Pyrimidin-2-yl-pyrazole compounds as IRAK inhibitors

Abstract

The disclosed embodiments relate to novel interleukin receptor associated kinase (IRAK) inhibitor compounds and compositions comprising such compounds. The compounds may have a structure according to Formula I (I) # imgabs0 #. Methods of making and using the compounds and compositions are also disclosed. The disclosed compounds and / or compositions are useful for the treatment or prevention of IRAK-related diseases or disorders.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of the earlier filing date of U.S. Provisional Patent Application No. 63 / 322,955, filed on March 23, 2022, which is incorporated herein by reference in its entirety. Technical Field

[0003] The present disclosure relates to pyrazole compounds, and embodiments of methods for making and using the compounds, such as for inhibiting interleukin receptor-associated kinases (IRAKs) and for treating diseases and disorders associated with IRAKs. Background Art

[0004] Interleukin-1 receptor associated kinase (IRAK) is an important mediator of signal transduction processes such as toll-like receptor (TLR) and interleukin-1 receptor (IL-1R) signal transduction processes. IRAK has been implicated in regulating signal transduction networks that control inflammation, apoptosis, and cell differentiation. Four IRAK genes (IRAK1, IRAK2, IRAK3, and IRAK4) have been identified in the human genome, and studies have revealed different non-redundant biological roles. IRAK1 and IRAK4 have been shown to exhibit kinase activity. Summary of the invention

[0005] Disclosed herein are embodiments of compounds having a structure according to Formula I, or a pharmaceutically acceptable salt or solvate thereof:

[0006]

[0007] About Formula I, R 1 is H, aliphatic, acyl, heterocyclic, carboxy ester, amide, alkyl phosphoramidate or alkyl phosphate, such as H, alkyl or -alkyl OP(O)(OR) 2 , for example -CH(CH 3 )OP(O)(OR) 2 or -CH 2 OP(O)(OR) 2 , wherein each OR is -OH, -Oalkyl, -Oaryl, -Oheteroaryl, -Oaralkyl or -O - M + , where M + is a counter ion with a single positive charge, and R 2 C 1-6 In some embodiments, R 2 C 1-4 In some embodiments, the compound has a structure according to Formula II or a pharmaceutically acceptable salt or solvate thereof.

[0008] In one embodiment, R 1 is H. In another embodiment, R 1 C 1-6 In another embodiment, R 1 -CH 2 OP(O)(OR) 2 , and can be -CH 2 OP(O)(OH) 2 、-CH 2 OP(O)(OC 1-6 alkyl) 2 or -CH 2 OP(O)(O - M + ) 2 , such as -CH 2 OP(O)(O - Na + ) 2 In another embodiment, R 1 -CH(CH 3 )OP(O)(OR) 2 , and may be -CH(CH 3 )OP(O)(OH) 2 、-CH(CH 3 )OP(O)(OC 1-6 alkyl) 2 or -CH(CH 3 )OP(O)(O - M + ) 2 , such as -CH(CH 3 )OP(O)(O - Na + ) 2 .

[0009] The compound may be in free base form or in salt form, such as a co-crystal, for example a tartrate co-crystal or a triple salt co-crystal.

[0010] Also disclosed herein is a pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable excipient.

[0011] Disclosed herein is a method for inhibiting an IRAK enzyme. The method may include contacting the enzyme with an effective amount of a compound disclosed herein. In some embodiments, contacting the enzyme includes administering the compound to a subject.

[0012] Also disclosed herein is a method for treating a disease or illness requiring an IRAK inhibitor in a subject. The method may include administering an effective amount of a compound disclosed herein or a pharmaceutical composition thereof to the subject. The disease or illness may include autoimmune diseases, inflammatory conditions, cardiovascular diseases, neurodegenerative conditions, allergic conditions, multiple organ failure, kidney disease, platelet aggregation, hyperproliferative conditions, transplantation, sperm motility, erythrocyte deficiency, transplant rejection, lung injury, respiratory disease, ischemic disease, bacterial infection, viral infection, immunomodulatory conditions, or a combination thereof. In some embodiments, the disease or illness includes aplastic anemia, atopic dermatitis, pustular psoriasis, palmoplantar pustulosis, primary biliary cirrhosis, pyoderma, sclerosing cholangitis, systemic juvenile idiopathic arthritis, suppurative hidradenitis, cytokine release syndrome, or myelodysplastic syndrome (MDS).

[0013] In some embodiments, the disease or illness includes lymphoid tumors. The lymphoid tumor can be selected from myeloproliferative neoplasms (MPNs) excluding polycythemia vera, myeloid / lymphoid tumors with PDGFRA rearrangement, myeloid / lymphoid tumors with PDGFRB rearrangement, myeloid / lymphoid tumors with FGFR1 rearrangement, myeloid / lymphoid tumors with PCM1-JAK2, myelodysplasia / myeloproliferative neoplasms (MDS / MPNs), myeloid sarcomas, myeloproliferation associated with Down syndrome, blastic plasmacytoid dendritic cell tumors, B lymphoblastic leukemia / lymphoma; and / or T lymphoblastic leukemia / lymphoma. In some embodiments, the lymphoid tumor is a myeloproliferative tumor selected from the following: chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), primary myelofibrosis (PMF), essential thrombocythemia, chronic eosinophilic leukemia or a combination thereof. In certain embodiments, the lymphoid tumor is chronic myeloid leukemia.

[0014] In any embodiment, the method may further comprise identifying a subject having the lymphoid tumor. In certain embodiments, the lymphoid tumor is chronic myelomonocytic leukemia, and identifying the subject comprises identifying a persistent peripheral blood monocytosis of ≥ 1×10 9 / L and monocytes accounting for ≥10% of the white blood cell (WBC) differential count and no observed rearrangements in the PDGFRA, PDGFRB, or FGFR1 genes and the PCM1-JAK2 fusion gene.

[0015] The foregoing and other objects, features and advantages of the present technology will become apparent from the following detailed description. DETAILED DESCRIPTION

[0016] I. Definitions

[0017] The following explanations of terms and methods are provided to better describe the present disclosure and guide those of ordinary skill in the art to practice the present disclosure. The singular forms "a / an" and "the" refer to one / a or more than one / a, unless the context clearly indicates otherwise. The term "or" refers to a single element of the stated alternative elements or a combination of two or more elements, unless the context clearly indicates otherwise. As used herein, "comprising" means "including". Therefore, "comprising A or B," means "comprising A, B, or A and B" without excluding other elements. All references cited herein (including patents and patent applications) are incorporated by reference.

[0018] Unless otherwise indicated, all numbers expressing quantities of components, molecular weights, percentages, temperatures, times, etc., as used in the specification or claims will be understood to be modified by the term "about". Therefore, unless otherwise indicated, either explicitly or implicitly, the numerical parameters set forth are approximate values ​​that may depend on the desired desired properties and / or the limits of detection under standard test conditions / methods. When directly or explicitly distinguishing an embodiment from the prior art being discussed, the quantities of the embodiments are not approximate unless the word "about" is cited.

[0019] Unless otherwise explained, all technical and scientific terms used herein have the same meaning as those of ordinary skill in the art to which the present disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the following describes suitable methods and materials. The materials, methods and examples are merely illustrative and are not intended to be limiting.

[0020] When depicting or describing chemical structures, unless otherwise clearly stated, it is assumed that all carbons include hydrogen so that each carbon meets the valence of four. For example, in the structure on the left hand side of the following schematic diagram, it is implied that there are nine hydrogen atoms. The nine hydrogen atoms are depicted in the right hand structure.

[0021]

[0022] Sometimes specific atoms in a structure are described in the formula herein as having hydrogen or a hydrogen atom, such as -CH 2 CH 2 -. Those of ordinary skill in the art will appreciate that the techniques described above are common in the chemical arts to provide brevity and simplicity in the description of organic structures.

[0023] One of ordinary skill in the art will appreciate that the definitions can be combined to further describe a particular compound. For example, hydroxyaliphatic refers to an aliphatic group substituted with a hydroxy (-OH) group, and haloalkylaryl refers to an aryl group substituted with an alkyl group, wherein the alkyl group is also substituted with a halogen, and wherein the point of attachment to the parent structure is through the aryl portion, since aryl is the base name for the substituent.

[0024] As used herein, the term "substituted" refers to all subsequent modifications in the term, for example, in the term "substituted aryl C 1-8 Alkyl substitution can occur on the aromatic C 1-8 The "C" of the alkyl 1-8 The term "alkyl" refers to an "alkyl" group, an "aryl" group, or both. By way of further example, alkyl includes substituted cycloalkyl.

[0025] When used to modify a specified group or part, "substituted" means that at least one and perhaps two or more hydrogen atoms of the specified group or part are independently replaced by the same or different substituent groups defined below. In a specific embodiment, a group, part or substituent may be substituted or unsubstituted unless explicitly defined as "unsubstituted" or "substituted". Therefore, any group specified herein may be unsubstituted or substituted. In a specific embodiment, a substituent may or may not be explicitly defined as substituted, but is still considered to be optionally substituted. For example, an "alkyl" substituent may be unsubstituted or substituted, but an "unsubstituted alkyl" may not be substituted.

[0026] In one embodiment, a substituted group has 1 substituent, 2 substituents, 2 substituents, 2 substituents, or 4 substituents.

[0027] In addition, in the embodiment that the substituted substituent of group or part replaces, the nesting of the substituent of this type of replacement is limited to three, thus prevents the formation of polymer.Therefore, in the group or part comprising the first group, the first group is the substituent on the second group, the second group itself is the substituent on the third group, the group or part is connected to the parent structure, and the first (outermost) group can only be replaced by unsubstituted substituent.For example, in the group comprising-(heteroaryl-1)-(heteroaryl-2)-(heteroaryl-3), heteroaryl-3 can only be replaced by its own unsubstituted substituent.

[0028] "Aliphatic" refers to a substantially hydrocarbon-based group or moiety, including alkyl, alkenyl, alkynyl, cyclic versions thereof, such as cycloalkyl, cycloalkenyl or cycloalkynyl, and further including straight and branched arrangements, and all stereo and positional isomers. Unless otherwise expressly stated, aliphatic groups contain one to twenty-five carbon atoms; e.g., one to fifteen, one to ten, one to six, or one to four carbon atoms, and cycloaliphatics contain three to twenty-five carbon atoms; e.g., three to fifteen, three to ten, three to six, or three to four carbon atoms.

[0029] "Acyl" refers to the group -C(O)R, where R is H, aliphatic, heteroaliphatic, heterocyclic, or aryl. Exemplary acyl moieties include, but are not limited to, -C(O)H, -C(O)alkyl, -C(O)C 1 -C 6 Alkyl, -C(O)C 1 -C 6 Haloalkyl-C(O)cycloalkyl, -C(O)alkenyl, -C(O)cycloalkenyl, -C(O)aryl, -C(O)heteroaryl or -C(O)heterocyclyl. Specific examples include -C(O)H, -C(O)Me, -C(O)Et or -C(O)cyclopropyl.

[0030] "Alkyl" refers to a saturated aliphatic hydrocarbon group having 1 to 25 carbon atoms, typically 1 to 10 carbon atoms, such as 1 to 6 carbon atoms (C 1 -C 6 The alkyl moiety may be substituted or unsubstituted. For example, this term includes straight and branched chain hydrocarbon groups, cycloalkyl groups, and combinations thereof, such as -CH 2 Cyclopropyl, unless otherwise indicated. Cycloalkyl refers to a cyclic aliphatic group having 3 to 15 carbon atoms, typically 3 to 8 carbon atoms, 3 to 6 carbon atoms, or 3 to 4 carbon atoms. The cycloalkyl group may be a monocyclic ring (e.g., cyclohexyl), or may contain multiple rings, such as fused, bridged, or spirocyclic systems, at least one of which is aliphatic, provided that the point of attachment is through an aliphatic region of the cycloalkyl group. Exemplary alkyl groups include, but are not limited to, methyl (CH 3 ), ethyl (-CH 2 CH 3 ), n-propyl (-CH 2 CH 2 CH 3 ), isopropyl (-CH(CH 3 ) 2 ), n-butyl (-CH 2 CH 2 CH 2 CH 3 ), isobutyl (-CH 2 CH 2(CH 3 ) 2 ), sec-butyl (-CH(CH 3 )(CH 2 CH 3 ), tert-butyl (-C(CH 3 ) 3 ), n-pentyl (-CH 2 CH 2 CH 2 CH 2 CH 3 ), neopentyl (-CH 2 C(CH 3 ) 3 ), cyclopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, -CH 2 Cyclopropyl, -CH 2 Cyclobutyl, -CH 2 Cyclopentyl or -CH 2 Cyclohexyl.

