. Such a direction, for example, is labeling approved by the U.S. Food and Drug Administration for the prescription of drugs, or an approved package insert. In some embodiments, compositions comprising a compound of the present invention formulated in a compatible pharmaceutical carrier are placed in a suitable container and labeled for the treatment of a specified pathological condition.

Methods

Embodiments of the present invention are useful as modulators of the NLRP3 inflammasome by inhibiting NEK7 in the host species. Therefore, compounds of structure (I) are also useful for the treatment of pathological conditions mediated by effector signaling molecules such as Il-β and IL-18.

The host or patient may be any mammalian species, such as primates, particularly humans; rodents, including mice, rats, and hamsters; rabbits; horses, cows, dogs, cats, etc. Animal models are of interest to experimenters, forming a model for the treatment of human diseases.

In one embodiment, the present invention is useful as an inhibitor of the NLRP3 inflammasome activation mechanism. Therefore, compounds of structure (I) are also useful for the treatment of pathological conditions associated with this activation in host species.

In another embodiment, compounds of structure (I) are useful as inhibitors of the NLRP3 (protein)-NEK7 (protein) interaction. Therefore, the compounds are also useful for the treatment of pathological conditions associated with the NLRP3-NEK7 association in host species.

In some embodiments, the compounds of structure (I) are useful for treating human pathological conditions mediated by effectors selected from the group consisting of IL-β, IL-18, and caspase-1.

Embodiments of the present invention also relate to the use of compounds of structure (I) and/or their physiologically acceptable salts for the prophylactic or therapeutic treatment and/or monitoring of diseases that are caused, mediated and/or modulated by the activity of the NLRP3 inflammasome. Furthermore, embodiments of the present invention relate to the use of compounds of structure (I) and/or their physiologically acceptable salts for the preparation of a medicament for the prophylactic or therapeutic treatment and/or monitoring of diseases that are caused, mediated and/or modulated by the activity of the NLRP3 inflammasome. In some embodiments, the present invention relates to the use of compounds of structure I and/or their physiologically acceptable salts for the preparation of a medicament for the prophylactic or therapeutic treatment of an NLRP3-mediated disorder.

In another embodiment, the present invention relates to a method for treating inflammatory diseases or pathological conditions mediated by the NLRP3 inflammasome by administering to a patient in need thereof a therapeutically effective amount of a compound of structure (I).

In some embodiments, diseases that can be treated with a compound of structure (I) include type II diabetes, atherosclerosis, Alzheimer's disease, aging, fatty liver disease, metabolic syndrome, asthma, psoriasis, obesity, acute and chronic tissue damage caused by infection, gout, arthritis, enteritis, hepatitis, peritonitis, silicosis, UV-induced sunburn, contact hypersensitivity, sepsis, cancer, neurodegenerative disease, multiple sclerosis, and Maki-Wells syndrome.

In some other embodiments, compounds of structure (I) are useful in methods of treating disorders or diseases selected from the group consisting of the following: autoimmune, inflammatory disorders, cardiovascular diseases, neurodegenerative disorders, bacterial and viral infections, allergies, asthma, pancreatitis, multiple organ failure, kidney diseases, platelet aggregation, transplantation, sperm motility, red blood cell deficiency, transplant rejection, lung lesions, respiratory diseases, ischemic conditions, and cancer. In some more preferred embodiments, compounds of structure (I) are useful in methods of treating myelodysplastic syndrome (MDS).

In some embodiments, the NEK7-associated disorder treated with the compound of structure (I) is selected from the group consisting of: rheumatoid arthritis, psoriatic arthritis, osteoarthritis, systemic lupus erythematosus, lupus nephritis, ankylosing spondylitis, osteoporosis, systemic sclerosis, multiple sclerosis, psoriasis, type I diabetes, type II diabetes, inflammatory bowel disease (Crohn's disease and ulcerative colitis), hyperimmunoglobulinemia D and intermittent fever syndrome, cryopyrin-associated periodic syndromes, Schnitzler syndrome, systemic juvenile idiopathic arthritis, adult-onset Still's disease, gout, pseudogout, SAPHO syndrome, Castleman disease, sepsis, stroke, atherosclerosis, celiac disease, DIRA (IL-1 receptor antagonist deficiency), Alzheimer's disease, Parkinson's disease, and cancer.

Also included in the present invention 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, non-specific and COX-2 specific cyclooxygenase inhibitors, gold compounds, corticosteroids, methotrexate, tumor necrosis factor (TNF) antagonists, immunosuppressants, and methotrexate. Examples of NSAIDs include, but are not limited to, ibuprofen, flurbiprofen, naproxen and naproxen sodium, diclofenac, combinations of diclofenac sodium and misoprostol, sulindac, oxaprozin, diflunisal, piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen, nabumetone sodium, sulfasalazine, tolmetin sodium, and hydroxychloroquine.

Examples of NSAIDs also include COX-2 specific inhibitors such as celecoxib, valdecoxib, lumiracoxib and/or etoricoxib.

In some embodiments, the anti-inflammatory agent is a salicylate. Salicylates include, but are not limited to, acetylsalicylic acid or aspirin, sodium salicylate, and choline and magnesium salicylates.

A corticosteroid can also be an anti-inflammatory drug. For example, corticosteroids can include cortisone, dexamethasone, methylprednisolone, prednisolone, prednisolone sodium phosphate, or prednisone.

In further embodiments, the anti-inflammatory agent is a gold compound, such as gold sodium thiomalate or auranofin.

The present invention also includes embodiments in which the anti-inflammatory agent is a metabolic inhibitor, such as a dihydrofolate reductase inhibitor, such as methotrexate, or a dihydroorotate dehydrogenase inhibitor, such as leflunomide.

Therapeutic agents may include anti-pain and anti-inflammatory agents such as histamine and histamine antagonists, bradykinin and bradykinin antagonists, 5-hydroxytryptamine (serotonin), lipid substances that are formed during the biological transformations of the products of selective hydrolysis of membrane phospholipids, eicosanoids, prostaglandins, thromboxanes, leukotrienes, aspirin, non-steroidal anti-inflammatory drugs, analgesic-antipyretic agents, agents that inhibit the synthesis of prostaglandins and thromboxanes, selective inhibitors of inducible cyclooxygenase, selective inhibitors of inducible cyclooxygenase-2, physiologically active substances, paracrine hormones, somatostatin, gastrin, cytokines that mediate interactions involved in humoral and cellular immune responses, formed from lipid physiologically active substances, eicosanoids, β-adrenergic agonists, ipratropium, glucocorticoids, methylxanthines, sodium channel blockers, opioid receptor agonists, calcium channel blockers, membrane stabilizers and leukotriene inhibitors.

Other embodiments of the present invention relate to combinations in which at least one anti-inflammatory compound is an anti-monoclonal antibody (such as eculizumab or pexelizumab), a TNF antagonist such as etanercept, or infliximab, which is an anti-TNF-alpha monoclonal antibody.

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

Other embodiments of the present invention relate to combinations in which at least one active agent is an immunosuppressant, such as an immunosuppressant selected from the group consisting of methotrexate, leflunomide, cyclosporine, tacrolimus, azathioprine, and mycophenolate mofetil.

The disclosed compounds of structure (I) can be administered in combination with other known therapeutic agents, including anticancer agents. As used herein, the term "anticancer agent" refers to any agent administered to a patient with cancer for the treatment of cancer.

In some embodiments, the anticancer agents fall into the following categories:

Alkylating agents: such as altretamine, bendamustine, busulfan, carmustine, chlorambucil, chlormethine, cyclophosphamide, dacarbazine, ifosfamide, improsulfan, tosylate, lomustine, melphalan, mitobronitol, mitolactol, nimustine, ranimustine, temozolomide, thiotepa, treosulfan, mechlorenamine, carboquone; apaziquone, fotemustine, glufosfamide, palifosfamide, pipobroman, trofosfamide, uramustine, TN-3024, VAL-0834;

Platinum compounds: such as carboplatin, cisplatin, eptaplatin, miriplatin hydrate, oxaliplatin, lobaplatin, nedaplatin, picoplatin, satraplatin; lobaplatin, nedaplatin, picoplatin, satraplatin;

DNA-altering agents: such as amrubicin, bisantrene, decitabine, mitoxantrone, procarbazine, trabectedin, clofarabine; amsacrine, brostallicin, pixantrone, laromustine 1,3;

Topoisomerase inhibitors: such as etoposide, irinotecan, razoxan, sobuzoxan, teniposide, topotecan; amonafide, belotecan, elliptinium acetate, voreloxin;

Microtubule modifiers: such as cabazitaxel, docetaxel, eribulin, ixabepilone, paclitaxel, vinblastine, vincristine, vinorelbine, vindesine, vinflunine; fosbretabulin, tesetaxel;

Antimetabolites: such as asparaginase-3, azacitidine, calcium levofolinate, capecitabine, cladribine, cytarabine, enocitabine, floxuridine, fludarabine, fluorouracil, gemcitabine, mercaptopurine, methotrexate, nelarabine, pemetrexed, pralatrexate, azathioprine, thioguanine, carmofur; doxifluridine, elacytarabine, raltitrexed, sapacitabine, tegafur-2,3, trimetrexate;

Anticancer antibiotics: such as bleomycin, dactinomycin, doxorubicin, epirubicin, idarubicin, levamisole, miltefosine, mitomycin C, romidepsin, streptozocin, valrubicin, zinostatin, zorubicin, daunorubicin, plicamycin; aclarubicin, peplomycin, pirarubicin;

Hormones/antagonists: such as abarelix, abiraterone, bicalutamide, buserelin, calusterone, chlorotrianisene, degarelix, dexamethasone, estradiol, fluocortolone, fluoxymesterone, flutamide, fulvestrant, goserelin, histrelin, leuprorelin, megestrol, mitotane, nafarelin, nandrolone, nilutamide, octreotide, prednisolone, raloxifene, tamoxifen, thyrotropin alfa, toremifene, trilostane, triptorelin, diethylstilbestrol; acolbifene, danazol, deslorelin, epitiostanol, orteronel, enzalutamide-1,3;

Aromatase inhibitors: such as aminoglutethimide, anastrozole, exemestane, fadrozole, letrozole, testolactone; formestane;

Small molecule kinase inhibitors: such as 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, mazitinib, midostaurin, motesanib, neratinib, orantinib, perifosine, ponatinib, radotinib, regosertib, tipifamib, tivantinib, tivozanib, trametinib, pimasertib, brivanib alaninate, cediranib.

In some embodiments, drugs administered with the compounds described herein include any suitable drugs advantageously administered by inhalation, such as analgesics, such as codeine, dihydromorphine, ergotamine, fentanyl, or morphine; drugs for the treatment of angina, such as diltiazem; antiallergic drugs, such as cromglycate, ketotifen, or nedocromil; anti-infective drugs, such as cephalosporins, penicillins, streptomycin, sulfonamides, tetracyclines, or pentamidine; antihistamines, such as methapyrilene; anti-inflammatory drugs, such as beclomethasone, flunisolide, budesonide, tipredane, triamcinolone acetonide, or fluticasone; antitussives, such as noscapine; Bronchodilators such as 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 such as amiloride; anticholinergics such as ipratropium, atropine or oxitropium; hormones such as cortisone, hydrocortisone or prednisolone; xanthines such as aminophylline, choline theophylline, lysine theophylline or theophylline; and therapeutic proteins and peptides, such as insulin or glucagon. One skilled in the art will appreciate that, where appropriate, drugs are used in the form of salts (e.g., alkali metal or amine salts or acid addition salts) or esters (e.g., lower alkyl esters) or solvates (e.g., hydrates) to optimize the activity and/or stability of the drug.

The agents disclosed in the present invention, or other suitable agents, are administered depending on the pathological condition being treated. Accordingly, in some embodiments, one or more compounds of the present invention are administered together with other agents described above. When used in combination therapy, the compounds described in the present invention are administered with a second agent simultaneously or separately. This combination administration may include the simultaneous administration of these two agents in a single dosage form, simultaneous administration in different dosage forms, and separate administration. Thus, a compound described in the present invention and any of the agents described above can be formulated together in a single dosage form and administered simultaneously. Alternatively, a compound of the present invention and any of the agents described above can be administered simultaneously, where both agents are contained in different preparations. In another alternative, a compound of the present invention can be administered immediately after any of the agents described above, or vice versa. In some embodiments, in a separate administration protocol, the compound of the present invention and any of the agents described above are administered a few minutes after, or a few hours after, or a few days after.

In some embodiments, the compounds of structure (I) are administered as monotherapy.

To identify signaling pathways or mechanisms and to detect interactions between different signaling pathways, suitable models or model systems have been developed, such as cell culture models and transgenic animal models. Interacting compounds can be used to modulate the signal to determine certain stages of the signaling cascade. Compounds of embodiments of the present invention can also be used as reagents for studying NEK7-dependent signaling pathways in animal and/or cell culture models or in clinical settings for the diseases described herein.