[0031] "Amide" refers to the group -N(H)acyl or -C(O)amino.

[0032] "Araliphatic" refers to an aromatic group attached to the parent through an aliphatic moiety. Araliphatics include aralkyl or arylalkyl groups such as benzyl and phenylethyl.

[0033] "Aralkyl" refers to an aryl group attached to the parent through an alkyl moiety. Aralkyl includes groups such as benzyl and phenylethyl.

[0034] Unless otherwise indicated, "aryl" refers to an aromatic group having 5 to 15 ring atoms, the aromatic group having a single ring (e.g., phenyl) or multiple fused rings in which at least one ring is aromatic (e.g., naphthyl). For groups having multiple rings, at least one of which is aromatic and one is not, such groups are referred to as "aryl" with the proviso that the point of attachment to the remainder of the compound is through an atom of the aromatic portion of the aryl. Aryl groups may be monocyclic, bicyclic, tricyclic, or tetracyclic. Unless otherwise indicated, aryl groups may be substituted or unsubstituted.

[0035] "Carboxyl" or "carboxy" or "carboxylate" refers to a -CO 2 H、-C(O)O - or a salt thereof.

[0036] "Carboxyl ester" or "carboxy ester" refers to the group -C(O)OR, where R is aliphatic, heteroaliphatic and heterocyclic, including aryl and heteroaryl.

[0037] "Cycloaliphatic" refers to a cyclic aliphatic group having a single ring (e.g., cyclohexyl) or multiple rings (such as a fused, bridged, or spirocyclic system, at least one of which is aliphatic, provided that the point of attachment is through an aliphatic region of the cycloaliphatic group). Cycloaliphatics include saturated and unsaturated systems, including cycloalkyl, cycloalkenyl, and cycloalkynyl. Exemplary cycloaliphatic groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, or cyclohexenyl.

[0038] "Halo," "halide," or "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0039] "Haloalkyl" refers to an alkyl moiety substituted with one or more halogens. Exemplary haloalkyl moieties are CF 3 .

[0040] "Heterocyclyl" and "heterocycle" refer to aromatic and non-aromatic ring systems, and more specifically to stable three to fifteen-membered ring moieties containing carbon atoms and at least one (such as one to five) heteroatoms. The heterocyclyl moiety may be a monocyclic moiety, or may contain multiple rings, such as in a bicyclic or tricyclic ring system, provided that at least one of the rings contains a heteroatom. Such polycyclic moieties may include fused or bridged ring systems and spirocyclic systems; and the nitrogen, phosphorus, carbon, silicon or sulfur atoms in the heterocyclyl moiety may be optionally oxidized to various oxidation states. For convenience, nitrogen, particularly but not exclusively, those defined as cyclic aromatic nitrogens are not meant to include their corresponding N-oxide forms, although not explicitly defined as such in specific examples. In addition, cyclic nitrogen atoms may be optionally quaternized. Heterocycles include heteroaryl or aromatic heterocyclyl moieties and non-aromatic heterocyclyl moieties (which are partially or fully saturated heterocyclyl rings, such as heterocycloalkyl).

[0041] Unless otherwise indicated, "heteroaryl" refers to an aromatic group or moiety having 5 to 15 ring atoms that contains at least one carbon atom and at least one heteroatom (such as N, S, O, P or Si). The heteroaryl or moiety can contain a single ring (e.g., pyridyl, pyrimidinyl or pyrazolyl) or multiple fused rings (e.g., indolyl, benzopyrazolyl or pyrazolopyridinyl). For example, the heteroaryl or moiety can be monocyclic, bicyclic, tricyclic or tetracyclic. Unless otherwise indicated, the heteroaryl or moiety can be substituted or unsubstituted.

[0042] "Heterocycloalkyl" refers to a stable three to fifteen-membered non-aromatic ring portion containing at least one carbon atom and usually multiple carbon atoms and at least one (such as one to five) heteroatom. The one or more heteroatoms may be one or more nitrogen, phosphorus, oxygen, silicon or sulfur atoms. The heterocycloalkyl portion may be a monocyclic portion, or may contain multiple rings, such as in a bicyclic or tricyclic ring system, provided that at least one of the rings contains a heteroatom. Such polycyclic portions may include fused or bridged ring systems and spirocyclic systems; and any nitrogen, phosphorus, carbon, silicon or sulfur atom in the heterocycloalkyl portion may be optionally oxidized to various oxidation states, unless explicitly excluded or excluded by context. For convenience, nitrogen, particularly but not exclusively, those defined as cyclic aromatic nitrogens are not meant to include their corresponding N-oxide forms, although not explicitly defined as such in specific examples. In addition, cyclic nitrogen atoms may be optionally quaternized.

[0043] Examples of heterocycloalkyl include, but are not limited to, tetrahydroisoquinolinyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, pyrrolidinyl, 4-piperidonyl, dihydropyridinyl, tetrahydropyridinyl, morpholinyl, diazabicycloheptane, diazapane, diazepine, tetrahydrofuranyl, and tetrahydropyranyl.

[0044] "Phosphate" refers to the group -OP(O)(OR') 2 , wherein each -OR' is independently -OH, -O-aliphatic (such as -O-alkyl), -O-aryl or -O-aralkyl, or -OR' is -O - M + , where M + M is a counter ion having a single positive charge as disclosed herein. For example, each M + Can be alkaline ions such as K + 、Na + , Li + ; ammonium ions, such as + N(R") 4 , wherein each R" is independently H, an aliphatic such as an alkyl, hydroxyalkyl or a combination thereof, a heterocyclic group or an aryl group; an amino acid such as arginine or lysine; an amino sugar such as meglumine; or an alkaline earth metal ion such as [Ca 2+ ] 0.5 , [Mg 2+ ] 0.5 or[Ba 2+ ] 0.5 Alkyl phosphate refers to the group -alkyl-phosphate, such as -CH 2 OP(O)(OR') 2 or -CH 2 (CH3 )OP(O)(OR') 2 , for example, -CH 2 OP(O)(O-isopropyl) 2 、-CH 2 OP(O)(OH)(O-tert-butyl), -CH 2 OP(O)(O-tert-butyl) 2 、-CH 2 OP(O)(OCH 2 OCO 2 Isopropyl) 2 、-CH 2 OP(O)(OH) 2 or a salt thereof, such as -CH 2 OP(O)(O - Na + ) 2 、-CH 2 OP(O)(O - ) 2 Mg 2+ or -CH 2 OP(O)(OH)(O - Na + )

[0045] "Phosphoramidate" refers to the group -OP(O)(OR')(N(R') 2 ), wherein each R' is independently H, an aliphatic (such as an alkyl, aryl or aralkyl) group, or -OR' is -O - M + , and where M + M is a counter ion having a single positive charge as disclosed herein. For example, each M + Can be alkaline ions such as K + 、Na + , Li + ; ammonium ions, such as + N(R") 4 , wherein each R" is independently H, an aliphatic such as an alkyl, hydroxyalkyl or a combination thereof, a heterocyclic group or an aryl group; an amino acid such as arginine or lysine; an amino sugar such as meglumine; or an alkaline earth metal ion such as [Ca 2+ ] 0.5 , [Mg 2+ ] 0.5 or[Ba 2+ ] 0.5 Alkyl phosphoramidate refers to the group -alkyl-phosphoramidate, such as, for example, -CH 2 OP(O)(OR')(N(R' 2 )) or -CH 2(CH 3 )OP(O)(OR')(N(R' 2 )), such as -CH 2 OP(O)(O-phenyl)[NHC(CH 3 )CO 2 isopropyl] or -CH 2 OP(O)(OH)(N(H)alkyl) or a salt thereof, such as -CH 2 OP(O)(O - Na + )(N(H)alkyl).

[0046] "Patient" or "subject" refers to a mammal or other animal, particularly a human. Thus, the disclosed methods are applicable to both human therapy and veterinary applications.

[0047] "Pharmaceutically acceptable excipient" refers to a substance other than an active ingredient, which is included in the formulation of an active ingredient. As used herein, an excipient can be incorporated into the particles of a pharmaceutical composition, or it can be physically mixed with the particles of a pharmaceutical composition. For example, an excipient can be used to dilute an active agent and / or change the properties of a pharmaceutical composition. An excipient can include, but is not limited to, an antiadhesive, a binder, a coating, an enteric coating, a disintegrant, a flavoring agent, a sweetener, a colorant, a lubricant, a glidant, an adsorbent, a preservative, an adjuvant, a carrier or a vehicle. An excipient can be starch and modified starch, cellulose and cellulose derivatives, sugars and their derivatives (such as disaccharides, polysaccharides and sugar alcohols), proteins, synthetic polymers, cross-linked polymers, antioxidants, amino acids or preservatives. Exemplary excipients include, but are not limited to, magnesium stearate, stearic acid, vegetable fats, sucrose, lactose, starch, hydroxypropylcellulose, hydroxypropylmethylcellulose, xylitol, sorbitol, maltitol, gelatin, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), tocopheryl polyethylene glycol 1000 succinate (also known as vitamin E TPGS or TPGS), carboxymethylcellulose, dipalmitoylphosphatidylcholine (DPPC), vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium, cysteine, methionine, citric acid, sodium citrate, methylparaben, propylparaben, sugar, silicon dioxide, talc, magnesium carbonate, sodium starch glycolate, tartrate, aspartame, benzalkonium chloride, sesame oil, propyl gallate, sodium metabisulfite, or lanolin.

[0048] "Adjuvant" is an excipient that changes the effect of other agents (usually active ingredients). Adjuvants are typically pharmaceutical and / or immunological agents. Adjuvants can change the effect of active ingredients by increasing the immune response. Adjuvants can also serve as stabilizers for formulations. Exemplary adjuvants include, but are not limited to, aluminum hydroxide, alum, aluminum phosphate, killed bacteria, squalene, detergents, cytokines, paraffin oil, and combination adjuvants (such as Freund's complete adjuvant or Freund's incomplete adjuvant).

[0049] "Pharmaceutically acceptable carrier" refers to an excipient that acts as a carrier or vehicle, such as a suspending aid, a solubilizing aid, or a nebulizing aid. Remington: The Science and Practice of Pharmacy, The University of the Sciences in Philadelphia, Editor, Lippincott, Williams, & Wilkins, Philadelphia, PA, 21st edition (2005), which is incorporated herein by reference, describes exemplary compositions and formulations suitable for drug delivery of one or more therapeutic compositions and additional pharmaceutical agents.

[0050] In general, the properties of the carrier will depend on the specific mode of administration adopted. For example, parenteral preparations generally include injectable fluids, which include pharmaceutically or physiologically acceptable fluids, such as water, saline, balanced salt solutions, aqueous glucose, glycerol or the like as vehicles. In some instances, pharmaceutically acceptable carriers may be sterile to be suitable for administration to a subject (e.g., by parenteral, intramuscular or subcutaneous injection). In addition to biologically neutral carriers, the pharmaceutical composition to be administered may contain a small amount of non-toxic auxiliary substances, such as wetting agents or emulsifiers, preservatives and pH buffers, such as sodium acetate or sorbitan monolaurate.