The methods of embodiments of the present invention can be carried out in vitro or in vivo. The susceptibility of a particular cell to treatment with compounds of structure (I) can be determined, in particular, using in vitro assays in a research or clinical setting. Typically, a cell culture is combined with the compound at varying concentrations for a period of time sufficient to inhibit NEK7 activity by the active agent, typically from about 1 hour to one week. In vitro treatment can be carried out using cells grown from a biopsy sample or from a cell line.

In some embodiments, the _IC50 of the compounds of structure (I) for inhibiting NEK7 was determined by the concentration of the compound required to inhibit 50% of the NEK kinase activity. The compounds of structure (I) have activity, as expressed by _IC50 values, of less than about 5 mM, preferably less than about 1 mM, and even more preferably less than about 0.100 mM, as described in more detail in the examples.

The examples and preparations provided below further illustrate and exemplify the compounds of the present invention and methods for preparing and characterizing such compounds. It should be understood that the scope of the present invention is in no way limited by the scope of the following examples and preparations. In the following examples and in the present description and claims, molecules with one stereocenter, unless otherwise indicated, exist as a racemic mixture. Molecules with two or more stereocenters, unless otherwise indicated, exist as a racemic mixture of diastereoisomers. Individual enantiomers/diastereoisomers can be prepared by methods known to those skilled in the art.

Examples

The following examples are given as typical.

General techniques

All proton NMR experiments were performed on a Bruker NEO spectrometer equipped with a BBFO probe at 400 MHz. Deuterated solvents contained less than 0.05% v/v tetramethylsilane, which was used as a reference signal (set at 0.00 ppm). If the deuterated solvents did not contain tetramethylsilane, residual non-deuterated solvent peaks were used as a reference signal according to 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). Splitting characteristics describe apparent multiplicities and are designated as s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), qt (quintuplet), or brs (broad singlet).

LC/MS analyses were performed using an Agilent Technologies UHPLC 1290 Infinity II with a G6125 MS detector.

Microwave heating reactions were carried out in a Monowave 300 apparatus from Anton Paar GmbH according to standard protocols.

Study of the NEK7 enzyme

Casein substrate (from cow's milk, a hydrolyzed and partially dephosphorylated mixture of caseins α, β, and κ, obtained from Sigma Aldrich, catalog # C4765, was 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 using an N-terminal GST tag, obtained from SignalChem, catalog # N09-10G, 0.1 μg/μL) were mixed in assay buffer (20 mM Hepes pH 7.5, 10 mM MgCl ₂ , 1 mM EGTA, 0.02% Brij35, 0.02 mg/mL BSA, 0.1 mM Na ₃ VO ₄ , 2 mM DTT, 1% DMSO). Test compounds (serial 3-fold dilutions in DMSO from 10 μM to 0.5 nM) or diluent (1% DMSO) were dosed into the kinase reaction mixture using Acoustic technology (Echo550; nanoliter range). After incubation at room temperature for 20 min, the kinase reaction was initiated by the addition of [ ^33P ]-ATP (specific activity 10 μCi/μL) and the mixture was incubated at room temperature for 2 h. The reaction was then stopped by dropping the reaction mixture onto strips of P81 phosphocellulose paper. After washing, the radioactivity of the P81 paper was measured and kinase activity data were presented as the percentage of remaining kinase activity in the test samples compared to reactions in diluent. IC ₅₀ values and curve fitting were performed using Prism (GraphPad Software).

IL-Iβ release assay

Approximately 1.5 million THP-1 cells were plated into each well of a 6-well TC plate and incubated with 40 nM PMA in RPMI (10% FBS, 1% Penstrep) for 24 h. The media were then removed and the cells were left in RPMI (10% FBS, 1% Penstrep) for 24 h, and then the media were removed and the cells were pretreated for 2 h with the test compounds at different concentrations (typically serial 3-fold dilutions in RPMI+5% FBS, concentrations ranging from 1 μM to 0.5 nM) in RPMI (5% FBS). The media were again removed, and the cells were incubated with 250 ng/ml LPS and test compounds (concentrations as above) in RMPI (5% FBS) for 2 h. The media was removed a final time, and the cells were incubated with 20 μM nigericin and test compounds (concentrations as above) in Opti-MEM for 30 min. The cell media were then collected, and the amount of cleaved IL-1β was determined using a JESS instrument (Protein Simple) using standard protocols. Cleaved IL-1β antibodies were purchased from Cell Signaling (catalogue #83186S) and used at a 1:20 dilution in antibody diluent 2. 1× anti-rabbit HRP secondary antibody from Protein Simple, along with Protein Simple luminol and peroxide, was used for chemiluminescent detection. The incubation time of primary antibodies was increased from 30 min to 60 min.

Abbreviations:

°C (degrees Celsius); ¹ H NMR (proton nuclear magnetic resonance); ACN (acetonitrile); Boc (tert-butyloxycarbonyl); DCM (dichloromethane); DIPEA (N,N-diisopropylethylamine); DMAP (4-dimethylaminopyridine); DMF (N,N-dimethylformamide); DMSO-d ₆ (deuterated dimethyl sulfoxide); eq. (equivalent); EtOAc (ethyl acetate); g (gram); h (hour); HPLC (high performance liquid chromatography); LCMS (liquid chromatography mass spectrometry); MeOH (methanol); mg (milligram); min (min); ml (milliliter); mmol (millimole); n-BuOH (1-butanol); Pd(PPh ₃ ) ₄ (tetrakis(triphenylphosphine)palladium); PdCl ₂ (dppf) ([1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride); TBAF (tetra-n-butyl ammonium fluoride); TBDMS (tert-butyldimethylsilyl); TFA (trifluoroacetic acid); THF (tetrahydrofuran); TLC (thin layer chromatography)

Obtaining synthetic intermediates

Intermediate product A

4-chloro-5-iodo-7H-pyrrole o[2,3-d]pyrimidine

N-iodosuccinimide (1.465 g, 6.51 mmol) was added with stirring to a solution of 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1.000 g, 6.51 mmol) in DMF (10 mL) at 0 °C and the resulting mixture was stirred at 25 °C for 12 h. After the reaction was complete (as determined by TLC), the reaction mixture was poured into ice-cold water (100 mL) and stirred at 25 °C for 15 min. The resulting solid was filtered, washed with water (2 x 25 mL), and dried to give the title compound as an off-white solid (1.7 g, 93% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=12.96 (bs, 1H), 8.60 (s, 1H), 7.95 (d, J=2.40 Hz, 1H); LCMS: 279.9 [M+H].

Intermediate product B1

4-chloro-7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidine

Copper(II) acetate (0.650 g, 3.58 mmol), 2,2'-bipyridine (0.559 g, 3.58 mmol) and sodium bicarbonate (0.601 g, 7.16 mmol) were added to a solution of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (A1, 1.000 g, 3.58 mmol) and cyclopropylboronic acid (0.615 g, 7.16 mmol) in dichloroethane (10 mL) and the resulting mixture was stirred at 70 °C under oxygen atmosphere for 12 h. After completion of the reaction (as determined by TLC), the reaction mixture was filtered through a pad of celite, which was then washed with DCM (2×20 mL). The combined filtrates were washed with water (20 mL) and brine (25 mL), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give a crude material that was purified by flash chromatography (silica gel 230-400 mesh eluting with 15% EtOAc in petroleum ether) to afford the title compound as an off-white solid (0.7 g, 61% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.67 (s, 1H), 7.96 (s, 1H), 3.63-3.69 (m, 1H), 1.06-1.10 (m, 4H). LCMS: 319.9 [M+H]. Intermediate B2

4-chloro-5-iodo-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidine

_K2CO3 (0.40 g, 2.86 mmol) and 3-iodoxetane (0.32 g _, 1.71 mmol) were added to a solution of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (A, 0.40 g, 1.43 mmol) in DMF (5 mL) and the resulting mixture was stirred at 90 °C for 16 h in a sealed tube. After the reaction was complete (as determined by TLC), the reaction mixture was poured into 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 (0.2 g, 42% yield). LCMS: 335.7 [M+H].

Intermediate product B3

1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol

_NaH2PO4 (0.105 g, 0.877 mmol) was added to a mixture _of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (A1, 0.250 g, 0.895 mmol), 2,2-dimethyloxirane (0.157 mL, _1.762 mmol), and _K2CO3 (0.121 g, 0.877 mmol) in ACN (3 mL) and water (1 mL). The resulting mixture was microwaved at 150°C for 1 h in a sealed tube. After the reaction was complete (as determined by TLC), the reaction mixture was concentrated under reduced pressure and the resulting crude material was purified by flash chromatography (silica gel 230-400 mesh eluting with 18% EtOAc in petroleum ether) to afford the title compound as a pale brown solid (0.1 g, 17% yield). LCMS: 351.9 [M+H].

Intermediate product B4

4-chloro-5-iodo-7-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidine

Triethylamine (0.905 g, 8.95 mmol) and copper(II) acetate (0.975 g, 5.37 mmol) were added to a solution of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (A, 1.000 g, 3.58 mmol) and 3-pyridylboronic acid (0.880 g, 7.16 mmol) in DCM (25 mL) and the resulting mixture was stirred at 40 °C under oxygen for 40 h. After the reaction was complete (as assessed by LCMS), the reaction mixture was filtered through a pad of celite, which was then washed with DCM (2 x 50 mL). The combined filtrates were washed with water (10 mL) and brine (10 mL), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give the crude material. It was stirred with 30% diethyl ether in petroleum ether for 30 min at 25°C, filtered and dried to give the title compound as a brown solid (0.4 g, 29% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 9.05 (bs, 1H), 8.73 (s, 1H), 8.67 (bs, 1H), 8.48 (s, 1H), 8.26-8.28 (m, 1H), 7.64-7.67 (m, 1H). LCMS: 356.8 [M+H].

Intermediate product B5

4-chloro-5-iodo-7-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidine

The title compound was prepared by a procedure similar to that described for B4 using 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (A, 0.50 g, 1.789 mmol) and 4-pyridylboronic acid (0.44 g, 3.580 mmol) as starting materials and was obtained as a brown solid (0.21 g, 29% yield). LCMS: 356.9 [M+H].

Intermediate product B6

4-chloro-5-iodo-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidine

The title compound was prepared as described in PCT Publication No. WO 2017/220477.

Intermediate product B7

7-(3-(benzyloxy)cyclobutyl)-4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine

_Cs2CO3 (0.583 g, 1.789 mmol) and 3-(benzyloxy)cyclobutyl methanesulfonate (prepared as described in PCT Publication No. WO 2019/092170, 0.459 g _, 1.789 mmol) were added to a solution of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (A, 0.250 g, 0.895 mmol) in DMF (5 mL) and the resulting mixture was stirred at 90 °C for 12 h. After the reaction was complete (as determined by TLC), the reaction mixture was poured onto ice-water (50 mL) and extracted with ethyl acetate (2×30 mL). The combined organic extracts _{were dried over Na2SO4 ,} filtered, and concentrated under reduced pressure to give a crude material, which was purified by flash chromatography (silica gel 230-400 mesh eluting with 30% EtOAc in petroleum ether) to afford the title compound as a colorless gum (0.14 g, 31% yield). LCMS: 440.0 [M+H].

Intermediate product B8

2-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-ol

_K2CO3 (0.742 g, 5.37 mmol) and 2-bromoethan-1-ol (0.537 g _, 4.29 mmol) were added to a solution of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (A, 1.000 g, 3.58 mmol) in DMF (6 mL) and the resulting suspension was stirred at 80 °C for 2 h. After the reaction was complete (as determined by TLC), the reaction mixture was poured onto crushed ice (25 g). The resulting solid was filtered, washed with water (20 mL) and dried to give the title compound as a yellow solid (0.84 g, 64% yield). LCMS: 323.9 [M+H].

Intermediate product B9

4-chloro-1-cyclopropyl-3-iodo-1H-pyrrolo[3,2-c]pyridine Step 1: Synthesis of 4-chloro-1-cyclopropyl-1H-pyrrolo[3,2-c]pyridine

Triethylamine (0.332 g, 3.280 mmol), copper(II) acetate (0.298 g, 1.638 mmol), and molecular sieves (powdered, 0.050 g) were added to a solution of 4-chloro-1H-pyrrolo[3,2-c]pyridine (0.250 g, 1.638 mmol) and cyclopropylboronic acid (0.279 g, 3.280 mmol) in DMF (10 mL), and the resulting suspension was stirred at 60 °C for 12 h in a sealed tube. After the reaction was complete (as determined by TLC), the reaction mixture was filtered through a pad of celite, which was then washed with EtOAc. The combined filtrates were concentrated under reduced pressure to give a crude material, which was purified using Isolera (silica gel 230-400 mesh eluting with 20% EtOAc in petroleum ether) to afford the title compound as a yellow solid (0.19 g, 59% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.04 (d, J = 5.6 Hz, 1H), 7.56-7.60 (m, 2H), 6.52-6.53 (m, 1H), 3.55-3.58 (m, 1H), 1.00-1.13 (m, 4H). LCMS: 193.1 [M+H].