[0051] "Pharmaceutically acceptable salts" refers to pharmaceutically acceptable salts of compounds derived from various organic and inorganic counterions as would be known to one of ordinary skill in the art, and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when the molecule contains a basic functional group, salts of organic or inorganic acids such as hydrochlorides, hydrobromides, tartrates, methanesulfonates, acetates, maleates, oxalates, and the like. "Pharmaceutically acceptable acid addition salts" are a subset of "pharmaceutically acceptable salts" that retain the biological effectiveness of the free bases when formed by an acid partner. In particular, the disclosed compounds form salts with various pharmaceutically acceptable acids, including but not limited to inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., and organic acids such as formic acid, acetic acid, adipic acid, aspartic acid, trifluoroacetic acid, propionic acid, gentisic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, benzenesulfonic acid, hydroxyethanesulfonic acid, lactic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, xinafoic acid, etc. "Pharmaceutically acceptable base addition salts" are a subset of "pharmaceutically acceptable salts" derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, etc. Exemplary salts are ammonium, potassium, sodium, calcium and magnesium salts. Salts derived from pharmaceutically acceptable organic bases include, but are not limited to, primary, secondary and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines and basic ion exchange resins (such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, tris(hydroxymethyl)aminomethane (Tris), ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc.). Exemplary organic bases are isopropylamine, diethylamine, tris(hydroxymethyl)aminomethane (Tris), ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. (See, eg, SM Berge et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977; 66: 1-19, which is incorporated herein by reference.) In certain disclosed embodiments, the pyrazole compound can be a formate salt or a sodium salt.

[0052] An "effective amount" with respect to a compound or composition refers to an amount of the compound or composition sufficient to achieve a specific desired result, such as inhibiting a protein or enzyme, particularly an interleukin-1 receptor-associated kinase; inducing a desired biological or medical response in a tissue, system, subject, or patient; treating or preventing a specific condition or disease; ameliorating or eradicating one or more of its symptoms; and / or preventing the occurrence of a disease or condition. The amount of a compound that constitutes an "effective amount" may vary depending on the compound, the desired result, the disease state and its severity, the age of the patient to be treated, and the like.

[0053] "Solvate" refers to a complex formed by the combination of solvent molecules with molecules or ions of a solute. The solvent may be an organic solvent, an inorganic solvent, or a mixture of the two. Exemplary solvents include, but are not limited to, alcohols such as methanol, ethanol, propanol; amides such as N,N-dialiphatic amides such as N,N-dimethylformamide; tetrahydrofuran; alkyl sulfoxides such as dimethyl sulfoxide; water; and combinations thereof. When combined with a pharmaceutically acceptable or unacceptable solvent (such as water, ethanol, etc.), the compounds described herein can exist in unsolvated forms as well as solvated forms. The solvated forms of the compounds disclosed herein are within the scope of the embodiments disclosed herein.

[0054] As used herein, "treating" or "treatment" refers to treating a target disease or disorder in a patient or subject, particularly a human suffering from the target disease or disorder, and includes, by way of example and without limitation:

[0055] (i) inhibiting the disease or disorder, for example arresting or slowing its progression;

[0056] (ii) ameliorating the disease or disorder, e.g., causing regression of the disease or disorder or a symptom thereof; or

[0057] (iii) stabilizing the disease or disorder.

[0058] As used herein, "preventing" or "preventing" relates to preventing a disease or disorder from occurring in a patient or subject, particularly when such patient or subject is susceptible to the disorder but has not yet been diagnosed as having the disorder.

[0059] As used herein, the terms "disease," "disorder," and "condition" may be used interchangeably or may differ in that a particular malady or condition may have no known causative factor (so a cause has not yet been determined) and is therefore not yet considered a disease but merely an undesirable condition or syndrome in which a more or less specific constellation of symptoms is identified by a clinician.

[0060] The above definitions and the following formulae are not intended to include impermissible substitution patterns (eg, a methyl group substituted with 5 fluoro groups). One of ordinary skill in the art would readily recognize such impermissible substitution patterns.

[0061] Any group mentioned herein may be optionally substituted with at least one (possibly two or more) substituents as defined herein. That is, unless the context otherwise indicates or a particular structural formula excludes substitution, the substituted group has at least one (possibly two or more) replaceable hydrogen, which is replaced by one or more substituents as defined herein.

[0062] Those of ordinary skill in the art will appreciate that compounds can exhibit tautomerism, conformational isomerism, geometric isomerism, and / or optical isomerism. For example, certain disclosed compounds may include one or more chiral centers and / or double bonds, and therefore may exist as stereoisomers, such as double bond isomers (i.e., geometric isomers), enantiomers, diastereomers, and mixtures thereof (such as racemic mixtures). As another example, certain disclosed compounds may exist in several tautomeric forms, including enol forms, keto forms, and mixtures thereof. Since the various compound names, formulas, and compound diagrams in the specification and claims may represent only one of possible tautomerism, conformational isomerism, optical isomerism, or geometric isomerism, it will be understood that the disclosed compounds encompass any tautomerism, conformational isomerism, optical isomerism, and / or geometric isomerism of the compounds described herein, as well as mixtures of these various different isomeric forms. In cases of limited rotation (eg, around an amide bond or between two directly connected rings such as pyrazole and pyridyl rings), atropisomers are also possible and are also specifically included in the compounds of the present disclosure.

[0063] Specific examples of compounds disclosed herein include one or more asymmetric centers; therefore, these compounds can exist in different stereoisomeric forms. Therefore, compounds and compositions can be provided as individual pure enantiomers or diastereomers, or as stereoisomer mixtures (including racemic mixtures). In certain embodiments, the compounds disclosed herein are synthesized or purified in a substantially enantiomerically pure form, such as at least 90% enantiomeric excess, 95% enantiomeric excess, 97% enantiomeric excess, 98% enantiomeric excess, 99% enantiomeric excess, 99.5% enantiomeric excess, or greater than 99.5% enantiomeric excess, such as in enantiomerically pure form.

[0064] In any embodiment, any or all hydrogens present in the compound or in a particular group or moiety within the compound may be replaced by deuterium or tritium. Thus, the recitation of alkyl includes deuterated alkyls in which one to the maximum number of hydrogens present may be replaced by deuterium. For example, ethyl may be C 2 H 5 or C 2 H 5 , in which 1 to 5 hydrogens are replaced by deuterium.

[0065] II.Compounds

[0066] Disclosed herein are compounds, methods for preparing the compounds, and methods for using the compounds. In one embodiment, the disclosed compounds are kinase inhibitors, particularly tyrosine kinase inhibitors. In a specific embodiment, the compounds can be used to block one or more cytokine signaling pathways (such as IL-17 signaling pathways). For certain embodiments, the disclosed compounds can be used to treat disorders for which inhibition of the interleukin-1 receptor associated kinase (IRAK) pathway is useful for treatment. In some embodiments, the compounds directly inhibit IRAK proteins, such as IRAK1, IRAK2, IRAK3, and / or IRAK4.

[0067] Exemplary compounds within the scope of the present disclosure have the general formula I

[0068]

[0069] or a pharmaceutically acceptable salt or solvate thereof.

[0070] About Formula I, R 1 is H, aliphatic, acyl, heterocyclic, carboxy ester, amide, alkyl phosphoramidate or alkyl phosphate. 1 Not H, or alternatively, R 1 is hydrogen. 1 When R is hydrogen, the compound may be a free base or in salt form. 1 is an alkyl, acyl, carboxylate, amide, non-aromatic heterocyclic group, alkyl phosphoramidate or alkyl phosphate. A person of ordinary skill in the art understands that, for example, when administered to a subject, wherein R 1 Compounds other than H can act as 1 A prodrug of a compound of H.

[0071] R 2 C 1-6 Alkyl groups, such as C 1-4 alkyl.

[0072] In some embodiments, R 2 It is ethyl.

[0073] In some embodiments, R 1 H, C 1-4 Alkyl phosphate, C 1-4 Alkyl phosphoramidate, C 1-6 Alkyl, C 1-6 Acyl, -C(O)OC 1-6 Aliphatic, -C(O)N(R b ) 2 or a 5-membered or 6-membered non-aromatic heterocyclic group, but in certain embodiments, R 1 Not H, or R 1 is H and the compound is a salt.

[0074] About R 1 Part C 1-6 The alkyl portion may be unsubstituted or it may be substituted, for example, by a 5- or 6-membered non-aromatic heterocyclic group, OH, -OC(O)-R a 、-N(R b ) 2 、-OC(O)-R c , carboxyl or a combination thereof;

[0075] The C 1-6 The acyl moiety may be unsubstituted or it may be replaced by -C(O)OC 1-4 Alkyl, -C(O)OC 1-4 Alkyl-N(R b ) 2 、N(R b ) 2 、-NHC(O)C 1-4 Alkyl or combination thereof substitution;

[0076] The 5-membered or 6-membered heterocyclic moiety may be a 5-membered or 6-membered oxygen-containing heterocyclic group, and / or may be substituted by a hydroxyl group, a hydroxymethyl group or a combination thereof; or

[0077] The -C(O)OC 1-6 The aliphatic group may be optionally substituted with -OC(O)C 1-4 Alkyl or N(R b ) 2 Substituted -C(O)OC 1-6 Alkyl, or the -C(O)OC 1-6 The aliphatic group may be optionally replaced by C 1-4 Alkyl-substituted-C(O)OC 3-6 Cycloalkyl.

[0078] In any embodiment, each R a is independently a 5-membered non-aromatic heterocyclic group,2 N(R b ) 2 Substituted aryl, C substituted by carboxyl 3-6 Cycloalkyl, C 1-6 Alkoxy, unsubstituted C 1-6 Alkyl, or N(R b ) 2 , carboxyl, carboxyl ester, -OC 1-6 Acyl, -NHC(O)(NH 2 )C 1-6 Alkyl or -(OCH 2 CH 2 ) 1-8 N(R b ) 2 One or more (such as 1, 2 or 3) of C 1-6 alkyl;

[0079] Each R b are independently H, unsubstituted C 1-6 Alkyl, -N(R g ) 2 Substituted C 1-6 Alkyl, carboxyl ester or 5-membered or 6-membered non-aromatic heterocyclic group, or two R b Together with the nitrogen to which they are attached, they form a g ) Intermittent C 3-6 A non-aromatic heterocyclic moiety, wherein R g H or C 1-4 Alkyl; and

[0080] -OC(O)-R c Derived from amino acids, where -OC(O)-R c The -OC(O)- portion corresponds to the acid portion of the amino acid, and R c Contains -N(R b ) 2Or a nitrogen-containing non-aromatic heterocyclic group, such as a 5-membered or 6-membered unsaturated nitrogen-containing heterocyclic group, for example a pyrrolidinyl group. The amino acid can be any amino acid, such as a naturally occurring amino acid, and can be selected from the following amino acids: glycine, valine, alanine, leucine, isoleucine, methionine, phenylalanine, tryptophan, tyrosine, serine, threonine, asparagine, glutamine, arginine, histidine, lysine, aspartic acid, glutamic acid, cysteine ​​or proline. Those of ordinary skill in the art will understand that in the case where an amino acid contains one or more chiral centers, all enantiomers, diastereomers and / or mixtures thereof are considered. For example, an amino acid can be an L-amino acid, a D-amino acid or a mixture thereof. In some embodiments, an amino acid is an L-amino acid. And in certain embodiments, -OC(O)-R c -OC(O)CH(NH 2 )R d , or -OC(O)-(CH 2 ) 1-2 C(NH 2 )CO 2 H, where R d is an amino acid side chain, and / or may be H, -CH 3 , isopropyl, -CH 2 CH(CH 3 ) 2 、-CH(CH 3 )Et, -CH 2 CH 2 SCH 3 , -CH 2 OH, -CH(OH)CH 3 、-CH 2 C(O)NH 2 、-CH 2 CH 2 C(O)NH 2 、-CH 2 SH, -CH 2 CH 2 CH 2 NHC(O)(NH)NH 2 , -CH 2 CH 2 CH 2 CH 2 NH 2 、-CH 2 CO 2 H or CH 2 CH 2 CO 2 H.