Step 2: Synthesis of 4-chloro-1-cyclopropyl-3-iodo-1H-pyrrolo[3,2-c]pyridine

N-iodosuccinimide (0.350 g, 1.557 mmol) was added to a solution of 4-chloro-1-cyclopropyl-1H-pyrrolo[3,2-c]pyridine (0.200 g, 1.038 mmol) in DMF (5 mL) and the resulting mixture was stirred at 80 °C for 1 h. After the reaction was complete (as determined by LCMS), the reaction mixture was poured into crushed ice (25 g) and extracted with EtOAc (2×25 mL). The combined organic extracts were washed with brine, dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give the title product (0.2 g), which was used without further purification. LCMS: 319.0 [M+H].

Intermediate product C1

7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine

A mixture of 4-chloro-7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (B1, 1.00 g, 2.191 mmol) and ammonium hydroxide (25% in water, 5 mL) was heated under microwave irradiation at 150 °C for 1 h. After the reaction was complete (as determined by TLC), the reaction mixture was concentrated under reduced pressure to give the title compound as an off-white solid (0.75 g, 80% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.12 (s, 1H), 7.39 (s, 1H), 6.57 (bs, 2H), 3.48-3.54 (m, 1H), 0.97-1.01 (m, 4H). LCMS: 301.0 [M+H].

Intermediate product C2

5-iodo-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that used for Cl using 4-chloro-5-iodo-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidine (B2, 0.5 g, 1.49 mmol) and aqueous ammonium hydroxide (25% in water, 2.5 mL) as starting materials and was obtained as a pale brown solid (0.27 g, 58% yield). LCMS: 316.8 [M+H].

Intermediate product C3

1-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol

The title compound was prepared by a procedure similar to that described for Cl using 1-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol (B3, 0.1 g, 0.284 mmol) and ammonium hydroxide (25% in water, 0.5 mL) as an off-white solid (0.08 g, 85% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.12 (s, 1H), 7.38 (s, 1H), 4.81 (s, 1H), 4.04 (s, 2H), 1.03 (s, 6H). LCMS: 333.0 [M+H].

Intermediate product C4

5-iodo-7-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

Ammonium hydroxide (25% in water, 1 mL) was added to a solution of 4-chloro-5-iodo-7-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidine (B4, 0.30 g, 0.841 mmol) in dioxane (10 mL), and the resulting mixture was heated in a microwave at 150 °C for 2 h. After the reaction was complete (as determined by LCMS), the reaction mixture was concentrated under reduced pressure to give a crude material, which was washed with methyl tert-butyl ether and dried to give the title compound as an off-white solid (0.21 g, 63% yield). LCMS: 337.8 [M+H]. Intermediate C5

5-iodo-7-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for C4 using 4-chloro-5-iodo-7-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidine (B5, 0.21 g, 0.589 mmol) and ammonium hydroxide (25% in water, 1 mL) as starting materials and was obtained as an off-white solid (0.16 g, 69% yield). LCMS: 337.9 [M+H].

Intermediate product C6

5-iodo-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared as described in PCT Publication No. WO 2017/220477.

Intermediate product C7

7-(3-(benzyloxy)cyclobutyl)-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for C4 using 7-(3-(benzyloxy)cyclobutyl)-4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (B7, 0.140 g, 0.318 mmol) and ammonium hydroxide (25% in water, 1.4 mL) as starting materials and was obtained as an off-white solid (0.06 g, 45% yield). LCMS: 421.1 [M+H].

Intermediate product C8

5-iodo-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared as described in PCT Publication No. WO 2014/184069 A1.

Intermediate product C9

2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-ol

The title compound was prepared by a procedure similar to that described for C4 using 2-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-ol (B8, 0.84 g, 2.61 mmol) and ammonium hydroxide (25% in water, 8 mL) as starting materials and was obtained as an off-white solid (0.94 g, 69% yield). LCMS: 305.0 [M+H].

Intermediate product C10

7-cyclobutyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared as described in PCT Publication No. WO 2016/075224

Intermediate product C11

5-iodo-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared as described in PCT Publication No. WO 2016/075224.

Intermediate product D1

5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

A mixture of 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (Cl, 0.160 g, 0.533 mmol), 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.190 g, 0.800 mmol) and _K2CO3 (0.221 g, 1.599 mmol) in 1,4-dioxane (1 mL) and water (0.3 mL) was purged with _N2 for 10 _min . Then Pd( _PPh3 ) ₄ (0.062 g, 0.053 mmol) was added and the reaction mixture was stirred at 100°C for 12 h. After the reaction was complete (as determined by TLC), the mixture was filtered through a pad of Celite, which was then washed with EtOAc (2×10 mL). The combined filtrates were concentrated under reduced pressure to give a crude material, which was purified by flash chromatography (silica gel 230-400 mesh eluting with 3% MeOH in DCM) to give the title compound as a yellow solid (0.110 g, 73% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=8.14 (s, 1H), 7.13 (s, 1H), 7.05-7.09 (m, 1H), 6.95-6.98 (m, 1H), 6.82-6.86 (m, 1H), 6.10 (bs, 2H), 5.22 (bs, 2H), 3.52-3.58 (m, 1H), 1.00-1.04 (m, 4H). LCMS: 284.1 [M+H].

Intermediate product D2

5-(4-aminophenyl)-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

Step 1: Synthesis of 5-(4-nitrophenyl)-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that used for D1 using 4-chloro-5-iodo-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidine (C2, 0.252 g, 0.797 mmol) and (4-nitrophenyl)boronic acid (0.200 g, 1.198 mmol) as starting materials and was obtained as a pale brown solid (0.143 g, 58% yield). LCMS: 312.1 [M+H].

Step 2: Synthesis of 5-(4-aminophenyl)-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

Iron powder (0.251 g, 4.5 mmol) and ammonium chloride (0.240 g, 4.5 mmol) were added to a solution of 5-(4-nitrophenyl)-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (0.14 g, 0.45 mmol) in ethanol (5 mL) and water (2 mL), and the resulting mixture was stirred at 80 °C for 3 h. After the reaction was complete (as determined by TLC), the reaction mixture was filtered through a pad of celite, which was then washed with EtOAc (2×5 mL). The combined filtrates were concentrated under reduced pressure to give a residue which was dissolved in EtOAc (25 mL), washed with brine ( ₅ mL), dried over _Na2SO4 , filtered and concentrated under reduced pressure to give the title compound as a brown solid (0.12 g, quantitative yield), which was used without further purification. LCMS: 281.9 [M+H].

Intermediate product D3

1-(4-amino-5-(4-amino-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol

The title compound was prepared by a procedure similar to that used for D1 using 1-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol (C3 (0.100 g, 0.301 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.086 g, 0.361 mmol) as starting materials and was obtained as a pale yellow resinous substance (0.05 g, 53% yield). LCMS: 316.1 [M+H].

Intermediate product D4

5-(4-aminophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

Step 1: Synthesis of 7-cyclopropyl-5-(4-nitrophenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared using a procedure similar to that described for D1 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C1, 0.18 g, 0.60 mmol) and (4-nitrophenyl)boronic acid (0.12 g, 0.72 mmol) as starting materials and was obtained as a pale brown solid (0.10 g, 56% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=8.28-8.32 (m, 2H), 8.21 (s, 1H), 7.77 (s, 1H), 7.70-7.73 (m, 2H), 7.55 (s, 1H), 5.69 (bs, 2H), 3.61-3.64 (m, 1H), 1.04-1.09 (m, 4H). LCMS: 296.1 [M+H].

Step 2: Synthesis of 5-(4-aminophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described in step 2 of D2 using 7-cyclopropyl-5-(4-nitrophenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (0.10 g, 0.33 mmol) and Fe/NH ₄ Cl as starting materials and was obtained as a brown resinous substance (0.08 g, 90% yield), which was used without further purification. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=8.13 (s, 1H), 7.04-7.11 (m, 2H), 7.04 (s, 1H), 6.63-6.67 (m, 2H), 6.05 (bs, 2H), 5.27 (bs, 2H), 3.51-3.57 (m, 1H), 0.99-1.04 (m, 4H). LCMS: 266.0 [M+H].

Intermediate product D5

5-(4-amino-2-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D1 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C1, 0.21 g, 0.69 mmol) and 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.20 g, 0.83 mmol) as starting materials and was obtained as a pale yellow resinous substance (0.15 g, 76% yield). LCMS: 284.1 [M+H].

Intermediate product D6

5-(6-aminopyridin-3-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

Step 1: Synthesis of 7-cyclopropyl-5-(6-nitropyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D1 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C1, 0.28 g, 0.94 mmol) and 2-nitro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (0.28 g, 1.13 mmol) as starting materials and was obtained as a pale brown solid (0.16 g, 57% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=8.73 (d, J=2.0 Hz, 1H), 8.39 (d, J=8.8 Hz, 1H), 8.23 (s, 1H), 8.17-8.20 (m, 1H), 7.67 (s, 1H), 6.49 (bs, 2H), 3.61-3.67 (m, 1H), 1.06-1.09 (m, 4H). LCMS: 297.1 [M+H].

Step 2: Synthesis of 5-(6-aminopyridin-3-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for step 2 in D2 using 7-cyclopropyl-5-(6-nitropyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (0.16 g, 0.54 mmol) and Fe/NH ₄ Cl as a pale brown solid (0.1 g, 70% yield) which was used without further purification. LCMS: 267.0 [M+H].

Intermediate product D7

5-(4-aminocyclohex-1-en-1-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

Step 1: Synthesis of tert-butyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)cyclohex-3-en-1-yl)carbamate

_K2CO3 (0.318 g, 2.299 mmol) was added to a solution of 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (Cl _, 0.230 g, 0.766 mmol) and tert-butyl (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)carbamate (0.372 g, 1.150 mmol) in dioxane (1 mL) and water (0.3 mL). The solution was purged with _N2 for 10 min, then Pd( _PPh3 ) ₄ (0.044 g, 0.038 mmol) was added and the resulting mixture was heated in the microwave at 100°C for 1 h. After the reaction was complete (as determined by TLC), the reaction mixture was filtered through a pad of celite, which was then washed with EtOAc (2×10 mL). The combined filtrates were concentrated under reduced pressure to give the crude material, which was purified by preparative HPLC (mass detection, eluting with a gradient of ammonium acetate in water and ACN) to give the desired product as a pale yellow gum (0.18 g, 62% yield). LCMS: 370.2 [M+H].

Step 2: Synthesis of 5-(4-aminocyclohex-1-en-1-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

TFA (0.012 g, 0.108 mmol) was added to a solution of tert-butyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)cyclohex-3-en-1-yl)carbamate (0.040 g, 0.108 mmol) in DCM (2 mL) at 0 °C and the resulting solution was stirred at room temperature for 12 h. After completion of the reaction (as determined by TLC), the reaction mixture was concentrated under reduced pressure to give the title product as a brown gum (0.029 g), which was used without further purification. LCMS: 270.1 [M+H].

Intermediate product D8

5-(4-amino-3-fluorophenyl)-7-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

A mixture of 5-iodo-7-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C4, 0.160 g, 0.475 mmol), 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2- _dioxaborolan -2-yl)aniline (0.169 g, 0.712 mmol) and _K2CO3 (0.131 g, 0.949 mmol) in dioxane (5 mL), water (2 mL) and ethanol (3 mL) was purged with _N2 for 10 min. _PdCl2 (dppf) (0.017 g, 0.024 mmol) was added and the resulting mixture was heated in the microwave at 100 °C for 1 h. After the reaction was complete (as assessed by LCMS), the reaction mixture was filtered through a pad of celite, which was then washed with EtOAc (5 mL). The combined filtrates were concentrated under reduced pressure to give a residue, which was taken up in EtOAc (50 mL), washed with water (5 mL) and brine (5 mL ₎ , dried over _Na2SO4 , filtered, and concentrated under reduced pressure. The resulting crude material was purified by GRACE (silica gel 230-400 mesh eluting with 4% MeOH in DCM) to afford the title compound as a brown solid (0.2 g, 70% yield). LCMS: 321.0 [M+H].

Intermediate product D9

5-(4-amino-3-fluorophenyl)-7-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 5-iodo-7-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C5, 0.160 g, 0.475 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.169 g, 0.712 mmol) as starting materials and was obtained as a brown solid (0.08 g, 51% yield). LCMS: 321.0 [M+H].

Intermediate product D10

5-(4-amino-3-fluorophenyl)-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 5-iodo-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C6, 0.180 g, 0.504 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.131 g, 0.554 mmol) as a brown resinous substance (0.15 g, 80% yield). LCMS: 341.1 [M+H].

Intermediate product D11

5-(4-amino-3-methylphenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C1, 0.250 g, 0.833 mmol) and 2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.214 g, 0.916 mmol) as starting materials and was obtained as a brown resinous substance (0.13 g, 56% yield). LCMS: 280.1 [M+H].

Intermediate product D12

5-(4-amino-3-fluorophenyl)-7-(3-(benzyloxy)cyclobutyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 7-(3-(benzyloxy)cyclobutyl)-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C7, 0.060 g, 0.143 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.037 g, 0.157 mmol) as starting materials and was obtained as a brown solid (0.03 g, 53% yield). LCMS: 404.2 [M+H].

Intermediate product D13

Step 1: Synthesis of 7-cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

Potassium acetate (0.245 g, 2.499 mmol) was added to a solution of 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (Cl, 0.250 g, 0.833 mmol) and bis(pinacolato)diboron (0.317 g, 1.250 mmol) in DMSO (5 mL) and the resulting mixture was purged with N ₂ for 10 min. PdCl ₂ (dppf) (0.030 g, 0.042 mmol) was then added and the reaction mixture was stirred at 85 °C for 2 h. After completion of the reaction, the reaction mixture was filtered through a pad of celite, which was then washed with DCM (2 x 20 mL). The combined filtrates were concentrated under reduced pressure to give the title compound as a black residue, which was used without further purification. LCMS: 300.9 [M+H].

Step 2: Synthesis of 5-(5-amino-4-methylpyridin-2-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 6-bromo-4-methylpyridin-3-amine (0.142 g, 0.759 mmol) and 7-cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (0.251 g, 0.835 mmol) as starting materials and was obtained as a brown gum (0.05 g, 13% yield). LCMS: 281.0 [M+H].

Intermediate product D14

5-(5-aminopyridin-2-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 6-bromo-4-methylpyridin-3-amine (0.130 g, 0.751 mmol) and 7-cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (step 1 for intermediate D13, 0.248 g, 0.827 mmol) as starting materials and was obtained as a brown gum (0.03 g, 4.5% yield). LCMS: 267.0 [M+H].

Intermediate product D15

5-(4-amino-3,5-difluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C1, 0.500 g, 0.751 mmol) and 2,6-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.425 g, 1.666 mmol) as starting materials and was obtained as a pale yellow solid (0.10 g, 19% yield). LCMS: 302.1 [M+H].

Intermediate product D16

5-(4-amino-2,5-difluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (Cl, 0.125 g, 0.417 mmol) and 2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (prepared as described in PCT Publication No. WO 2017/172093, 0.106 g, 4.17 mmol) as starting materials and was obtained as a pale yellow solid (0.05 g, 40% yield). LCMS: 302.1 [M+H].

Intermediate product D17

5-(4-amino-2,6-difluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (Cl, 0.055 g, 0.183 mmol) and 3,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (prepared as described in PCT Publication No. WO 2017/172093, 0.056 g, 0.220 mmol) as a pale brown gum (0.046 g), which was used without further purification. LCMS: 301.9 [M+H].

Intermediate product D18

1-(4-amino-5-(6-aminopyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol

The title compound was prepared by a procedure similar to that described for D8 using 1-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol (C3, 0.280 g, 0.733 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (0.161 g, 0.733 mmol) as starting materials and was obtained as a yellow resinous substance (0.12 g, 50% yield). LCMS: 299.1 [M+H].

Intermediate product D19

5-(2-aminopyrimidin-5-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C1, 0.25 g, 0.833 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (0.184 g, 0.833 mmol) as starting materials and was obtained as a colorless resinous substance (0.08 g, 34% yield). LCMS: 268.2 [M+H].

Intermediate product D20

(2-amino-5-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)methanol

The title compound was prepared by a procedure similar to that described for D8 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (Cl, 0.200 g, 0.666 mmol) and (2-amino-5-(4,4,5,5-tetramethyl-1,3-dioxolan-2-yl)phenyl)methanol (prepared as described in PCT Publication No. WO 2011/130628, 0.184 g, 0.733 mmol) as a pale yellow gum (0.025 g, 13% yield). LCMS: 296.0 [M+H].

Intermediate product D21

2-amino-5-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)benzonitrile

The title compound was prepared by a procedure similar to that described for D8 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C1, 0.250 g, 0.833 mmol) and 2-amino-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (0.203 g, 0.833 mmol) as starting materials and was obtained as a yellow resinous substance (0.13 g, 34% yield). LCMS: 291.2 [M+H].

Intermediate product D22

5-(4-amino-3-fluorophenyl)-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 5-iodo-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C8, 0.200 g, 0.629 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.149 g, 0.629 mmol) as starting materials and was obtained as a pale brown solid (0.12 g, 42% yield). LCMS: 302.2 [M+H].

Intermediate product D23

2-(4-amino-5-(4-amino-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-ol

The title compound was prepared by a procedure similar to that described for D8 using 2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-ol (C9, 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) as starting materials and was obtained as a brown solid (0.3 g, 63% yield). LCMS: 288.1 [M+H].

Intermediate product D24

5-(4-amino-3-fluorophenyl)-7-cyclobutyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 7-cyclobutyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C10, 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) as starting materials and was obtained as a brown solid (0.18 g, 37% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) 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), 2.67-2.68 (m, 2H), 2.38-2.39 (m, 2H), 1.85-1.86 (m, 2H). LCMS: 298.0 [M+H].

Intermediate product D25

5-(4-amino-3-chlorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (Cl, 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) as starting materials and was obtained as a yellow solid (0.03 g, 12% yield). LCMS: 300.1 [M+H].

Intermediate product D26

5-(4-amino-3-methoxyphenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 7-cyclopropyl-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C1, 0.250 g, 0.833 mmol) and 2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.208 g, 0.833 mmol) as starting materials and was obtained as a pale yellow resinous substance (0.04 g, 16% yield). LCMS: 296.1 [M+H].

Intermediate product D27

5-(4-amino-3-fluorophenyl)-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

The title compound was prepared by a procedure similar to that described for D8 using 5-iodo-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (C11, 0.155 g, 0.452 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.107 g, 0.452 mmol) as starting materials and was obtained as a brown solid (0.084 g, 27% yield). LCMS: 327.2 [M+H].

Intermediate product E1-E25

General procedure for the synthesis of carbamate intermediates E:

Pyridine (1.2 equiv) and phenyl chloroformate (1.5 equiv) were added to a solution of the amine (1.0 equiv) in THF (10 vol) at 0 °C. The reaction mixture was allowed to warm to 25 °C and stirred for 12 h. After the reaction was complete (as determined by TLC), the mixture was diluted with EtOAc (10 mL) and washed with brine (5 mL). The organic layer was dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give a crude material, which was purified by flash chromatography (silica gel 230-400 mesh eluting with 10 to 20% EtOAc in petroleum ether) to afford the desired carbamate.

The following carbamates were prepared according to the general procedure described above:

All amines used for the synthesis of carbamate intermediates E are commercially available except the following:

3-(1-(Trifluoromethyl)cyclopropyl)isoxazol-5-amine (precursor for E6) and 5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-amine (precursor for E7) were synthesized as described in Synthesis 2013, 45, 171-173.

3-(1,1,1-Trifluoro-2-methylpropan-2-yl)isoxazol-5-amine (precursor for E8) and 3-(2-fluoropropan-2-yl)isoxazol-5-amine (precursor for E9) were synthesized from methyl 3,3,3-trifluoro-2,2-dimethylpropanoate and methyl 2-fluoro-2-methylpropionate, respectively, then according to the procedure described in Synthesis 2013, 45, 171-173.

3-(1-((Tert-butyldimethylsilyl)oxy)-2-methylpropan-2-yl)isoxazol-5-amine (precursor for E16) was synthesized as described in PCT Publication No. WO 2010/036630.

2-(5-Aminoisoxazol-3-yl)-2-methylpropanenitrile (precursor for E22) and 3-(1-(trifluoromethyl)cyclobutyl)isoxazol-5-amine (precursor for E24) were synthesized as described in J. Med. Chem. 2012, 55(3), 1082-1105

3-(((Tert-butyldiphenylsilyl)oxy)methyl)isoxazol-5-amine (precursor for E23) was synthesized as described in PCT Publication No. WO 2013/104561.

5-(1-(Trifluoromethyl)cyclobutyl)isoxazol-3-amine (precursor for E25) was synthesized as described in PCT Publication No. WO 2011/022473.

Synthesis of 3-(3-methyloxetan-3-yl)isoxazol-5-amine (precursor for E14):

NH ₂ OH⋅H ₂ SO ₄ (0.520 g, 3.16 mmol) was added to a solution of 3-(3-methyloxetan-3-yl)-3-oxopropanenitrile (prepared as described in PCT Publication No. WO 2019/192962, 0.400 g, 2.87 mmol) and sodium hydroxide (0.126 g, 3.16 mmol) in EtOH (10 mL) and water (10 mL). The pH of the resulting mixture was adjusted to 7.5 with aqueous NaOH (1 M) and the reaction mixture was stirred at 80°C for 15 h. After the reaction was complete (as determined by TLC), the reaction mixture was concentrated under reduced pressure to give a residue, which was taken up in EtOAc (25 mL), washed with water (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The resulting crude material was purified by flash chromatography (silica gel 230-400 mesh eluting with 30% EtOAc in petroleum ether) to afford the title product as a pale brown solid (0.09 g, 20% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ = 5.21 (s, 1H), 4.90-4.93 (m, 2H), 4.56-4.59 (m, 2H), 1.70 (s, 3H). LCMS: 155.1 [M+H].

Synthesis of 3-(1-methylcyclobutyl)isoxazol-5-amine (precursor for E21):

NH ₂ OH⋅H ₂ SO ₄ (0.699 g, 4.25 mmol) was added to a solution of 3-(1-methylcyclobutyl)-3-oxopropanenitrile (prepared as described in PCT Publication No. WO 2017/060874, 0.500 g, 3.86 mmol) and sodium hydroxide (0.170 g, 4.25 mmol) in EtOH (10 mL) and water (10 mL). The pH of the resulting mixture was adjusted to 7.5 with aqueous NaOH (1 M) and the reaction mixture was stirred at 80°C for 15 h. After the reaction was complete (as determined by TLC), the reaction mixture was concentrated under reduced pressure to give a residue, which was taken up in DCM (25 mL), washed with water (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The resulting crude material was purified by flash chromatography (silica gel 230-400 mesh eluting with 40% EtOAc in petroleum ether) to afford the title product as an off-white solid (0.110 g, 19% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ = 5.04 (s, 1H), 2.43-2.49 (m, 2H), 1.96-2.02 (m, 4H), 1.50 (s, 3H). LCMS: 153.2 [M+H].

Obtaining example connections

General procedure for obtaining urea for the synthesis of compounds of examples 1-62

Method A - Triethylamine (2.0 equiv) was added to a mixture of intermediate amine D (1.0 equiv) and intermediate carbamate E (1.0 equiv) in THF (10 vol), and the resulting mixture was stirred at 60°C for 12 h in a sealed tube. After completion of the reaction (as determined by LCMS), the reaction mixture was concentrated under reduced pressure to yield a crude material, which was purified by preparative reverse-phase HPLC to afford the desired product.

Method B - DMAP (0.05 equiv) and DIPEA (1.5 equiv) were added to a solution of intermediate amine D (1.0 equiv) and intermediate carbamate E (1.0 equiv) in THF (10 vol), and the resulting mixture was stirred at 60°C for 12 h in a sealed tube. After completion of the reaction (as determined by LCMS), the reaction mixture was concentrated under reduced pressure to yield a crude material, which was purified by preparative reverse-phase HPLC to afford the desired product.

The following compounds were prepared using the general procedures outlined above.

Example 1

1-(4-(4-amino-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-3-(3-(tert-butyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-aminophenyl)-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D2, 0.178 g, 0.63 mmol) and phenyl (3-(tert-butyl)isoxazol-5-yl) carbamate (E1, 0.16 g, 0.30 mmol) as starting materials and was obtained as an off-white solid (0.026 g, 9% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.48 (bs, 1H), 9.34 (bs, 1H), 8.15 (s, 1H), 7.70 (s, 1H), 7.61 (d, J=8.8 Hz, 2H), 7.45 (d, J=8.8 Hz, 2H), 6.08 (bs, 2H), 6.07 (s, 1H), 5.86-5.90 (m, 1H), 4.97-5.05 (m, 4H), 1.27 (s, 9H). LCMS: 448.2 [M+H].

Example 2

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(tert-butyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-aminophenyl)-7-(oxetan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D2, 0.178 g, 0.63 mmol) and phenyl-(3-(tert-butyl)isoxazol-5-yl)carbamate (E1, 0.16 g, 0.30 mmol) as starting materials and was obtained as an off-white solid (0.026 g, 9% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.48 (bs, 1H), 9.34 (bs, 1H), 8.15 (s, 1H), 7.70 (s, 1H), 7.61 (d, J=8.8 Hz, 2H), 7.45 (d, J=8.8 Hz, 2H), 6.08 (bs, 2H), 6.07 (s, 1H), 5.86-5.90 (m, 1H), 4.97-5.05 (m, 4H), 1.27 (s, 9H). LCMS: 448.2 [M+H].