[0081] In some embodiments, R 1 is H, alkyl or -alkylOP(O)(OR) 2 , wherein each OR is independently -OH, -Oalkyl, -Oaryl, -Oheteroaryl, -Oaralkyl or -O - M + , where M + is a counter ion with a single positive charge. Each OR can independently be -OH, -O(C 1-6 Alkyl), -OC 6 Aryl, -O(3-15 membered heteroaryl), -OC 7 Aralkyl or -O - M + And in some embodiments, the two OR moieties are different, but in other embodiments, the two OR moieties are the same.

[0082] In some embodiments, R 1 H, C 1-6 Alkyl or -C 1-4 Alkyl OP(O)(OR) 2 , such as H, C 1-6 Alkyl, -CH 2 OP(O)(OR) 2 or -CH(CH 3 )OP(O)(OR) 2 , and in certain embodiments, R 1 H, C 1-6 Alkyl or -CH 2 OP(O)(OR) 2 .

[0083] In certain embodiments, the compound has a structure according to Formula II or a pharmaceutically acceptable salt or solvate thereof.

[0084]

[0085]

[0086] Regarding Formula II, R 1 As previously defined for Formula I.

[0087] In some embodiments of Formulas I and II, R 1 For H.

[0088] In other embodiments of Formulas I and II, R 1 is an alkyl group, such as C 1-6 Alkyl or C 1-4 The alkyl group may be a methyl group or an ethyl group.

[0089] In some other embodiments of Formulas I and II, R 1 is -alkyl OP(O)(OR) 2 , such as -C 1-4 Alkyl OP(O)(OR) 2 , and can be -CH 2 OP(O)(OR) 2 or -CH(CH 3 )OP(O)(OR) 2 .

[0090] In some embodiments, R 1 -CH 2 OP(O)(OR) 2 In one embodiment, R 1 -CH 2 OP(O)(OH) 2 In another embodiment, R 1 -CH 2 OP(O)(OH)(O - M + In another embodiment, R 1 -CH 2 OP(O)(O - M + ) 2 , and can be -CH 2 OP(O)(O - Na + ) 2 And in another embodiment, R 2 -CH 2 OP(O)(OC 1-6 alkyl) 2 .

[0091] In some embodiments of Formula I and Formula II, R 1 -CH(CH 3 )OP(O)(OR) 2 In one embodiment, R 1 -CH(CH 3 )OP(O)(OH) 2 In another embodiment, R 1 -CH(CH 3 )OP(O)(OH)(O - M + In another embodiment, R 1 -CH(CH 3 )OP(O)(O -M + ) 2 , and may be -CH(CH 3 )OP(O)(O - Na + ) 2 And in another embodiment, R 2 -CH(CH 3 )OP(O)(OC 1-6 alkyl) 2 .

[0092] In one embodiment disclosed herein, the compound comprises an IRAK inhibitory moiety and a means for releasing or delivering the IRAK inhibitory moiety in vivo. 1 In some embodiments where R is not H, 1 Contains means for delivering IRAK inhibitors, such as compound I-1 disclosed herein. Exemplary embodiments of Formula I and Formula II containing such means will be apparent to those skilled in the art upon consideration of the formulas and working examples disclosed herein. For example, embodiments of Formula I and Formula II containing means for delivering an IRAK inhibitory moiety include, but are not limited to, compounds I-2, I-3, I-6, I-7, and I-8.

[0093] In any embodiment, the compound may be in the form of a free base, or it may be a salt. In some embodiments, the salt is an acid addition salt, such as but not limited to an acid addition salt as defined herein. In a specific embodiment, the compound is a tartrate.

[0094] In other embodiments, the compound is in the form of a base addition salt, such as but not limited to a base addition salt as defined herein. In some embodiments, the compound is a sodium salt, such as a disodium salt or a tri-salt.

[0095] Exemplary compounds according to Formula I include, but are not limited to:

[0096]

[0097] Some exemplary compounds according to Formula 1 include:

[0098] I-1: N-(1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol-4-yl)thiazole-4-carboxamide;

[0099] I-2: (4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl dihydrogen phosphate;

[0100] I-3: sodium (4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl phosphate;

[0101] I-4: N-(1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol-4-yl)thiazole-4-carboxamide tartrate;

[0102] I-5: (4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl hydrogen phosphate 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt (trisalt);

[0103] I-6: 1-(4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)ethyl dihydrogen phosphate;

[0104] I-7: sodium 1-(4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)ethyl phosphate; or

[0105] I-8: 1-(4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)ethyl hydrogen phosphate 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt (trisalt).

[0106] In some embodiments, the compound according to any one of Formula I and Formula II or a salt thereof has at least one improved property compared to other IRAK inhibitors. One or more improved properties may include, but are not limited to, physical properties, including, but not limited to, melting point, glass transition temperature, fluidity and / or stability, such as thermal stability, mechanical stability, shelf life, stability against polymorphic transition, etc.; chemical properties, such as, but not limited to, hygroscopic properties, solubility in water and / or organic solvents, reactivity, compatibility with excipients and / or delivery vehicles; and / or pharmacokinetic properties, such as, but not limited to, bioavailability, absorption, distribution, metabolism, excretion, toxicity (including cytotoxicity), dissolution rate, selectivity (such as selectivity for one IRAK protein over a different IRAK protein), activity and / or half-life.

[0107] III. Synthesis

[0108] The disclosed pyrazole compounds can be prepared as exemplified below and will be understood by one of ordinary skill in the art of organic synthesis. An exemplary synthesis may include the following first reaction step according to Scheme 1:

[0109]

[0110] The acetyl compound 2 is reacted with dimethylformamide dimethyl acetal 4 at a temperature suitable to promote the reaction to form an intermediate compound 6. The suitable temperature is generally 85° C. to 130° C. The intermediate compound 6 is then reacted with hydrazine hydrate 8 to form a pyrazole compound 10. The reaction is carried out in a suitable solvent, for example an alcohol such as ethanol, methanol or isopropanol, and is generally heated to, for example, reflux.

[0111] The second reaction step in an exemplary synthesis is provided below according to Scheme 2:

[0112]

[0113] Compound 10 is nitrated to form compound 14 using a suitable nitrating agent or a mixture of agents 12. Suitable nitrating conditions include reacting compound 10 with nitric acid (such as fuming nitric acid) (optionally in the presence of sulfuric acid). Typically, compound 10 and nitric acid are added slowly, one to the other. Cooling, such as by an ice bath, can be used to maintain the reaction temperature within a suitable range, such as about 0°C to less than 50°C, 0°C to 20°C or 0°C to 10°C. After the addition is complete, the reaction is allowed to proceed until the reaction is substantially complete, and it may be allowed to warm to room temperature to promote the reaction. Optionally, additional nitrating agents or mixtures of nitrating agents may be added to promote the reaction to completion. The reaction is then quenched, such as by adding to water and / or ice, and the product is separated or extracted from an aqueous solution, and purified if necessary. Purification techniques suitable for purifying the product from any reaction disclosed herein include, but are not limited to, crystallization, distillation and / or chromatography.

[0114] Continuing with Scheme 2, compound 14 is then reacted with compound 16 to form compound 18. Compound 16 contains the desired R 1 moiety and a suitable leaving group LG. Suitable leaving groups include those that will act as a leaving group to facilitate the removal of R 1 Partially added to any group of compound 14. Suitable leaving groups include but are not limited to halogen (usually bromine, chlorine or iodine) and tosylate or mesylate groups. In a suitable solvent and usually in the presence of a base, compound 14 is reacted with compound 16. Suitable solvents include any solvents that promote the reaction, such as aprotic solvents. Suitable solvents include but are not limited to DMF, THF, DMSO, acetonitrile, chlorinated solvents (such as dichloromethane and chloroform), DMA, dioxane, N-methylpyrrolidone or a combination thereof. Suitable bases include any bases that will promote the reaction, such as hydrides (usually sodium hydride) or carbonates (such as potassium carbonate, sodium carbonate or cesium carbonate). As needed, reactants can be heated to such as 50°C, 100°C or higher, or the reaction can be carried out at room temperature. Then compound 18 is separated from the reaction mixture and purified if necessary.

[0115] Compound 18 is then reacted with a reducing agent 20 suitable for reducing the nitro moiety to an amine. Suitable reducing agents include, but are not limited to: hydrogen in the presence of a catalyst such as a palladium catalyst; borohydrides, such as sodium borohydride, optionally in the presence of a catalyst such as a nickel catalyst; zinc metal in acetic acid; or iron powder in water or water and acid. In certain embodiments, hydrogen is used in the presence of a palladium carbon catalyst and in a suitable solvent such as ethyl acetate or methanol. In some embodiments, a combination of reducing agents and / or techniques is used. For example, a first method comprising a first reducing agent and / or technique may first be reduced, but a mixture of products is produced. The first method may be repeated, and / or a second method comprising a second reducing agent and / or technique may be performed. Once the reaction is complete, as indicated by analytical techniques such as LC-MS, TLC or HPLC, the product compound 22 is separated and purified if necessary.

[0116] The third step of an exemplary reaction sequence is provided below according to Scheme 3:

[0117]

[0118] Compound 22 is reacted with carboxylic acid 24 to form compound 26. Carboxylic acid 24 is activated by any suitable method and then reacted with the amine on compound 22. Suitable activation methods include, but are not limited to, treatment with thionyl chloride to form an acid chloride; treatment with 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU) and a base such as diisopropylethylamine (DIPEA); treatment with carbonyldiimidazole (CDI); or treatment with a carbodiimide, such as dicyclohexylcarbodiimide (DCC) or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC).

[0119] Compound 26 is then coupled with compound 28 to form compound 30 using any coupling reaction suitable for forming a bond between the two rings. Boronic acid coupling is shown in the above examples, where the leaving group LG on compound 26 is typically bromine or iodine. Other suitable coupling functional groups include trialkyltin or borate esters. The coupling reaction is typically carried out in the presence of a suitable catalyst. For boronic acid coupling, the catalyst is typically a palladium catalyst such as PdCl 2 (dppf) 2 、Pd[P(Ph) 3 ] 2 Cl 2, palladium acetate and triphenylphosphine or tetrakis (triphenylphosphine) palladium (0).Reaction is carried out in the presence of a base (such as sodium carbonate, potassium carbonate or cesium carbonate), and in a suitable solvent or solvent mixture (such as dioxane, dioxane / water or DME / ethanol / water).Reaction can be heated at a suitable temperature (such as 50 ℃ to 125 ℃, usually about 100 ℃), and / or stir a suitable period of time (such as 1 hour to 3 days, 6 hours to 24 hours or 12 hours to 18 hours) to promote the reaction to complete.Then compound 30 is separated from the reaction mixture and purified by a suitable technology.

[0120] An alternative exemplary synthesis may include the following first reaction step according to Scheme 4:

[0121]

[0122] Compound 32 is nitrated to form compound 36 using a suitable nitrating agent or a mixture of reagents 34. Suitable nitrating conditions include reacting compound 32 with nitric acid (such as fuming nitric acid) (optionally in the presence of sulfuric acid). Typically, compound 32 and nitric acid are added slowly, one to the other. Cooling, such as by an ice bath, can be used to maintain the reaction temperature within a suitable range, such as about 0°C to less than 50°C, 0°C to 20°C or from 0°C to 10°C. After the addition is complete, the reaction is allowed to proceed until the reaction is substantially complete, and it may be allowed to warm to room temperature to promote the reaction. Optionally, additional nitrating agents or mixtures of nitrating agents may be added to promote the reaction to completion. The reaction is then quenched, such as by adding water and / or ice, and the product is separated or extracted from an aqueous solution, and purified if necessary. Purification techniques suitable for purifying the product from any reaction disclosed herein include, but are not limited to, crystallization, distillation and / or chromatography.