Example 3

1-(5-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pyridin-2-yl)-3-(3-(tert-butyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(6-aminopyridin-3-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D6, 0.100 g, 0.376 mmol) and phenyl (3-(tert-butyl)isoxazol-5-yl) carbamate (E1, 0.098 g, 0.376 mmol) as starting materials and was obtained as an off-white solid (9.6 mg, 6% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.46-8.47 (m, 1H), 8.22 (s, 1H), 7.89-7.92 (m, 2H), 7.35-7.38 (m, 1H), 7.28 (s, 1H), 6.23 (s, 1H), 3.50-3.57 (m, 1H), 1.27 (s, 9H), 1.07-1.16 (m, 4H); LCMS: 433.2 [M+H].

Example 4

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.100 g, 0.35 mmol) and phenyl (5-(tert-butyl)isoxazol-3-yl)carbamate (E2, 0.091 g, 0.35 mmol) as starting materials and was obtained as an off-white solid (0.021 g, 13% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=9.86 (bs, 1H), 8.87 (bs, 1H), 8.17-8.21 (m, 2H), 7.24-7.35 (m, 3H), 6.51 (s, 1H), 6.17 (bs, 2H), 3.56-3.60 (m, 1H), 1.31 (s, 9H), 1.02-1.07 (m, 4H). LCMS: 450.2 [M+H].

Example 5

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.080 g, 0.282 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.088 g, 0.282 mmol) as starting materials and was obtained as a white solid (0.031 g, 22% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.59 (bs, 1H), 8.84 (bs, 1H), 8.11-8.17 (m, 2H), 7.26-7.37 (m, 3H), 6.20 (s, 1H), 6.16 (bs, 2H), 3.55-3.61 (m, 1H), 1.45-1.49 (m, 2H), 1.38-1.43 (m, 2H), 1.03-1.08 (m, 4H). LCMS: 502.1 [M+H].

Example 6

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.060 g, 0.180 mmol) and phenyl (5-cyclopropylisoxazol-3-yl)carbamate (E5, 0.044 g, 0.282 mmol) as starting materials and was obtained as an off-white solid (8.4 mg, 9% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.20-8.22 (m, 2H), 7.28-7.33 (m, 2H), 7.22 (s, 1H), 6.35 (s, 1H), 3.49-3.55 (m, 1H), 2.08-2.12 (m, 1H), 1.08-1.16 (m, 6H), 0.97-0.99 (m, 2H). LCMS: 434.2 [M+H].

Example 7

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-methylisoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.100 g, 0.35 mmol) and phenyl (3-methylisoxazol-5-yl)carbamate (E3, 0.077 g, 0.35 mmol) as starting materials and was obtained as an off-white solid (0.024 g, 17% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.35 (bs, 1H), 8.84 (bs, 1H), 8.10-8.18 (m, 2H), 7.25-7.36 (m, 3H), 6.13 (bs, 2H), 5.99 (s, 1H), 3.55-3.61 (m, 1H), 2.18 (s, 3H), 1.00-1.08 (m, 4H). LCMS: 408.1 [M+H].

Example 8

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(5-methylisoxazol-3-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.127 g, 0.44 mmol) and phenyl (5-methylisoxazol-3-yl)carbamate (E4, 0.097 g, 0.44 mmol) as starting materials and was obtained as a white solid (0.033 g, 18% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=9.88 (bs, 1H), 8.96 (bs, 1H), 8.15-8.19 (m, 2H), 7.24-7.35 (m, 3H), 6.54 (s, 1H), 6.15 (bs, 2H), 3.55-3.61 (m, 1H), 2.38 (s, 3H), 1.00-1.07 (m, 4H). LCMS: 408.2 [M+H].

Example 9

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(2-fluoropropan-2-yl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.076 g, 0.26 mmol) and phenyl (3-(2-fluoropropan-2-yl)isoxazol-5-yl)carbamate (E9, 0.070 g, 0.26 mmol) as starting materials and was obtained as a white solid (0.018 g, 14% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=8.99 (bs, 1H), 8.17 (s, 1H), 8.09-8.14 (m, 1H), 7.25-7.36 (m, 3H), 6.18 (s, 1H), 6.09 (bs, 2H), 3.57-3.61 (m, 1H), 1.71 (s, 3H), 1.66 (s, 3H), 1.02-1.05 (m, 4H). LCMS: 454.2 [M+H].

Example 10

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.10 g, 0.35 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-yl)carbamate (E7, 0.11 g, 0.35 mmol) as starting materials and was obtained as an off-white solid (0.010 g, 6% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=9.99 (bs, 1H), 8.86 (bs, 1H), 8.14-8.18 (m, 2H), 7.24-7.36 (m, 3H), 6.90 (s, 1H), 6.15 (bs, 2H), 3.56-3.61 (m, 1H), 1.48-1.57 (m, 4H), 1.02-1.07 (m, 4H). LCMS: 502.2 [M+H].

Example 11

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-3-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-2-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D5, 0.070 g, 0.24 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.082 g, 0.24 mmol) as starting materials and was obtained as an off-white solid (0.011 g, 9% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.56 (bs, 1H), 9.29 (bs, 1H), 8.16 (s, 1H), 7.58-7.62 (m, 1H), 7.27-7.36 (m, 2H), 7.22 (s, 1H), 6.20 (s, 1H), 6.00 (bs, 2H), 3.55-3.60 (m, 1H), 1.38-1.48 (m, 4H), 1.02-1.06 (m, 4H). LCMS: 502.2 [M+H].

Example 12

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1,1,1-trifluoro-2-methylpropan-2-yl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-2-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.050 g, 0.176 mmol) and phenyl (3-(1,1,1-trifluoro-2-methylpropan-2-yl)isoxazol-5-yl)carbamate (E8, 0.055 g, 0.176 mmol) as starting materials and was obtained as an off-white solid (9.9 mg, 11% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.59 (bs, 1H), 8.83 (bs, 1H), 8.18 (s, 1H), 8.12-8.17 (m, 1H), 7.25-7.36 (m, 3H), 6.22 (s, 1H), 6.17 (bs, 2H), 3.55-3.61 (m, 1H), 1.52 (s, 6H), 1.02-1.07 (m, 4H). LCMS: 504.2 [M+H].

Example 13

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-3-(3-(tert-butyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-aminophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D4, 0.080 g, 0.30 mmol) and phenyl (3-(tert-butyl)isoxazol-5-yl)carbamate (E1, 0.078 g, 0.30 mmol) as starting materials and was obtained as an off-white solid (0.036 g, 28% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.13 (bs, 1H), 8.93 (bs, 1H), 8.16 (s, 1H), 7.56 (d, J=8.4 Hz, 2H), 7.40 (d, J=8.8 Hz, 2H), 7.22 (s, 1H), 6.08 (s, 1H), 6.00 (bs, 2H), 3.56-3.59 (m, 1H), 1.27 (s, 9H), 1.05-1.07 (m, 4H). LCMS: 430.2 [MH].

Example 14

1-(4-(4-amino-7-(2-hydroxy-2-methylpropyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 1-(4-amino-5-(4-amino-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol (D3, 0.086 g, 0.273 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.085 g, 0.273 mmol) as starting materials and was obtained as a pale brown solid (0.011 g, 8% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.92 (bs, 1H), 9.06 (bs, 1H), 8.11-8.15 (m, 2H), 7.26-7.35 (m, 3H), 6.18 (s, 1H), 6.14 (bs, 2H), 4.86 (bs, 1H), 4.11 (bs, 2H), 1.37-1.46 (m, 4H), 1.06-1.08 (m, 6H). LCMS: 534.1 [M+H].

Example 15

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)cyclohex-3-en-1-yl)-3-(3-(tert-butyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-aminocyclohex-1-en-1-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D7, 0.120 g, 0.446 mmol) and phenyl (3-(tert-butyl)isoxazol-5-yl)carbamate (E1, 0.116 g, 0.446 mmol) as starting materials and was obtained as an off-white solid (0.013 g, 7% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=9.83 (bs, 1H), 8.39 (bs, 1H), 7.47 (s, 1H), 6.54 (d, J=7.6 Hz, 1H), 5.93 (s, 1H), 5.67 (bs, 1H), 3.88-3.90 (m, 2H), 3.61-3.67 (m, 2H), 2.08-2.14 (m, 1H), 1.92-1.95 (m, 1H), 1.69-1.73 (m, 1H), 1.24 (s, 9H), 1.05-1.08 (m, 4H). LCMS: 436.2 [M+H].

Example 16

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)cyclohexyl)-3-(3-(tert-butyl)isoxazol-5-yl)urea

Platinum oxide (0.016 g, 0.069 mmol) was added to a solution of 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)cyclohex-3-en-1-yl)-3-(3-(tert-butyl)isoxazol-5-yl)urea (Example 15, 0.100 g, 0.230 mmol) in EtOAc (5 mL) and the resulting suspension was stirred at room temperature under _H2 for 12 h. After the reaction was complete (as determined by LCMS), the reaction mixture was filtered through a pad of celite, which was then washed with EtOAc (2×5 mL). The combined filtrates were concentrated under reduced pressure to give the crude material, which was purified by preparative HPLC (mass detection, eluting with a gradient of ammonium acetate in water and ACN) to give the title product as an off-white solid (2.0 mg, 2% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ = 8.12 (s, 1H), 6.92 (s, 1H), 6.02 (s, 1H), 3.50-3.51 (m, 1H), 2.87-2.90 (m, 1H), 2.13-2.18 (m, 6H), 1.51-1.57 (m, 3H), 1.33 (s, 9H), 0.90-1.20 (m, 4H). LCMS: 436.2 [MH].

Example 17

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-methylcyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.050 g, 0.176 mmol) and phenyl (3-(1-methylcyclopropyl)isoxazol-5-yl)carbamate (E10, 0.046 g, 0.176 mmol) as starting materials and was obtained as an off-white solid (0.013 g, 16% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.40 (bs, 1H), 8.84 (bs, 1H), 8.11-8.17 (m, 2H), 7.24-7.36 (m, 3H), 6.16 (bs, 2H), 5.84 (s, 1H), 3.55-3.61 (m, 1H), 1.38 (s, 3H), 1.02-1.07 (m, 4H), 0.94-0.96 (m, 2H), 0.83-0.84 (m, 2H). LCMS: 448.2 [M+H].

Example 18

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(4-(tert-butyl)thiazol-2-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (E11, 0.040 g, 0.099 mmol) and 4-(tert-butyl)thiazol-2-amine (0.015 g, 0.099 mmol) as starting materials and was obtained as an off-white solid (7.0 mg, 15% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.99 (bs, 1H), 9.29 (bs, 1H), 8.46 (s, 1H), 8.26-8.30 (m, 1H), 7.67 (s, 1H), 7.38-7.41 (m, 1H), 7.27-7.30 (m, 1H), 6.70 (s, 1H), 3.71 (bs, 1H), 1.27 (s, 9H), 1.10-1.10 (m, 4H). LCMS: 466.0 [M+H].

Example 19

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(5-(tert-butyl)-1,3,4-thiadiazol-2-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.050 g, 0.176 mmol) and phenyl (5-(tert-butyl)-1,3,4-thiadiazol-2-yl) carbamate (E12, 0.049 g, 0.176 mmol) as starting materials and was obtained as an off-white solid (2.0 mg, 2% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.19-8.23 (m, 2H), 7.30-7.35 (m, 2H), 7.23 (s, 1H), 3.50-3.54 (m, 1H), 1.49 (s, 9H), 1.08-1.17 (m, 4H). LCMS: 466.9 [M+H].

Example 20

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(tert-butyl)isothiazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (E11, 0.040 g, 0.099 mmol) and 4-(tert-butyl)thiazol-2-amine (0.015 g, 0.099 mmol) as starting materials and was obtained as an off-white solid (2.0 mg, 4% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.21 (s, 1H), 8.11-8.17 (m, 1H), 7.30-7.34 (m, 2H), 7.23 (s, 1H), 6.78 (s, 1H), 3.50-3.54 (m, 1H), 1.35 (s, 9H), 1.08-1.30 (m, 4H). LCMS: 466.0 [M+H].

Example 21

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(tert-butyl)-1,2,4-thiadiazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.050 g, 0.176 mmol) and phenyl (3-(tert-butyl)-1,2,4-thiadiazol-5-yl) carbamate (E13, 0.049 g, 0.176 mmol) as starting materials and was obtained as an off-white solid (8.0 mg, 10% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=11.54 (bs, 1H), 9.00 (bs, 1H), 8.09-8.18 (m, 2H), 7.28-7.39 (m, 3H), 6.19 (bs, 2H), 3.57-3.61 (m, 1H), 1.34 (s, 9H), 1.03-1.05 (m, 4H). LCMS: 467.0 [M+H].

Example 22

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(1-(tert-butyl)-1H-1,2,4-triazol-3-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (E11, 0.040 g, 0.099 mmol) and 1-(tert-butyl)-1H-1,2,4-triazol-3-amine (0.014 g, 0.099 mmol) as starting materials and was obtained as an off-white solid (3.0 mg, 6% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.70 (bs, 1H), 10.16 (bs, 1H), 8.54 (s, 1H), 8.31-8.35 (m, 1H), 8.16 (s, 1H), 7.25-7.37 (m, 3H), 6.17 (bs, 2H), 3.56-3.59 (m, 1H), 1.57 (s, 9H), 1.02-1.05 (m, 4H). LCMS: 450.0 [M+H].

Example 23

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(5-(tert-butyl)-1,3,4-oxadiazol-2-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using phenyl (4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)carbamate (E11, 0.040 g, 0.099 mmol) and 5-(tert-butyl)-1,3,4-oxadiazol-2-amine (0.014 g, 0.099 mmol) as starting materials and was obtained as an off-white solid (2.0 mg, 4% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.35 (s, 1H), 8.26-8.30 (m, 1H), 7.48 (s, 1H), 7.31-7.40 (m, 2H), 3.69-3.71 (m, 1H), 1.45 (s, 9H), 1.17-1.22 (m, 4H). LCMS: 451.0 [M+H].

Example 24

1-(4-(4-amino-7-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-(pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D8, 0.100 g, 0.312 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.107 g, 0.343 mmol) as starting materials and was obtained as a white solid (0.023 g, 14% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.69 (bs, 1H), 9.13 (bs, 1H), 8.97 (bs, 1H), 8.62 (d, J=4.8 Hz, 1H), 8.31-8.36 (m, 2H), 8.20-8.24 (m, 1H), 7.95 (s, 1H), 7.62-7.65 (m, 1H), 7.38-7.50 (m, 2H), 6.76 (bs, 2H), 6.21 (s, 1H), 1.39-1.47 (m, 4H). LCMS: 538.8 [M+H].

Example 25

1-(4-(4-amino-7-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D9, 0.080 g, 0.250 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.086 g, 0.275 mmol) as starting materials and was obtained as an off-white solid (0.046 g, 33% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.68 (bs, 1H), 9.00 (bs, 1H), 8.89 (d, J=6.4 Hz, 2H), 8.58 (d, J=6.4 Hz, 2H), 8.43 (s, 1H), 8.21-8.27 (m, 2H), 7.49-7.52 (m, 1H), 7.39-7.42 (m, 1H), 6.90 (bs, 2H), 6.22 (s, 1H), 1.39-1.49 (m, 4H). LCMS: 538.9 [M+H].

Example 26

1-(4-(4-amino-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D10, 0.020 g, 0.059 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.018 g, 0.059 mmol) as starting materials and was obtained as a white solid (2.4 mg, 7% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.68 (bs, 1H), 8.96 (bs, 1H), 8.32 (s, 1H), 8.16-8.20 (m, 1H), 7.55 (s, 1H), 7.38-7.41 (m, 1H), 7.29-7.31 (m, 1H), 6.98 (bs, 2H), 6.21 (s, 1H), 4.85-4.91 (m, 1H), 3.59-3.62 (m, 4H), 2.85 (s, 3H), 2.21-2.34 (m, 4H), 1.38-1.49 (m, 4H). LCMS: 559.2 [M+H].

Example 27

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(3-methyloxetan-3-yl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.050 g, 0.176 mmol) and phenyl (3-(3-methyloxetan-3-yl)isoxazol-5-yl)carbamate (E14, 0.048 g, 0.176 mmol) as starting materials and was obtained as a white solid (3.4 mg, 4% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.56 (s, 1H), 8.94 (bs, 1H), 8.42 (s, 1H), 8.19 (t, J=8.4 Hz, 1H), 7.57 (bs, 3H), 7.38 (d, J=10.8 Hz, 1H), 7.28 (d, J=8.4 Hz, 1H), 6.22 (s, 1H), 4.77 (d, J=5.6 Hz, 2H), 4.52 (d, J=5.6 Hz, 2H), 3.68-3.69 (m, 1H), 1.63 (s, 3H), 1.09-1.10 (m, 4H). LCMS: 464.1 [M+H].

Example 28

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(trifluoromethyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.050 g, 0.176 mmol) and phenyl (3-(trifluoromethyl)isoxazol-5-yl)carbamate (E15, 0.048 g, 0.176 mmol) as starting materials and was obtained as a white solid (2.9 mg, 4% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=11.03 (bs, 1H), 8.97 (bs, 1H), 8.19 (s, 1H), 8.10 (t, J=8.4 Hz, 1H), 7.27-7.38 (m, 3H), 6.54 (s, 1H), 6.30 (bs, 2H), 3.56-3.60 (m, 1H), 1.03-1.05 (m, 4H). LCMS: 461.9 [M+H].

Example 29

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-hydroxy-2-methylpropan-2-yl)isoxazol-5-yl)urea

Step 1: Synthesis of 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-((tert-butyldimethylsilyl)oxy)-2-methylpropan-2-yl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.131 g, 0.461 mmol) and phenyl (3-(1-((tert-butyldimethylsilyl)oxy)-2-methylpropan-2-yl)isoxazol-5-yl)carbamate (E16, 0.180 g, 0.461 mmol) as an off-white solid (0.013 g, 5% yield). LCMS: 580.0 [M+H].

Step 2: Synthesis of 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-hydroxy-2-methylpropan-2-yl)isoxazol-5-yl)urea

TBAF (1 M in THF, 0.067 mL) was added to a solution of 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-((tert-butyldimethylsilyl)oxy)-2-methylpropan-2-yl)isoxazol-5-yl)urea (0.013 g, 0.022 mmol) in THF (2 mL) at 0 °C and the resulting solution was stirred at 25 °C for 4 h. After completion of the reaction (as determined by TLC), the reaction mixture was concentrated under reduced pressure to give the crude material, which was purified by preparative HPLC to afford the title compound as a white solid (TFA salt, 2.2 mg, 21% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.36 (s, 1H), 8.23 (t, J=8.4 Hz, 1H), 7.49 (s, 1H), 7.30-7.38 (m, 2H), 6.20 (s, 1H), 3.69-3.72 (m, 1H), 3.61 (s, 2H), 1.32 (s, 6H), 1.18-1.24 (m, 4H). LCMS: 466.2 [M+H].

Example 30

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(sec-butyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.055 g, 0.194 mmol) and phenyl (3-(sec-butyl)isoxazol-5-yl)carbamate (E17, 0.051 g, 0.194 mmol) as starting materials and was obtained as a white solid (0.012 g, 14% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.39 (bs, 1H), 8.84 (bs, 1H), 8.12-8.14 (m, 2H), 7.25-7.36 (m, 3H), 6.16 (bs, 2H), 6.02 (s, 1H), 3.56-3.59 (m, 1H), 2.68-2.73 (m, 1H), 1.56-1.60 (m, 2H), 1.18-1.20 (m, 3H), 1.01-1.10 (m, 4H), 0.81-0.89 (m, 3H). LCMS: 450.0 [M+H].

Example 31

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(pentan-3-yl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.050 g, 0.176 mmol) and phenyl (3-(pentan-3-yl)isoxazol-5-yl)carbamate (E18, 0.048 g, 0.176 mmol) as starting materials and was obtained as an off-white solid (0.016 g, 19% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.65 (bs, 1H), 9.06 (bs, 1H), 8.37 (s, 1H), 8.18 (t, J=8.4 Hz, 1H), 7.51 (s, 1H), 7.35-7.38 (m, 1H), 7.26-7.28 (m, 3H), 5.98 (s, 1H), 3.65-3.68 (m, 1H), 1.49-1.68 (m, 4H), 1.07-1.09 (m, 4H), 0.79-0.82 (m, 6H). LCMS: 464.0 [M+H].

Example 32

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-isopropyloxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.056 g, 0.198 mmol) and phenyl (3-isopropyloxazol-5-yl)carbamate (E19, 0.049 g, 0.198 mmol) as starting materials and was obtained as an off-white solid (0.018 g, 21% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.34 (bs, 1H), 8.81 (bs, 1H), 8.12-8.16 (m, 2H), 7.25-7.36 (m, 3H), 6.15 (bs, 2H), 6.04 (s, 1H), 3.55-3.59 (m, 1H), 2.89-2.96 (m, 1H), 1.21-1.25 (m, 6H), 1.07-1.09 (m, 4H). LCMS: 436.0 [M+H].

Example 33

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-ethylisoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.061 g, 0.215 mmol) and phenyl (3-ethylisoxazol-5-yl)carbamate (E20, 0.050 g, 0.215 mmol) as starting materials and was obtained as an off-white solid (0.020 g, 22% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.31 (bs, 1H), 8.81 (bs, 1H), 8.12-8.17 (m, 2H), 7.25-7.36 (m, 3H), 6.15 (bs, 2H), 6.03 (s, 1H), 3.55-3.60 (m, 1H), 2.54-2.60 (m, 2H), 1.05-1.21 (m, 3H), 1.03-1.04 (m, 4H). LCMS: 422.0 [M+H].

Example 34

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-methylcyclobutyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.075 g, 0.265 mmol) and phenyl (3-(1-methylcyclobutyl)isoxazol-5-yl)carbamate (E21, 0.072 g, 0.265 mmol) as starting materials and was obtained as a white solid (6.8 mg, 5% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.42 (bs, 1H), 8.90 (bs, 1H), 8.42 (s, 1H), 8.18-8.22 (m, 1H), 7.58 (s, 1H), 7.37-7.40 (m, 1H), 7.27-7.29 (m, 1H), 6.06 (s, 1H), 3.67-3.71 (m, 1H), 2.34-2.38 (m, 2H), 1.85-2.08 (m, 4H), 1.42 (s, 3H), 1.07-1.11 (m, 4H). LCMS: 462.2 [M+H].

Example 35

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(2-cyanopropan-2-yl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.070 g, 0.247 mmol) and phenyl (3-(2-cyanopropan-2-yl)isoxazol-5-yl)carbamate (E22, 0.067 g, 0.247 mmol) as starting materials and was obtained as a white solid (4.1 mg, 3% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.33 (s, 1H), 8.20-8.24 (m, 1H), 7.43 (s, 1H), 7.31-7.38 (m, 2H), 6.33 (s, 1H), 3.50-3.66 (m, 1H), 1.77 (s, 6H), 1.15-1.21 (m, 4H). LCMS: 461.1 [M+H].

Example 36

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(hydroxymethyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.250 g, 0.882 mmol) and phenyl (3-(((tert-butyldimethylsilyl)oxy)methyl)isoxazol-5-yl)carbamate (E23, 0.308 g, 0.882 mmol) as starting materials and was obtained as an off-white solid (0.029 g, 8% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.43 (bs, 1H), 8.92 (bs, 1H), 8.40 (s, 1H), 8.18-8.22 (m, 1H), 7.56 (s, 1H), 7.37-7.40 (m, 1H), 7.27-7.29 (m, 1H), 6.12 (s, 1H), 4.43 (s, 2H), 3.66-3.71 (m, 1H), 1.08-1.11 (m, 4H). LCMS: 422.0 [MH].

Note: When cleaned up by gradient elution with 10 mM ammonium acetate in water and ACN, cleavage of the TBDMS group was observed.

Example 37

1-(5-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pyridin-2-yl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(6-aminopyridin-3-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D6, 0.300 g, 1.127 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.352 g, 1.127 mmol) as starting materials and was obtained as an off-white solid (0.018 g, 3% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=11.77 (bs, 1H), 9.85 (bs, 1H), 8.39-8.40 (m, 1H), 8.18 (s, 1H), 7.86-7.88 (m, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.34 (s, 1H), 6.26 (s, 1H), 6.23 (bs, 2H), 3.58-3.61 (m, 1H), 1.40-1.49 (m, 4H), 1.03-1.06 (m, 4H). LCMS: 485.0 [M+H].

Example 38

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylphenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-methylphenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D11, 0.270 g, 0.967 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.302 g, 0.967 mmol) as starting materials and was obtained as an off-white solid (0.080 g, 16% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.63 (bs, 1H), 8.29 (bs, 1H), 8.16 (s, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.22-7.32 (m, 3H), 6.17 (s, 1H), 6.08 (bs, 2H), 3.55-3.60 (m, 1H), 2.29 (s, 3H), 1.37-1.48 (m, 4H), 1.00-1.05 (m, 4H). LCMS: 498.1 [M+H].

Example 39

1-(4-(4-amino-7-(3-hydroxycyclobutyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

Step 1: Synthesis of 1-(4-(4-amino-7-(3-(benzyloxy)cyclobutyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(4-amino-3-fluorophenyl)-7-(3-(benzyloxy)cyclobutyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D12, 0.033 g, 0.082 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.028 g, 0.090 mmol) as a yellow gum (0.038 g, 29% yield). LCMS: 622.3 [M+H].