[0123] Continuing with reference to scheme 4, compound 36 is then reacted with compound 38 to form compound 40. Compound 38 comprises a desired ring (such as cyclobutyl, cyclopentyl or cyclohexyl ring) and a suitable leaving group LG. Suitable leaving groups include any group that will serve as a leaving group to promote the addition of a ring to compound 36. Suitable leaving groups include but are not limited to halogen (typically bromine, chlorine or iodine) and tosylate or mesylate groups. In a suitable solvent and typically in the presence of a base, compound 36 is reacted with compound 38. Suitable solvents include any solvent that promotes the reaction, such as aprotic solvents. Suitable solvents include but are not limited to DMF, THF, DMSO, acetonitrile, chlorinated solvents (such as dichloromethane and chloroform), DMA, dioxane, N-methylpyrrolidone or a combination thereof. Suitable bases include any base that will promote the reaction, such as hydride (typically sodium hydride) or carbonate (such as potassium carbonate, sodium carbonate or cesium carbonate). If desired, the reactants may be heated to temperatures such as 50°C, 100°C or higher, or the reaction may be carried out at room temperature. Compound 40 is then isolated from the reaction mixture and purified if desired.

[0124] Compound 40 is then reacted with a reducing agent 42 suitable for reducing the carbonyl moiety to a hydroxyl group. Suitable reducing agents include, but are not limited to, sodium borohydride, diisobutylaluminum hydride or lithium aluminum hydride. The reaction is carried out in a solvent suitable for promoting the reaction (such as an alcohol, particularly methanol or ethanol; THF; or diethyl ether). The reactants may be heated to, for example, 50°C, 100°C or higher, and cooled to, for example, less than 20°C, less than 10°C, less than 0°C or lower, as required, or the reaction may be carried out at room temperature. Once the reaction is complete, as indicated by analytical techniques (such as LC-MS, TLC or HPLC), the product compound 44 is separated and purified if necessary by a suitable technique (such as column chromatography).

[0125] Optionally, compound 44 can be reacted with compound 46 to form compound 48. Compound 46 contains the desired R x moiety and a suitable leaving group LG. Suitable leaving groups include those that will act as a leaving group to facilitate the removal of R xPartially added to any group of compound 44. Suitable leaving groups include, but are not limited to, halogens (usually bromine, chlorine or iodine) and tosylate or mesylate groups. In a suitable solvent and usually in the presence of a base or other reagent or a reagent that promotes the reaction, compound 44 is reacted with compound 46. Suitable solvents include any solvent that promotes the reaction, such as an aprotic solvent. Suitable solvents include, but are not limited to, DMF, THF, DMSO, acetonitrile, chlorinated solvents (such as dichloromethane and chloroform), DMA, dioxane, N-methylpyrrolidone or a combination thereof. Suitable bases or reagents that promote the reaction include, but are not limited to, silver trifluoromethanesulfonate, 2,6-di-tert-butylpyridine, sodium hydride or a combination thereof. Typically, compound 46 is slowly combined with the reactant. Cooling, such as by an ice bath, can be used to maintain the reaction temperature within a suitable range, such as about 0°C to less than 50°C, 0°C to 20°C or 0°C to 10°C. After the addition is complete, the reaction is allowed to proceed until the reaction is substantially complete and may be allowed to warm to room temperature to facilitate the reaction, or the reactants may be heated to, for example, 50° C., 100° C. or higher to facilitate the reaction. Once the reaction is complete, as indicated by analytical techniques such as LC-MS, TLC or HPLC, the product compound 48 is isolated by a suitable technique such as column chromatography and purified if necessary.

[0126] Alternatively, compound 40 can be prepared according to the exemplary synthetic route of Scheme 5:

[0127]

[0128] About scheme 5, compound 36 is reacted with compound 50 to form compound 52.Compound 50 comprises required ring (such as cyclobutyl, cyclopentyl or cyclohexyl ring), suitable leaving group (LG) and protected carbonyl moiety (such as acetal or ketal).In the above example, cyclic ketal moiety is shown.Suitable leaving group includes any group that will serve as leaving group to promote the ring to be added to compound 36, and includes but is not limited to halogen (usually bromine, chlorine or iodine) and tosylate or mesylate group.In suitable solvent and usually in the presence of base, compound 36 is reacted with compound 50.Suitable solvent includes any solvent that promotes reaction, such as aprotic solvent.Suitable solvent includes but is not limited to DMF, THF, DMSO, acetonitrile, chlorinated solvent (such as dichloromethane and chloroform), DMA, dioxane, N-methylpyrrolidone or its combination.Suitable alkali includes any alkali that will promote reaction, such as hydride (usually sodium hydride) or carbonate (such as potassium carbonate, sodium carbonate or cesium carbonate). If desired, the reactants may be heated to temperatures such as 50°C, 100°C or higher, or the reaction may be carried out at room temperature. Compound 52 is then isolated from the reaction mixture and purified if necessary by a suitable technique such as column chromatography.

[0129] Compound 52 is then reacted with a suitable reagent 54 to form compound 40. Reagent 54 may be any reagent suitable for removing protecting groups and / or forming a carbonyl moiety. In the exemplary synthesis shown in scheme 5, the protecting group is a cyclic ketal, and suitable reagent 54 includes but is not limited to pyridinium toluenesulfonate (PPTS), p-toluenesulfonic acid, hydrochloric acid or acetic acid. The reaction is carried out in a solvent or a mixture of solvents (such as acetone, THF, acetic acid, water or a combination thereof) suitable for promoting the reaction. The reactant may be heated to, such as 50° C., 100° C. or higher, or, as required, under reflux, or the reaction may be carried out at room temperature. Compound 40 is then separated from the reaction mixture and purified by a suitable technique (such as column chromatography) if necessary.

[0130] Step 2 of an exemplary reaction sequence is provided below according to Scheme 6:

[0131]

[0132] Compound 48 is then reacted with a reducing agent 56 suitable for reducing the nitro moiety to an amine. In certain embodiments, where the desired product compound contains a hydroxyl moiety, compound 44 may be used in place of compound 48. Suitable reducing agents include, but are not limited to: hydrogen in the presence of a catalyst such as a palladium catalyst; borohydrides, such as sodium borohydride, optionally in the presence of a catalyst such as a nickel catalyst; zinc metal in acetic acid; or iron powder in water or in water and acid. In certain embodiments, hydrogen is used in the presence of a palladium carbon catalyst and in a suitable solvent such as ethyl acetate or methanol. In some embodiments, a combination of reducing agents and / or techniques is used. For example, a first method including a first reducing agent and / or technique may be first reduced, but a mixture of products is produced. The first method may be repeated, and / or a second method including a second reducing agent and / or technique may be performed. Once the reaction is complete, product compound 58 is separated and purified if necessary, as indicated by analytical techniques such as LC-MS, TLC, or HPLC.

[0133] Compound 58 is reacted with carboxylic acid 60 to form compound 62. Carboxylic acid 60 is activated by any suitable method and then reacted with the amine on compound 58. Suitable activation methods include, but are not limited to: forming an acid chloride by treatment with thionyl chloride; treating with 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU) and a base such as diisopropylethylamine (DIPEA); treating with carbonyldiimidazole (CDI); or treating with a carbodiimide such as dicyclohexylcarbodiimide (DCC) or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC).

[0134] Compound 62 is then coupled with compound 64 to form compound 66 using any coupling reaction suitable for forming a bond between the two rings. In the above example, a boronate coupling is shown, where the leaving group LG on compound 62 is typically bromine or iodine. Other suitable coupling functional groups include trialkyltin or boronic acid. The coupling reaction is typically carried out in the presence of a suitable catalyst. For boronate or boronic acid coupling, the catalyst is typically a palladium catalyst such as PdCl 2 (dppf) 2 、Pd[P(Ph) 3 ] 2 Cl 2, palladium acetate and triphenylphosphine or tetrakis (triphenylphosphine) palladium (0).Reaction is carried out in the presence of a base (such as sodium carbonate, potassium carbonate or cesium carbonate), and in a suitable solvent or solvent mixture (such as dioxane, dioxane / water or DME / ethanol / water) is carried out.Reactant can be heated at a suitable temperature (such as 50 ℃ to 125 ℃, usually about 100 ℃), and / or stir a suitable period of time (such as 1 hour to 3 days, 6 hours to 24 hours or 12 hours to 18 hours) to promote reaction to complete.Then compound 66 is separated from the reaction mixture and purified by a suitable technology.

[0135] Certain embodiments may comprise a phosphate moiety. Scheme 7 provides an exemplary synthesis of certain such embodiments:

[0136]

[0137] Compound 68 is reacted with compound 70 to form compound 72. Compound 70 contains the desired R y moiety and a suitable leaving group LG. Typical R y Moieties include, but are not limited to, aliphatic (such as alkyl, typically methyl, ethyl, propyl, isopropyl, or tert-butyl); aryl; heteroaliphatic; or heterocyclic. y Parts may be the same or different. Suitable leaving groups include, but are not limited to, halogens (typically bromine, chlorine or iodine) and tosylate or mesylate groups. In a suitable solvent and usually in the presence of a base, compound 68 is reacted with compound 70. Suitable solvents include any solvent that promotes the reaction, such as aprotic solvents. Suitable solvents include, but are not limited to, DMF, THF, DMSO, acetonitrile, chlorinated solvents (such as dichloromethane and chloroform), DMA, dioxane, N-methylpyrrolidone or a combination thereof. Suitable bases include any base that will promote the reaction, such as hydrides (typically sodium hydride) or carbonates (such as potassium carbonate, sodium carbonate or cesium carbonate). As needed, the reactants can be heated to, such as 50°C, 100°C or higher, or the reaction can be carried out at room temperature. Then compound 72 is separated from the reaction mixture and purified if necessary.

[0138] Compound 72 is then reacted with compound 74 to form compound 76. Compound 74 may be any compound suitable for forming an acid moiety in compound 76. Compound 74 may be an acidic reagent such as trifluoroacetic acid, hydrochloric acid or hydrobromic acid, or it may be an alkaline reagent such as sodium hydroxide, lithium hydroxide or potassium hydroxide. Suitable solvents include, but are not limited to, chlorinated solvents (such as dichloromethane and chloroform), alcohols (such as methanol and ethanol), water or a combination thereof. The reactants may be heated to, for example, 50°C, 100°C or higher, and cooled to, for example, less than 20°C, less than 10°C, less than 0°C or lower, as required, or the reaction may be carried out at room temperature. As indicated by analytical techniques (such as LC-MS, TLC or HPLC), once the reaction is complete, the product compound 76 may be separated and purified if necessary in a suitable solvent or solvent system by a suitable technique (such as by agitation, such as by stirring or ultrasonic treatment). Suitable solvents or solvent systems include, but are not limited to, acetone / water, acetone, diethyl ether or alcohol / water.

[0139] Compound 76 is then reacted with compound 78 to form salt compound 80. Compound 78 can be any compound that will provide a suitable counterion CA for salt compound 80, such as calcium hydroxide, sodium hydroxide, potassium hydroxide, lithium hydroxide, ammonium, trimethylamine, tris(hydroxymethyl)aminomethane, or an amino acid such as lysine or arginine. One of ordinary skill in the art will appreciate that if the counterion CA has a single positive charge (such as in Na + , K + , Li + or NH 4 + ), then compound 80 will contain two CA ions, whereas if the counter ion CA has two positive charges (such as in CA 2+ form), then compound 80 will contain a CA ion.

[0140] IV. Combinations of Therapeutic Agents

[0141] The compounds of the present disclosure may be used alone, in combination with each other, in separate pharmaceutical compositions, together in a single composition, or as an adjunct to or in combination with other established therapies. One or more compounds may be administered once, or more likely multiple times. On the other hand, the compounds of the present disclosure may be used in combination with other therapeutic agents useful for the condition or illness being treated. These compounds and / or agents may be administered simultaneously, sequentially in any order, by the same route of administration or by different routes. For sequential administration, one or more compounds and / or one or more agents may be administered so that the effective time period of at least one compound and / or agent overlaps with the effective time period of at least one other compound and / or agent. In an exemplary embodiment of a combination comprising four components, the effective time period of the first component administered may overlap with the effective time period of the second, third, and fourth components, but the effective time periods of the second, third, and fourth components independently may or may not overlap with each other. In another exemplary embodiment of a combination comprising four components, the effective time period of the first component administered overlaps with the effective time period of the second component, but does not overlap with the effective time period of the third or fourth component; the effective time period of the second component overlaps with the effective time period of the first and third components; and the effective time period of the fourth component overlaps only with the effective time period of the third component. In some embodiments, the effective time periods of all compounds and / or agents overlap with each other.

[0142] In some embodiments, the disclosed compounds are administered with another therapeutic agent such as an analgesic, antibiotic, anticoagulant, antibody, anti-inflammatory agent, immunosuppressant, guanylate cyclase C agonist, intestinal secretagogue, antiviral agent, anticancer agent, antifungal agent, or a combination thereof. In certain embodiments, the second therapeutic agent is an anti-inflammatory agent, an immunosuppressant, and / or may be a steroid.

[0143] The anti-inflammatory agent may be a steroid or a non-steroidal anti-inflammatory agent. In certain embodiments, the non-steroidal anti-inflammatory agent is selected from aminosalicylate, cyclooxygenase inhibitors, diclofenac, etodolac, famotidine, fenoprofen, flurbiprofen, ketoprofen, ketorolac, ibuprofen, indomethacin, meclofenamic acid, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, piroxicam, salsalate, sulindac, tolmetin or a combination thereof. In some embodiments, the immunosuppressant is mercaptopurine, corticosteroids, alkylating agents, calcineurin inhibitors, inosine monophosphate dehydrogenase inhibitors, antilymphocyte globulin, antithymocyte globulin, anti-T cell antibodies or a combination thereof. In one embodiment, the antibody is infliximab.

[0144] In some embodiments, the compounds of the present invention can be used together with anticancer agents or cytotoxic agents. Various classes of anticancer agents and antitumor compounds include, but are not limited to, alkylating agents, antimetabolites, BCL-2 inhibitors, vinca alkaloids, taxanes, antibiotics, enzymes, cytokines, platinum coordination complexes, proteasome inhibitors, substituted ureases, kinase inhibitors, hormones and hormone antagonists, and hypomethylating agents (e.g., DNMT inhibitors, such as azacitidine and decitabine). Exemplary alkylating agents include, but are not limited to, nitrogen mustard, cyclophosphamide, ifosfamide, melphalan, chlorambucil, ethyleneimine, methyl melamine, alkyl sulfonates (e.g., busulfan) and carmustine. Exemplary antimetabolites include, for example, but not limited to, folic acid analog methotrexate; pyridine analogs fluorouracil, cytosine arabinoside; purine analogs mercaptopurine, thioguanine and thioazapurine. Exemplary vinca alkaloids include, by way of example but not limited to, vinblastine, vincristine, paclitaxel, and colchicine. Exemplary antibiotics include, by way of example but not limited to, actinomycin D, daunomycin, and bleomycin. Exemplary enzymes effective as antitumor agents include L-asparaginase. Exemplary coordination compounds include, by way of example but not limited to, cisplatin and carboplatin. Exemplary hormones and hormone-related compounds include, by way of example but not limited to, the adrenocortical steroids prednisone and dexamethasone; the aromatase inhibitors aminoglutethimide, formestane, and anastrozole; the progesterone compounds hydroxyprogesterone hexyl ester, medroxyprogesterone; and the anti-estrogen compound tamoxifen.

[0145] These and other useful anticancer compounds are described in the Merck Index, 13th Edition (O'Neil MJ et al., eds.) Merck Publishing Group (2001); and in Goodman and Gilman's The Pharmacological Basis of Therapeutics, 12th Edition, Brunton LL, ed., Chapters 60-63, McGraw Hill, (2011), both of which are incorporated herein by reference.

[0146] Among the CTLA4 antibodies that can be used in combination with the inhibitors disclosed herein are Ipilimumab for sale.

[0147] Other chemotherapeutic agents for combination include immuno-oncology agents, such as checkpoint pathway inhibitors, for example, PD-1 inhibitors (such as nivolumab and lambrolizumab) and PD-L1 inhibitors (such as pembrolizumab, MEDI-4736 and MPDL3280A / RG7446). Additional checkpoint inhibitors for combination with the compounds disclosed herein include anti-LAG-3 agents, such as BMS-986016 (MDX-1408).

[0148] Additional chemotherapeutic agents for combination with the inhibitors of the present disclosure include anti-SLAMF7 agents such as the humanized monoclonal antibody elotuzumab (BMS-901608), anti-KIR agents such as the anti-KIR monoclonal antibody lirilumab (BMS-986015), and anti-CD137 agents such as the fully human monoclonal antibody urelumab (BMS-663513).

[0149] Additional anti-proliferative compounds useful in combination with the compounds of the present disclosure include, by way of example but not limitation, antibodies directed against growth factor receptors (eg, anti-Her2); and cytokines, such as interferon-α and -γ, interleukin-2, and GM-CSF.

[0150] Additional chemotherapeutic agents useful in combination with the compounds of the invention include proteasome inhibitors such as bortezomib, carfilzomib, marizomib, and the like.

[0151] Examples of kinase inhibitors useful in combination with the compounds of the present disclosure, particularly in the treatment of malignancies, include: Btk inhibitors such as ibrutinib; CDK inhibitors such as palbociclib; EGFR inhibitors such as afatinib, erlotinib, gefitinib, lapatinib, osimertinib, and vandetinib; Mek inhibitors such as trametinib; Raf inhibitors such as dabrafenib, sorafenib, and vemurafenib; VEGFR inhibitors such as axitinib, lenvatinib, nintedanib, pazopanib; BCR-Abl inhibitors such as bosutinib, dasatinib, imatinib, and nilotinib; Syk inhibitors such as fostamatinib; and JAK inhibitors such as ruxolitinib.

[0152] In other embodiments, the second therapeutic agent may be selected from any of the following:

[0153] Analgesics-morphine, fentanyl, hydromorphone, oxycodone, codeine, acetaminophen, hydrocodone, buprenorphine, tramadol, venlafaxine, flupirtine, meperidine, pentazocine, dextromorphone, dipipanone;

[0154] Antibiotics - aminoglycosides (for example, amikacin, gentamicin, kanamycin, neomycin, netilmicin, tobramycin, and paromomycin), carbapenems (for example, ertapenem, doripenem, imipenem, cilastatin, and meropenem), cephalosporins (for example, cefdroxil, cefazolin, cephalothin, cephalexin, cefaclor, cefamandole, cefoxitin, cefprozil, cefuroxime , cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten, cefazolin, ceftriaxone, cefepime, and cefobiprole), glycopeptides (e.g., teicoplanin, vancomycin, and telavancin), lincosamides (e.g., clindamycin and lincomycin), lipopeptides (e.g., daptomycin), macrolides (adenosine disulfiram, daptomycin), and oxadiazine. erythromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, and spectinomycin), monobactams (e.g., aztreonam), nitrofurans (e.g., furazolidone and nitrofurantoin), penicillins (e.g., amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin G, penicillin V, piperacillin, temocillin, and ticarcillin), penicillin combinations (e.g., amoxicillin / clavulanic acid, ampicillin / sulbactam, piperacillin / tazobactam, and ticarcillin / clavulanic acid), polypeptides (e.g., bacitracin, colistin, and polymyxin B), quinolinones (e.g., ciprofloxacin, enoxacin, gatifloxacin, levofloxacin, lomefloxacin, moxifloxacin, sulfonamides (e.g., mafenide, sulfonamidochrysoidine, sulfacetamide, sulfadiazine, silver sulfadiazine, sulfamethoxazole, sulfamethoxazole, sulfanilimide, sulfasalazine, sulfisoxazole, trimethoprim, and trimethoprim-sulfamethoxazole), tetracyclines (e.g., demeclocycline, doxycycline, minocycline, oxytetracycline and tetracycline), antimycobacterial compounds (e.g., clofazimine, dapsone, capreomycin, cycloserine, ethambutol, ethionamide, isoniazid, pyrazinamide, rifampicin (rifampin), rifabutin, rifapentine, and streptomycin), and others (such as arsphenamine, chloramphenicol, fosfomycin, fusidic acid, linezolid, metronidazole, mupirocin, platenmycin, quinuprisin / dalfopristin, rifaximin, thiamphenicol, tigecycline, and tinidazole);

[0155] Antibodies - Anti-TNF-α antibodies, such as Remicade TM ), adalimumab, golimumab, certolizumab; anti-B cell antibodies, such as rituximab; anti-IL-6 antibodies, such as tocilizumab; anti-IL-1 antibodies, such as anakinra; anti-PD-1 and / or anti-PD-L1 antibodies, such as nivolumab, pembrolizumab, pidilizumab, BMS-936559, MPDL3280A, AMP-224, MEDI4736; ixekizumab, brodalumab ), ofatumumab, sirukumab, cliciximab, clazakiumab, fezakinumab, fletikumab, mavrilimumab, ocrelizumab, sarilumab, secukinumab, toralizumab, zalumab;

[0156] Anticoagulants - Warfarin TM ), acenaphthenocoumarol, phenprocoumon, schizotin, phenindione, heparin, fondaparinux sodium, idraparinux, rivaroxaban, apixaban, hirudin, lepirudin, bivalirudin, argatrobam, dabigatran, ximelagatran, batroxobin, fibrase;

[0157] Anti-inflammatory agents - steroids (e.g., budesonide), nonsteroidal anti-inflammatory agents (e.g., aminosalicylates (e.g., sulfasalazine, mesalazine, olsalazine, and balsalazide), cyclooxygenase inhibitors (COX-2 inhibitors such as rofecoxib, celecoxib), diclofenac, etodolac, famotidine, fenoprofen, flurbiprofen, ketoprofen, ketorolac, ibuprofen, indomethacin, meclofenamic acid, mefenamic acid, meloxicam, naproxen, naproxen, oxaprozin, piroxicam, salsalate, sulindac, tolmetin);

[0158] Immunosuppressants - mercaptopurines, corticosteroids (such as dexamethasone, hydrocortisone, prednisone, methylprednisolone and prednisolone), alkylating agents (such as cyclophosphamide), calcineurin inhibitors (such as cyclosporine, sirolimus and tacrolimus), inosine monophosphate dehydrogenase (IMPDH) inhibitors (such as mycophenolate mofetil, mycophenolate mofetil and azathioprine), and agents designed to suppress cellular immunity while leaving the recipient's humoral immune response intact, including various antibodies (e.g., antilymphocyte globulin (ALG), antithymocyte globulin (ATG), monoclonal anti-T-cell antibodies (OKT3)) and radiation. Azathioprine is currently available from Salix Pharmaceuticals, Inc. under the trade name Azasan; mercaptopurine is currently available from Gate Pharmaceuticals, Inc. under the trade name Purinethol; prednisone and prednisolone are currently available from Roxane Laboratories, Inc.; methylprednisolone is currently available from Pfizer; sirolimus (rapamycin) is currently available from Wyeth-Ayerst under the trade name Rapamune; tacrolimus is currently available from Fujisawa under the trade name Prograf; cyclosporine is currently available from Novartis under the trade name Sandimmune and Abbott under the trade name Gengraf; IMPDH inhibitors (such as mycophenolate mofetil and mycophenolic acid) are currently available from Roche under the trade name Cellcept and Novartis under the trade name Myfortic; azathioprine is currently available from Glaxo Smith Kline under the trade name Imuran; and antibodies are currently available from Ortho Orthoclone from Biotech, Simulect (basiliximab) from Novartis, and Zenapax (daclizumab) from Roch; and

[0159] A guanylate cyclase C receptor agonist or intestinal secretagogue, for example linaclotide sold under the name Linzess.