Step 2: Synthesis of 1-(4-(4-amino-7-(3-hydroxycyclobutyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

Boron trichloride (1 M in DCM, 0.901 mL, 0.901 mmol) was added dropwise to a solution of 1-(4-(4-amino-7-(3-(benzyloxy)cyclobutyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea (0.070 g, 0.113 mmol) in DCM (5 mL) at -60 °C and the resulting mixture was stirred at 0 °C for 3 h. After completion of the reaction (as determined by TLC and LCMS), the reaction mixture was cooled to -70 °C, neutralized with NH ₄ OH (25% in water) and extracted with DCM (2×10 mL). The combined organic extracts were dried over _Na2SO4 , _filtered , and concentrated under reduced pressure to give a crude material, which was purified by preparative HPLC (eluting with a gradient of 1% TFA in water and ACN) to give the title product as a white solid (0.012 g, 20% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.33 (s, 1H), 8.22-8.26 (m, 1H), 7.77 (s, 1H), 7.34-7.42 (m, 2H), 6.33 (s, 1H), 5.58-5.62 (m, 1H), 4.64-4.66 (m, 1H), 2.85-2.92 (m, 2H), 2.58-2.64 (m, 2H), 1.39-1.49 (m, 4H). LCMS: 531.8 [M+H].

Example 40

1-(6-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-4-methylpyridin-3-yl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(5-amino-4-methylpyridin-2-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D13, 0.050 g, 0.102 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.032 g, 0.102 mmol) as starting materials and was obtained as an off-white solid (8 mg, 16% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=9.81 (bs, 1H), 8.76 (s, 1H), 8.51 (bs, 1H), 8.08 (s, 1H), 7.93-7.94 (m, 2H), 7.12 (bs, 2H), 6.17 (s, 1H), 3.53-3.56 (m, 1H), 2.30 (s, 3H), 1.37-1.46 (m, 4H), 1.05-1.08 (m, 4H). LCMS: 499.2 [M+H].

Example 41

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2,6-difluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(4-amino-3,5-difluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D15, 0.080 g, 0.266 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.083 g, 0.266 mmol) as starting materials and was obtained as an off-white solid (8 mg, 5.6% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.78 (bs, 1H), 8.71 (bs, 1H), 8.42 (bs, 1H), 7.67 (s, 1H), 7.56 (bs, 2H), 7.23-7.29 (m, 2H), 6.14 (s, 1H), 3.67-3.76 (m, 1H), 1.37-1.47 (m, 4H), 1.09-1.12 (m, 4H). LCMS: 519.7 [M+H].

Example 42

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2,5-difluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(4-amino-2,5-difluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D16, 0.100 g, 0.332 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.104 g, 0.332 mmol) as starting materials and was obtained as an off-white solid (8 mg, 5% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.63 (bs, 1H), 9.01 (bs, 1H), 8.16 (s, 1H), 8.04-8.09 (m, 1H), 7.28-7.34 (m, 2H), 6.21 (s, 1H), 6.16 (bs, 2H), 3.55-3.61 (m, 1H), 1.36-1.49 (m, 4H), 1.00-1.04 (m, 4H). LCMS: 520.1 [M+H].

Example 43

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-3,5-difluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(4-amino-2,6-difluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D17, 0.0724 g, 0.240 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.075 g, 0.240 mmol) as starting materials and was obtained as an off-white solid (3 mg, 2% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=8.14 (s, 1H), 7.39-7.42 (m, 2H), 7.24 (s, 1H), 6.15 (s, 1H), 6.02 (bs, 2H), 3.58-3.59 (m, 1H), 1.36-1.44 (m, 4H), 1.03-1.05 (m, 4H). LCMS: 520.2 [M+H].

Example 44

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-3,5-difluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

Step 1: Synthesis of 1-(5-bromo-3-fluoropyridin-2-yl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (Method B) using 5-bromo-3-fluoropyridin-2-amine (0.040 g, 0.209 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.065 g, 0.209 mmol) as starting materials and was obtained as a pale yellow solid (0.020 g, 23% yield). LCMS: 410.9 [M+H].

Step 2: Synthesis of 1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-3,5-difluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared by a procedure similar to that described for intermediate D8 using 1-(5-bromo-3-fluoropyridin-2-yl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea (0.020 g, 0.049 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 PCT Publication No. WO 2018/015879, 0.024 g, 0.049 mmol) as starting materials and was obtained as an off-white solid (2 mg, 7% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.32-8.33 (m, 2H), 7.81-7.84 (m, 1H), 7.50 (s, 1H), 6.42 (s, 1H), 3.60-3.68 (m, 1H), 1.41-1.50 (m, 4H), 1.13-1.23 (m, 4H). LCMS: 503.1 [M+H]. Note: elimination of Boc groups was observed during the reaction.

Example 45

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclobutyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.080 g, 0.282 mmol) and phenyl (3-(1-(trifluoromethyl)cyclobutyl)isoxazol-5-yl)carbamate (E24, 0.092 g, 0.282 mmol) as starting materials and was obtained as an off-white solid (0.035 g, 22% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.66 (bs, 1H), 8.90 (bs, 1H), 8.21 (s, 1H), 8.12-8.16 (m, 1H), 7.26-7.36 (m, 3H), 6.41 (bs, 2H), 6.14 (s, 1H), 3.54-3.56 (m, 1H), 2.57-2.68 (m, 4H), 2.03-2.05 (m, 2H), 1.04-1.06 (m, 4H). LCMS: 515.9 [M+H].

Example 46

1-(5-(4-amino-7-(2-hydroxy-2-methylpropyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pyridin-2-yl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 1-(4-amino-5-(6-aminopyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol (D18, 0.100 g, 0.335 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.105 g, 0.335 mmol) as starting materials and was obtained as an off-white solid (0.010 g, 6% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=11.74 (bs, 1H), 9.91 (bs, 1H), 8.41-8.43 (m, 2H), 7.89-7.92 (m, 1H), 7.80 (bs, 2H), 7.68-7.71 (m, 1H), 7.59 (s, 1H), 6.26 (s, 1H), 4.20 (s, 2H), 1.38-1.50 (m, 4H), 1.11-1.12 (m, 6H). LCMS: 516.9 [M+H].

Example 47

1-(4-(4-amino-7-(2-hydroxy-2-methylpropyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 1-(4-amino-5-(4-amino-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylpropan-2-ol (D3, 0.120 g, 0.381 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-yl)carbamate (E7, 0.119 g, 0.381 mmol) as starting materials and was obtained as an off-white solid (0.028 g, 13% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.04 (bs, 1H), 8.94 (bs, 1H), 8.41 (s, 1H), 8.23-8.27 (m, 1H), 7.55 (s, 1H), 7.36-7.40 (m, 1H), 7.27-7.30 (m, 1H), 6.91 (s, 1H), 4.18 (s, 2H), 1.51-1.58 (m, 4H), 1.07-1.11 (m, 6H). LCMS: 534.2 [M+H].

Example 48

1-(5-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pyrimidin-2-yl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(2-aminopyrimidin-5-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D19, 0.080 g, 0.299 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.093 g, 0.299 mmol) as starting materials and was obtained as an off-white solid (4 mg, 3% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.76 (bs, 1H), 9.04 (bs, 1H), 8.65 (bs, 1H), 8.34 (bs, 2H), 7.61 (s, 1H), 6.66 (bs, 2H), 6.10 (s, 1H), 3.66-3.71 (m, 1H), 1.37-1.45 (m, 4H), 1.11-1.11 (m, 4H). LCMS: 485.9 [M+H].

Example 49

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-(hydroxymethyl)phenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using (2-amino-5-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)methanol (D20, 0.020 g, 0.068 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.021 g, 0.068 mmol) as starting materials and was obtained as an off-white solid (3 mg, 8% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.96 (bs, 1H), 8.56 (bs, 1H), 8.17 (s, 1H), 7.91-7.93 (m, 1H), 7.34-7.42 (m, 2H), 7.21 (s, 1H), 6.04-6.17 (m, 3H), 5.49 (bs, 1H), 4.57 (bs, 2H), 3.56-3.61 (m, 1H), 1.37-1.47 (m, 4H), 1.00-1.06 (m, 4H). LCMS: 514.1 [M+H].

Example 50

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-cyanophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 2-amino-5-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)benzonitrile (D21, 0.130 g, 0.287 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.089 g, 0.287 mmol) as starting materials and was obtained as an off-white solid (5 mg, 3% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=8.62 (s, 1H), 7.43-7.52 (m, 3H), 6.86 (s, 1H), 6.05 (s, 1H), 3.67-3.70 (m, 1H), 1.38-1.46 (m, 4H), 1.08-1.10 (m, 4H). LCMS: 509.2 [M+H].

Example 51

1-(4-(4-amino-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(4-amino-3-fluorophenyl)-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D22, 0.075 g, 0.249 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.078 g, 0.249 mmol) as starting materials and was obtained as an off-white solid (0.037 g, 28% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.63 (bs, 1H), 8.88 (bs, 1H), 8.13-8.18 (m, 2H), 7.42 (s, 1H), 7.26-7.36 (m, 2H), 6.35 (bs, 2H), 6.20 (s, 1H), 4.34 (t, J=5.6 Hz, 2H), 3.72 (t, J=5.2 Hz, 2H), 3.26 (s, 3H), 1.38-1.49 (m, 4H). LCMS: 520.2 [M+H].

Example 52

1-(4-(4-amino-7-(2-hydroxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 2-(4-amino-5-(4-amino-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-ol (D23, 0.080 g, 0.278 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.087 g, 0.278 mmol) as starting materials and was obtained as an off-white solid (0.011 g, 7% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.53 (bs, 1H), 8.78 (bs, 1H), 8.05-8.09 (m, 2H), 7.32 (s, 1H), 7.19-7.28 (m, 2H), 6.18 (bs, 2H), 6.13 (s, 1H), 4.90 (t, J=5.2 Hz, 1H), 4.14 (t, J=6.0 Hz, 2H), 3.66-3.69 (m, 2H), 1.31-1.41 (m, 4H). LCMS: 506.2 [M+H].

Example 53

1-(4-(4-amino-7-cyclobutyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(4-amino-3-fluorophenyl)-7-cyclobutyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D24, 0.150 g, 0.504 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.158 g, 0.504 mmol) as starting materials and was obtained as an off-white solid (0.111 g, 38% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.67 (bs, 1H), 8.96 (bs, 1H), 8.38 (s, 1H), 8.18-8.22 (m, 1H), 7.93 (s, 1H), 7.43 (bs, 2H), 7.41 (d, J=1.6 Hz, 1H), 7.32 (d, J=8.4 Hz, 1H), 6.21 (s, 1H), 5.23-5.27 (m, 1H), 2.51-2.68 (m, 4H), 1.84-1.89 (m, 2H), 1.38-1.49 (m, 4H). LCMS: 516.1 [M+H].

Example 54

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-chlorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(4-amino-3-chlorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D25, 0.050 g, 0.167 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.052 g, 0.167 mmol) as starting materials and was obtained as an off-white solid (6 mg, 7% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=11.04 (bs, 1H), 8.67 (bs, 1H), 8.40 (s, 1H), 8.23-8.25 (m, 1H), 7.58-7.60 (m, 2H), 7.42-7.44 (m, 1H), 6.21 (s, 1H), 3.67-3.73 (m, 1H), 1.46-2.33 (m, 2H), 1.37-1.38 (m, 2H), 1.11-1.13 (m, 4H). LCMS: 518.2 [M+H].

Example 55

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methoxyphenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(4-amino-3-methoxyphenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D26, 0.020 g, 0.068 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.021 g, 0.068 mmol) as starting materials and was obtained as an off-white solid (7 mg, 19% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.96 (bs, 1H), 8.65 (bs, 1H), 8.23 (s, 1H), 8.17 (d, J=8.4 Hz, 1H), 7.30 (s, 1H), 7.12-7.17 (m, 1H), 7.01-7.04 (m, 1H), 6.47 (bs, 2H), 6.19 (s, 1H), 3.94 (s, 3H), 3.60-3.62 (m, 1H), 1.45-1.48 (m, 2H), 1.36-1.38 (m, 2H), 1.04-1.07 (m, 4H). LCMS: 513.9 [M+H].

Example 56

1-(4-(4-amino-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(4-amino-3-fluorophenyl)-7-(1-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D27, 0.040 g, 0.123 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.036 g, 0.115 mmol) as starting materials and was obtained as a white solid (0.015 mg, 24% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.82 (bs, 1H), 9.07 (bs, 1H), 8.39 (s, 1H), 8.19-8.23 (m, 1H), 7.84 (s, 1H), 7.14-7.40 (m, 2H), 6.21 (s, 1H), 5.55-5.66 (m, 1H), 3.90-4.09 (m, 2H), 2.95 (bs, 4H), 2.08 (s, 3H), 1.38-1.49 (m, 4H). LCMS: 545.3 [M+H].