[0160] In certain embodiments, the second therapeutic agent is or comprises a steroid, such as a corticosteroid, including but not limited to glucocorticoids and / or mineralocorticoids.Steroids suitable for use in combination with the disclosed compounds include synthetic and non-synthetic glucocorticoids.Exemplary steroids (such as glucocorticoids) suitable for use in the disclosed methods include, but are not limited to, alclomethasone, algestrone, beclomethasone (e.g., beclomethasone dipropionate), betamethasone (e.g., betamethasone 17-valerate, betamethasone sodium acetate, betamethasone sodium phosphate, betamethasone valerate), budesonide, clobetasol (e.g., clobetasol propionate), clobetasone, clocortolone (e.g., clocortolone pivalate), cloprednisolone, corticosterone, cortisone, dexamethasone, deflazacort, desonide, deschlorodexamethasone, dexamethasone (e.g., dexamethasone 21-phosphate, dexamethasone acetate, dexamethasone sodium phosphate), diflorasone (e.g., diflorasone diacetate), diflucortolone, difluprednate, glycyrrhetinic acid, flurazepam, flucloroxolone, fludrocortisone (e.g., fludrocortisone acetate), flumethasone (e.g., flumethasone pivalate), flunisolide, fluocinolone (e.g., fluocinolone acetate acetonide), fluocinonide, fluocortidine, fluocortolone, fluorometholone (e.g., fluorometholone acetate), fluperone (e.g., fluperone acetate), fluprednidine, fluprednisolone, fluocortidine, fluticasone (e.g., fluticasone propionate), formocortal, clofloxacin, halobetasol, halometasone, halopednisone, hydrocortisone, hydrocortisone (e.g., hydrocortisone 21-butyrate, hydrocortisone acetonide, hydrocortisone acetate, hydrocortisone propylbutyrate, hydrocortisone butyrate, hydrocortisone cypionate, hydrocortisone hemisuccinate, hydrocortisone propylbutyrate, hydrocortisone sodium phosphate, hydrocortisone sodium succinate, hydrocortisone valerate), loteprednol etabonate, maprednione, medrysone, methylprednisone, methylprednisolone (methylprednisolone acetonide, methylprednisolone acetate, methylprednisolone hemisuccinate, methylprednisolone sodium succinate), mometasone (e.g., methylprednisolone furoate), paramethasone (e.g., paramethasone acetate), prednicarbate, prednisolone (e.g., prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisolone 21-hemisuccinate, prednisolone acetate; prednisolone farnesate, prednisolone hemisuccinate, prednisolone-21 (β-D-glucuronic acid), prednisolone meta-sulfobenzoate, prednisolone stilbene, prednisolone butyrate, prednisolone tetrahydrophthalate), prednisone, prednival, prednididine, rimexolone, tixocortolone, triamcinolone acetonide (e.g., triamcinolone acetate acetonide, triamcinolone benetonide, triamcinolone hexacetonide, triamcinolone acetonide 21-palmitate, triamcinolone diacetate), or any combination thereof.Additional information on steroids and their salts can be found, for example, in Remington's Pharmaceutical Sciences, A. Osol, ed., Mack Pub. Co., Easton, Pa. (16th ed. 1980).

[0161] In some instances, the steroid is a glucocorticoid and can be selected from cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone or a combination thereof. In a specific instance, the steroid is or comprises prednisone. In another specific instance, the steroid is or comprises dexamethasone.

[0162] These various agents may be used according to their standard or conventional dosages as indicated in the prescribing information of the commercially available form of the accompanying medication (see also, prescribing information in the 2006 edition of The Physician's Desk Reference), the disclosure of which is incorporated herein by reference.

[0163] V. Compositions Containing the Disclosed Compounds

[0164] The disclosed compounds can be used individually, in any combination and in combination with at least one second therapeutic agent or as its auxiliary. In addition, the disclosed one or more compounds and / or at least one second therapeutic agent can be used in combination with any suitable excipient, and the excipient can be used to form a composition applied to a subject. Excipients may be included in pharmaceutical compositions for various purposes, such as for diluting the composition delivered to the subject, for promoting the processing of preparations, for providing favorable material properties to preparations, for promoting dispersion from delivery devices, for stabilizing preparations (e.g., antioxidants or buffers), for providing satisfactory or delicious taste or consistency to preparations, etc. One or more pharmaceutically acceptable excipients may include one or more pharmaceutically acceptable carriers and / or pharmaceutically acceptable adjuvants. Exemplary excipients include, but are not limited to, monosaccharides, disaccharides, and polysaccharides, sugar alcohols and other polyols, such as lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch, or combinations thereof; surfactants, such as sorbitol, diphosphatidylcholine, and lecithin; bulking agents; buffers, such as phosphate and citric acid buffers; antiadherents, such as magnesium stearate; binders, such as sugars (including disaccharides, such as sucrose and lactose), polysaccharides (such as starch, cellulose, microcrystalline cellulose, cellulose esters (such as hydroxypropyl cellulose)), gelatin, synthetic polymers (such as polyvinyl pyrrolidone, polyalkylene glycols); coatings (such as cellulose ethers, including hydroxypropyl methylcellulose, shellac, corn protein zein, and gelatin); release aids (such as enteric coatings); disintegrants (such as crosslinking agents); povidone, cross-linked sodium carboxymethylcellulose, and sodium starch glycolate); fillers (such as dibasic calcium phosphate, vegetable fats and oils, lactose, sucrose, glucose, mannitol, sorbitol, calcium carbonate, and magnesium stearate); flavoring and sweetening agents (such as mint, cherry, anise, peach, apricot, or licorice, raspberry, and vanilla); lubricants (such as minerals, exemplified by talc or silicon dioxide; fats, exemplified by vegetable stearin, magnesium stearate, or stearic acid); preservatives (such as antioxidants, exemplified by vitamin A, vitamin E, vitamin C, retinyl palmitate, and selenium, amino acids (exemplified by cysteine ​​and methionine), citric acid and sodium citrate, parabens (exemplified by methylparaben and propylparaben)); colorants; compression aids; emulsifiers; encapsulating agents; gelatin; granulating agents; and combinations thereof.

[0165] VI. How to use

[0166] A. Disease / condition

[0167] The disclosed compounds and combinations and / or compositions thereof can be used to improve, treat and / or prevent various diseases, illnesses and / or conditions. In certain embodiments, the disclosed compounds, combinations of disclosed compounds or compositions thereof can be used to treat conditions in which inhibition of interleukin-1 receptor-associated kinase (IRAK) pathways is therapeutically useful. In some embodiments, the compound directly inhibits IRAK proteins, such as IRAK1, IRAK2, IRAK3 and / or IRAK4. In certain embodiments, the disclosed compounds can be used to treat, prevent and / or improve autoimmune diseases, inflammatory conditions, cardiovascular diseases, neurological conditions, neurodegenerative conditions, allergic conditions, asthma, pancreatitis, multiple organ failure, kidney disease, platelet aggregation, cancer, transplantation, sperm motility, erythrocyte deficiency, transplant rejection, lung injury, respiratory diseases, ischemic conditions and bacterial and viral infections.

[0168] In some embodiments, the disclosed compounds, combinations of disclosed compounds, or compositions thereof can be used to treat or prevent allergic diseases, amyotrophic lateral sclerosis (ALS), systemic lupus erythematosus, rheumatoid arthritis, type I diabetes, inflammatory bowel disease, biliary cirrhosis, uveitis, multiple sclerosis, Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, psoriasis, autoimmune myositis, Wegener's granulomatosis, ichthyosis, Graves' ophthalmopathy, or asthma.

[0169] The disclosed compounds, combinations of disclosed compounds, or compositions thereof may also be used to ameliorate, treat, and / or prevent immunoregulatory disorders associated with bone marrow or organ transplant rejection or graft-versus-host disease.Examples of inflammatory or immunomodulatory disorders that may be treated with the compounds of the invention include, but are not limited to, organ or tissue transplantation, graft-versus-host disease acquired by transplantation, autoimmune syndromes (including rheumatoid arthritis), lupus (including systemic lupus erythematosus), Hashimoto's thyroiditis, multiple sclerosis, systemic sclerosis, myasthenia gravis, type I diabetes, uveitis, posterior uveitis, allergic encephalomyelitis, glomerulonephritis, post-infectious autoimmune diseases (including rheumatic fever and post-infectious glomerulonephritis), inflammatory and hyperproliferative skin diseases, psoriasis, atopic dermatitis , contact dermatitis, eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermolysis bullosa, urticaria, angioedema, vasculitis, erythema, cutaneous eosinophilia, lupus erythematosus, acne, alopecia areata, keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic keratitis, keratoconus, corneal epithelial dystrophy, corneal leukoplakia, ocular pemphigus, Mooren's ulcer, scleritis, Graves' ophthalmopathy, Vogt-Koyanagi-Harada syndrome syndrome), sarcoidosis, pollen allergy, reversible obstructive airway disease, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma, dust asthma, chronic or deep-seated asthma, advanced asthma and airway hyperresponsiveness, bronchitis, gastric ulcer, vascular damage caused by ischemic disease and thrombosis, ischemic bowel disease, inflammatory bowel disease, necrotizing enterocolitis, intestinal lesions associated with thermal burns, celiac disease, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease, ulcerative colitis, migraine, rhinitis, eczema, interstitial nephritis, Goodpasture's syndrome, hemolytic uremic syndrome, diabetic nephropathy, polymyositis, Guillain-Barre syndrome, Meniere's disease, polyneuritis, polyneuritis, mononeuritis, radiculopathy, hyperthyroidism, Basedow's disease disease), pure red cell aplasia, aplastic anemia, hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia, erythropoiesis, osteoporosis, sarcoidosis, fibrosing lung, idiopathic interstitial pneumonia, dermatomyositis, vitiligo vulgaris, ichthyosis vulgaris, photoallergic sensitivity, cutaneous T-cell lymphoma, chronic lymphocytic leukemia, arteriosclerosis, atherosclerosis, aortitis syndrome, polyarteritis nodosa, cardiomyopathy, scleroderma, Wegener's granulomatosis, Sjögren's syndrome (. syndrome), obesity, eosinophilic fasciitis, gingival lesions, periodontal ligament lesions, alveolar bone lesions, substantia ossea dentis, glomerulonephritis, male pattern baldness or senile hair loss by preventing hair loss or providing hair generation and / or promoting hair generation and hair growth, muscular dystrophy, pyoderma and Sezary's syndrome, Addison's disease, ischemia-reperfusion injury after organ preservation, transplantation or ischemic disease, endotoxin shock, pseudomembranous colitis, colitis caused by drugs or radiation, ischemic acute renal insufficiency, chronic renal insufficiency, toxic diseases caused by pulmonary oxygen or drugs, lung cancer, emphysema, cataract, siderosis, retinitis pigmentosa, age-related macular degeneration, vitreous scars, corneal alkali burns, erythema polymorphum, linear IgA bullous dermatitis and cement dermatitis, gingivitis, periodontitis, sepsis, pancreatitis, diseases caused by environmental pollution, aging, carcinogenesis, cancer Metastasis and hypotension sickness, diseases caused by histamine or leukotriene-C4 release, Behcet's disease, autoimmune hepatitis, primary biliary cirrhosis, sclerosing cholangitis, partial hepatectomy, acute liver necrosis, necrosis caused by toxins, viral hepatitis, shock or hypoxia, viral hepatitis B, non-A / non-B hepatitis, cirrhosis, alcoholic liver disease (including alcoholic cirrhosis), non-alcoholic steatohepatitis (NASH), liver failure, fulminant liver failure, late-onset liver failure, "acute-on-chronic" liver failure, enhanced chemotherapy effects, cytomegalovirus infection, HCMV infection, AIDS, cancer, Alzheimer's disease, Parkinson's disease, trauma, chronic bacterial infection, palmoplantar pustulosis, hidradenitis suppurativa, cytokine release syndrome (CRS), acute respiratory distress syndrome (ARDS), acute kidney injury (AKI), renal insufficiency or thrombosis.