Example 57

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(5-(1-(trifluoromethyl)cyclobutyl)isoxazol-3-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3-fluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D1, 0.160 g, 0.565 mmol) and phenyl (5-(1-(trifluoromethyl)cyclobutyl)isoxazol-3-yl)carbamate (E25, 0.184 g, 0.565 mmol) as starting materials and was obtained as a white solid (0.081 mg, 28% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.02 (bs, 1H), 8.87 (bs, 1H), 8.15-8.20 (m, 2H), 7.25-7.36 (m, 3H), 6.96 (s, 1H), 6.16 (bs, 2H), 3.55-3.61 (m, 1H), 2.59-2.68 (m, 4H), 2.03-2.11 (m, 2H), 1.02-1.11 (m, 4H). LCMS: 516.2 [M+H].

Example 58

1-(4-(4-amino-7-cyclobutyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(4-amino-3-fluorophenyl)-7-cyclobutyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D24, 0.204 g, 0.686 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-yl)carbamate (E7, 0.214 g, 0.686 mmol) as starting materials and was obtained as an off-white solid (0.096 g, 27% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.01 (bs, 1H), 8.87 (bs, 1H), 8.14-8.20 (m, 2H), 7.66 (s, 1H), 7.28-7.39 (m, 2H), 6.91 (s, 1H), 6.18 (bs, 2H), 5.18-5.23 (m, 1H), 2.39-2.41 (m, 4H), 1.81-1.90 (m, 2H), 1.54-1.55 (m, 4H). LCMS: 516.2 [M+H].

Example 59

1-(6-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)pyridin-3-yl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 5-(5-aminopyridin-2-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D14, 0.030 g, 0.045 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.014 g, 0.045 mmol) as starting materials and was obtained as an off-white solid (5 mg, 23% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=10.67 (bs, 1H), 9.68 (bs, 1H), 9.12 (s, 1H), 8.59-8.60 (m, 1H), 8.09 (s, 1H), 7.92-8.00 (m, 2H), 7.17 (bs, 2H), 6.20 (s, 1H), 3.58-3.63 (m, 1H), 1.36-1.48 (m, 4H), 1.06-1.10 (m, 4H). LCMS: 485.2 [M+H].

Example 60

1-(4-(4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2,6-difluorophenyl)-3-(5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure A) using 5-(4-amino-3,5-difluorophenyl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (D15, 0.150 g, 0.498 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-yl)carbamate (E7, 0.155 g, 0.498 mmol) as starting materials and was obtained as a white solid (0.015 g, 6% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=11.47 (bs, 1H), 10.17 (bs, 1H), 8.18 (s, 1H), 7.42 (s, 1H), 7.18-7.22 (m, 2H), 6.82 (s, 1H), 6.27 (bs, 2H), 3.51-3.55 (m, 1H), 1.46-1.51 (m, 4H), 1.04-1.05 (m, 4H). LCMS: 520.2 [M+H].

Example 61

1-(4-(4-amino-7-(2-hydroxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 2-(4-amino-5-(4-amino-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-ol (D23, 0.100 g, 0.348 mmol) and phenyl (5-(1-(trifluoromethyl)cyclopropyl)isoxazol-3-yl)carbamate (E7, 0.109 g, 0.348 mmol) as starting materials and was obtained as a white solid (0.035 mg, 20% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.35 (s, 1H), 8.23-8.27 (m, 1H), 7.58 (s, 1H), 7.32-7.39 (m, 2H), 6.81 (s, 1H), 4.47 (t, J=10.8 Hz, 2H), 3.95-3.98 (m, 2H), 1.49-1.59 (m, 4H). LCMS: 506.2 [M+H].

Example 62

1-(4-(4-amino-1-cyclopropyl-1H-pyrrolo[3,2-c]pyridin-3-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

Step 1: Synthesis of 1-cyclopropyl-N-(2,4-dimethoxybenzyl)-3-iodo-1H-pyrrolo[3,2-c]pyridin-4-amine

A mixture of 4-chloro-1-cyclopropyl-3-iodo-1H-pyrrolo[3,2-c]pyridine (B9, 0.300 g, 0.942 mmol) and (2,5-dimethoxyphenyl)methanamine (0.429 mL, 2.83 mmol) in n-BuOH (10 mL) was stirred at 110 °C for 12 h. After the reaction was complete (as determined by LCMS), the reaction mixture was concentrated under reduced pressure to give a crude material, which was purified using Isolera (silica gel 230-400 mesh eluting with 30% EtOAc in petroleum ether) to afford the desired product as a yellow gum (0.10 g, 19% yield). LCMS: 450.0 [M+H].

Step 2: Synthesis of 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-N-(2,4-dimethoxybenzyl)-1H-pyrrolo[3,2-c]pyridin-4-amine

The title compound was prepared by a procedure similar to that described for intermediate D8 using 1-cyclopropyl-N-(2,4-dimethoxybenzyl)-3-iodo-1H-pyrrolo[3,2-c]pyridin-4-amine (0.190 g, 0.423 mmol) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (0.100 g, 0.423 mmol) as a brown gum (0.080 g, 41% yield). LCMS: 433.2 [M+H].

Step 3: Synthesis of 1-(4-(1-cyclopropyl-4-((2,4-dimethoxybenzyl)amino)-1H-pyrrolo[3,2-c]pyridin-3-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

The title compound was prepared according to the general procedure for the preparation of urea (procedure B) using 3-(4-amino-3-fluorophenyl)-1-cyclopropyl-N-(2,4-dimethoxybenzyl)-1H-pyrrolo[3,2-c]pyridin-4-amine (0.080 g, 0.185 mmol) and phenyl (3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)carbamate (E6, 0.058 g, 0.185 mmol) as an off-white solid (0.027 g, 17% yield). LCMS: 651.3 [M+H].

Step 4: Synthesis of 1-(4-(4-amino-1-cyclopropyl-1H-pyrrolo[3,2-c]pyridin-3-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea

Triethylsilane (4.8 mg, 0.041 mmol) and TFA (4.7 mg, 0.041 mmol) were added to a solution of 1-(4-(1-cyclopropyl-4-((2,4-dimethoxybenzyl)amino)-1H-pyrrolo[3,2-c]pyridin-3-yl)-2-fluorophenyl)-3-(3-(1-(trifluoromethyl)cyclopropyl)isoxazol-5-yl)urea (0.027 g, 0.041 mmol) in DCM (2 mL) at 0 °C, and the resulting mixture was stirred at 25 °C for 12 h. After completion of the reaction (as determined by LCMS), the reaction mixture was concentrated under reduced pressure to give a crude material, which was purified by preparative HPLC (eluting with a gradient of 0.1% TFA in water and) and the desired product was obtained as an off-white solid (5 mg, 24% yield). ¹ H NMR (400 MHz, CD ₃ OD) δ=8.21-8.25 (m, 1H), 7.64-7.66 (m, 1H), 7.51 (s, 1H), 7.30-7.38 (m, 3H), 6.33 (s, 1H), 3.59-3.62 (m, 1H), 1.39-1.49 (m, 4H), 1.13-1.26 (m, 4H). LCMS: 501.2 [M+H].

Biological example 1

Biochemical study of compounds

The inhibitory activity of representative compounds on NEK7 and IL-1β release was studied using the methods described above. The results are presented in the following table.

The various embodiments described above can be combined to form other embodiments. All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent publications cited in this specification and/or referenced in the instructions for use, including, but not limited to, U.S. Provisional Patent Application No. 63/022,159, filed May 8, 2020, and U.S. Provisional Patent Application No. 63/170,761, filed April 5, 2021; are hereby incorporated by reference in their entireties. Aspects of the embodiments can be modified, as desired, using the concepts of the various patents, applications, and publications to form additional embodiments.

These and other modifications may be made to the embodiments in light of the above detailed description. The terms commonly used in the following claims should not be construed as limiting the claims to the specific embodiments disclosed in the present description and claims, but should be construed as including all possible embodiments together with the full scope of equivalents of such claims. Accordingly, the claims are not limited to the present invention.

Claims (55)

1. A compound having the following structure (I): or a pharmaceutically acceptable salt thereof, where: A is _C6 aryl, _C6 cycloalkyl, or 6-membered monocyclic heteroaryl containing one to two nitrogen heteroatoms, each of which is optionally substituted with one or two ^R6 ; X stands for N; Y stands for NH; R ¹ means H; R ² means , _C3 - _C4 cycloalkyl or 4- to 6-membered heterocyclyl having one heteroatom selected from nitrogen or oxygen, or 6-membered heteroaryl containing one nitrogen heteroatom, wherein the cycloalkyl or heterocycloalkyl is optionally substituted with one substituent selected from the group consisting of hydroxyl and _C1 - _C6 alkyl; R ³ means H; R ⁴ is a heteroaryl selected from isoxazolyl, 1,2,4-triazolyl, isothiazolyl, 1,2,4-thiadiazolyl and 1,3,4-thiadiazolyl, each of which is optionally substituted with one or more substituents selected from the group consisting of halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ cyanoalkyl, C ₁ -C ₂ haloalkyl-C ₃ -C ₈ cycloalkyl, C ₃ aminylalkylcycloalkyl, C ₁ -C ₂ alkyl-C ₃ cycloalkyl and C ₁ -C ₂ alkyl-4-membered heterocyclylalkyl containing a nitrogen or oxygen heteroatom; R ⁵ means H; and R ⁶ in each case independently represents halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, cyano, C ₁ -C ₆ hydroxyalkyl or C ₁ -C ₆ haloalkyl. 2. The compound according to claim 1, wherein R ² is 2-methylpropyl, optionally substituted with hydroxyl. 3. A compound according to any one of paragraphs 1, 2, where R ² has one of the following structures: 4. A compound according to any one of paragraphs 1-3, wherein R ⁴ has one of the following structures: 5. The compound according to claim 1, wherein A is phenyl. 6. The compound according to claim 1, wherein A is saturated cyclohexyl. 7. The compound according to claim 1, wherein A is pyridinyl. 8. The compound according to claim 1, wherein A is pyrimidinyl. 9. A compound according to any one of claims 1 to 8, wherein A is unsubstituted. 10. The compound according to claim 1, wherein R ⁶ is halogen. 11. The compound according to claim 10, wherein the halogen is chlorine or fluorine. 12. The compound according to claim 1, wherein R ⁶ is C ₁ -C ₆ hydroxyalkyl. 13. The compound according to claim 12, wherein the C ₁ -C ₆ hydroxyalkyl is -CH ₂ CH ₂ OH. 14. The compound according to claim 1, wherein R ⁶ is a cyano group. 15. The compound according to claim 1, wherein R ⁶ is a C ₁ -C ₆ alkoxy group. 16. The compound according to claim 15, wherein the C ₁ -C ₆ alkoxy group is a methoxy group. 17. A compound according to any one of paragraphs 1-16, wherein A has one of the following structures: 18. A compound according to any one of claims 1 to 17, wherein the compound has the following structure (IA): or a pharmaceutically acceptable salt thereof, where: R ^2a means or _C3 - _C4 cycloalkyl optionally substituted with one or more substituents selected from the group consisting of hydroxyl and _C1 - _C6 alkyl; R ^4a is isoxazolyl optionally substituted with one or more substituents selected from the group consisting of C ₁ -C ₆ haloalkyl, C ₃ -C ₈ cycloalkyl or C ₃ -C ₈ haloalkylcycloalkyl. 19. The compound according to claim 18, wherein R ^2a has one of the following structures: 20. The compound according to claim 18 or 19, wherein R ^4a has one of the following structures: 21. The compound of claim 1, wherein the compound has one of the following structures: or a pharmaceutically acceptable salt thereof. 22. The compound of any one of claims 1-21, wherein the compound is a modulator of the NLRP3 inflammasome. 23. The compound of any one of claims 1-21, wherein the compound is an NEK7 inhibitor. 24. A pharmaceutical composition for inhibiting NEK7, modulating NLRP3 activity, or both, comprising a therapeutically effective amount of a compound according to any one of claims 1-21 and a pharmaceutically acceptable carrier, diluent, or excipient. 25. A method for treating an NLRP3-mediated disorder comprising administering a therapeutically effective amount of a compound according to any one of claims 1-21 or a pharmaceutical composition according to claim 24 to a subject in need thereof. 26. The method of claim 25, wherein the disorder is selected from the group consisting of the following: autoimmune, inflammatory disorders, cardiovascular diseases, neurodegenerative disorders, allergies, asthma, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, transplantation, sperm motility, transplant rejection, lung lesions, respiratory diseases, and ischemic conditions. 27. The method of claim 25 or 26, wherein the disorder is selected from the group consisting of type II diabetes, atherosclerosis, Alzheimer's disease, fatty liver disease, asthma, psoriasis, obesity, acute and chronic tissue injury caused by infection, gout, arthritis, macular degeneration, enteritis, hepatitis, peritonitis, silicosis, UV-induced sunburn, contact hypersensitivity, sepsis, cancer, neurodegenerative disease, multiple sclerosis, Maki-Wells syndrome, and myelodysplastic syndrome (MDS).