[0170] In some embodiments, the disease or condition is hidradenitis suppurativa or a lymphoid neoplasm selected from the group consisting of myeloproliferative neoplasms (MPNs) excluding polycythemia vera, myeloid / lymphoid neoplasms with PDGFRA rearrangement, myeloid / lymphoid neoplasms with PDGFRB rearrangement, myeloid / lymphoid neoplasms with FGFR1 rearrangement, myeloid / lymphoid neoplasms with PCM1-JAK2, myelodysplastic / myeloproliferative neoplasms (MDS / MPNs), myeloid sarcoma, myeloproliferation associated with Down syndrome, blastic plasmacytoid dendritic cell neoplasm, B lymphoblastic leukemia / lymphoma; and / or T lymphoblastic leukemia / lymphoma. In some embodiments, the lymphoid neoplasm is a myeloproliferative neoplasm selected from the group consisting of chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), primary myelofibrosis (PMF), essential thrombocythemia, chronic eosinophilic leukemia, or a combination thereof. In other embodiments, the lymphoid tumor is a myelodysplastic / myeloproliferative neoplasm selected from the group consisting of chronic myelomonocytic leukemia, atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia (JMML), MDS / MPN with ring sideroblasts and thrombocythemia (MDS / MPN-RS-T), or a combination thereof.

[0171] In certain embodiments, the compounds of the invention are useful for treating neurological pain, including neuropathic pain and pain resulting from inflammation.

[0172] In certain embodiments, the disclosed compounds, combinations of disclosed compounds, or compositions thereof are useful for treating and / or preventing rheumatoid arthritis, psoriatic arthritis, osteoarthritis, systemic lupus erythematosus, lupus nephritis, ankylosing spondylitis, osteoporosis, systemic sclerosis, multiple sclerosis, psoriasis (particularly pustular psoriasis), type I diabetes, type II diabetes, inflammatory bowel disease (Crohn's disease and ulcerative colitis), hyperimmunoglobulinemia and periodic fever syndromes, cryopyrin-associated periodic syndromes, Schnitzler's syndrome, systemic juvenile idiopathic arthritis, adult'sonset Still's disease, gout, gouty attacks, pseudogout, sapho syndrome, Castleman's disease, sepsis, stroke, atherosclerosis, celiac disease, DIRA (deficiency of 11-1 receptor antagonists), Alzheimer's disease, or Parkinson's disease.

[0173] Proliferative diseases that can be treated by the disclosed compounds, combinations of disclosed compounds, or compositions thereof include benign or malignant tumors, solid tumors, brain cancer, kidney cancer, liver cancer, adrenal cancer, bladder cancer, breast cancer, stomach cancer, gastric tumors, ovarian cancer, colon cancer, rectal cancer, prostate cancer, pancreatic cancer, lung cancer, vaginal cancer, cervical cancer, testicular cancer, genitourinary tract cancer, esophageal cancer, laryngeal cancer, skin cancer, bone cancer or thyroid cancer, sarcoma, glioblastoma, neuroblastoma, multiple myeloma, gastrointestinal cancer (especially is colon cancer or colorectal adenoma), head and neck tumors, epidermal hyperplasia, psoriasis, prostatic hyperplasia, neoplasia, epithelial neoplasia, adenoma, adenocarcinoma, keratoacanthoma, squamous cell carcinoma, large cell carcinoma, non-small cell lung cancer, lymphoma, Hodgkin and non-Hodgkin, breast cancer, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, IL-1 driven disorders, MyD88 driven disorders (such as ABC diffuse large B-cell lymphoma (DLBCL), Waldenstrom's macroglobulinemia ( macroglobulinemia), Hodgkin's lymphoma, primary cutaneous T-lymphocytic neoplasm or chronic lymphocytic leukemia), smoldering or indolent multiple myeloma or hematological malignancies (including leukemia, acute myeloid leukemia (AML), DLBCL, ABC DLBCL, chronic lymphocytic leukemia (CLL), chronic lymphocytic lymphoma, primary effusion lymphoma, Burkitt's lymphoma / leukemia, acute lymphocytic leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, myelodysplastic syndrome (MDS), myelofibrosis, polycythemia vera, Kaposi's sarcoma, Waldenstrom's macroglobulinemia (WM), splenic marginal zone lymphoma, multiple myeloma, plasmacytoma, intravascular large B-cell lymphoma). In particular, the compounds of the present disclosure may be used to treat drug-resistant malignancies, such as those resistant to the JAK inhibitor ibrutinib, including ibrutinib-resistant hematological malignancies, such as ibrutinib-resistant CLL and ibrutinib-resistant Waldenstrom's macroglobulinemia.

[0174] Examples of allergic conditions that can be treated using the disclosed compounds, combinations of disclosed compounds, or compositions thereof include, but are not limited to, asthma (e.g., atopic asthma, allergic asthma, atopic bronchial IgE-mediated asthma, non-atopic asthma, bronchial asthma, non-allergic asthma, idiopathic asthma, true asthma, intrinsic asthma caused by pathophysiological disorders, idiopathic asthma of unknown or inapparent cause, emphysematous asthma, exercise-induced asthma, emotion-induced asthma, exogenous asthma caused by environmental factors, Asthma, cold-air-induced asthma, occupational asthma, infectious asthma caused by or associated with bacterial, fungal, protozoal, or viral infection, early asthma, asthma-infant syndrome, bronchitis, cough variant asthma, or drug-induced asthma), allergic bronchopulmonary aspergillosis (ABPA), allergic rhinitis, perennial allergic rhinitis, perennial rhinitis, vasomotor rhinitis, postnasal drip, suppurative or non-purulent sinusitis, acute or chronic sinusitis, and ethmoid, frontal, maxillary, or sphenoid sinusitis.

[0175] As another example, rheumatoid arthritis (RA) usually causes swelling, pain, loss of motion and tenderness of systemic target joints. RA is characterized by a chronic inflammatory synovium of dense lymphocytes. Usually a cell layer thick synovium becomes intense cells and is assumed to be in a form similar to lymphoid tissue (including dendritic cells, T cells, B cells and NK cells, macrophages and plasma cell clusters). This process and a large number of immunopathological mechanisms including the formation of antigen-immunoglobulin complexes ultimately lead to the destruction of the integrity of the joint, which causes deformity, permanent loss of function and / or bone erosion at or near the joint. The disclosed compound, the combination of the disclosed compound or its composition can be used to treat, improve or prevent any single, several or all of these RA symptoms. Therefore, in the context of RA, when achieving the reduction or improvement of any symptom usually associated with RA, the compound is considered to provide a therapeutic benefit, regardless of whether the treatment results in a reduction in the amount of the combined treatment of the following RA and / or the circulating rheumatoid factor ("RF").

[0176] The American College of Rheumatology (ACR) has established criteria for defining improvement and clinical remission of RA. Once such parameters are achieved, ACR20 (ACR criteria for 20% clinical improvement) requires a 20% improvement in the counts of painful and swollen joints, as well as a 20% improvement in 3 of the following 5 parameters: patient global assessment, physician global assessment, patient pain assessment, degree of disability, and acute phase reactant levels. These criteria are improved by 50% and 70% in ACR50 and ACR70, respectively. Other criteria include Paulu's criteria and radiographic progression (e.g., Sharp score).

[0177] In some embodiments, a patient with RA obtains a therapeutic benefit when the patient exhibits an ACR of 20. In specific embodiments, an ACR improvement of ACR 50 or even ACR 70 may be achieved.

[0178] Cytokine release syndrome (CRS) is a potentially life-threatening condition that may be caused by a variety of factors, including severe viral infections (such as influenza), administration of antibodies for immunotherapy (such as cancer immunotherapy), and administration of non-protein-based cancer drugs (such as oxaliplatin and lenalidomide). Immunotherapy may involve high levels of immune activation that exceed naturally occurring immune activation levels, and CRS is a non-antigen-specific toxicity that may occur as a result. As immune-based therapies become more effective, the diagnosis rate of CRS is increasing. CRS has also been observed in cases of haploidentical donor stem cell transplantation and graft-versus-host disease. Shimabukuro-Vornhagen et al., Journal for ImmunoTherapy of Cancer 6:56 (2018). CRS is associated with elevated circulating levels of several cytokines including interleukin (IL)-6 and interferon gamma. Lee et al., Blood 124 (2): 188–195 (July 10, 2014; Electronic publication May 29, 2014).

[0179] CRS is often observed clinically when a large number of lymphocytes and / or myeloid cells are activated and release inflammatory cytokines. Cytokine release can be induced by chemotherapy or biological therapy, and / or can be associated with therapeutic antibody therapy (such as immunotherapy for cancer treatment). Exemplary immunotherapies that may cause CRS include, but are not limited to, therapies in which cells express recombinant receptors such as chimeric antigen receptors (CARs) and / or other transgenic receptors such as T cell receptors (TCRs). CRS induced by CAR T therapy usually occurs at the peak of CAR T cell expansion within a few days of T cell infusion. Giavridis et al., Nat Med. 24 (6): 731-738 (June 2018; electronic publication May 28, 2018). Examples of CAR T therapies that can induce CRS include sirolimus-acaptaline (with and tisagenlecleucel (marketed as Sale).

[0180] Highly elevated levels of interleukin 6 (IL-6) have been observed in CRS patients and in murine models of the disease, suggesting that IL-6 may play a role in the pathophysiology of CRS. Shimabukuro-Vornhagen, J Immunother Cancer 6(1), 56(2018). IL-6 can signal via two different modes. Classical IL-6 signaling involves the binding of IL-6 to the membrane-bound IL-6 receptor. However, the IL-6 receptor does not have an intracellular signaling domain. Instead, after soluble IL-6 binds to the membrane-bound IL-6 receptor, the IL-6 / IL-6 receptor complex binds to the membrane-bound gp130, which initiates signaling through its intracellular domain. In trans-signaling, IL-6 binds to a soluble form of the IL-6 receptor, which is typically cleaved from the cell surface by metalloproteinases. The resulting soluble IL-6 / IL-6 receptor complex binds to gp130 and can therefore also induce signaling in cell types that do not express the membrane-bound IL-6 receptor.

[0181] IL-6 contributes to many key symptoms of CRS. Through trans-signaling, IL-6 causes characteristic symptoms of severe CRS, i.e., vascular leakage, and activation of complement and coagulation cascades, thereby inducing disseminated intravascular coagulation (DIC). In addition, IL-6 may cause cardiomyopathy, which is often observed in CRS patients, by promoting myocardial dysfunction. In murine models, CRS appears within 2-3 days of CAR T cell infusion and may be fatal. Giavridis et al., Nat Med.24(6):731-738(2018). CRS symptoms may begin to appear within a few minutes or hours of the start of antibody treatment, and may include fever (may reach or exceed 40 ° C), nausea, fatigue, headache, tachycardia, hypotension, rash, shortness of breath and / or myalgia. However, in some cases, additional and potentially more serious complications may occur, including cardiac insufficiency, adult respiratory distress syndrome, neurotoxicity, renal and / or liver failure and / or disseminated intravascular coagulation.

[0182] The National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE v.5.0, published on November 27, 2017) includes a grading system for CRS.

[0183] Grade 1: Fever with or without systemic symptoms.

[0184] Grade 2: fluid-responsive hypotension; <40% O2-responsive hypoxia.

[0185] Grade 3: Hypotension controlled with one pressor; hypoxia requiring ≥ 40% O2.

[0186] Level 4: Life-threatening consequences; urgent intervention indicated.

[0187] Level 5: Death.

[0188] The subject may not exhibit signs or symptoms of CRS and / or may be at risk of developing CRS. In such embodiments, administration of the compound substantially prevents the onset of CRS, or prevents the onset of Grade 2 or higher CRS.

[0189] Alternatively, the subject exhibits at least one sign or symptom of CRS and may exhibit at least one sign or symptom of grade 1 CRS. Or the subject may exhibit at least one sign or symptom of grade 2 or higher CRS (such as grade 3 or higher CRS). The disclosed one or more compounds may be administered within 24 hours of the onset of signs or symptoms, and / or the administration of the one or more compounds may improve the signs or symptoms of CRS, such as reducing the grade of CRS from 4 to 3, 2 or 1, or from 3 to 2 or 1, or from 2 to 1, compared to the severity of the signs or symptoms before the administration of the one or more compounds. Alternatively, the CRS symptoms are substantially reduced to below grade 1, so that the subject no longer experiences symptoms associated with CRS. In some embodiments, the sign or symptom is a fever and may be a fever of 40°C or higher.

[0190] One or more compounds disclosed may be administered to a subject who has previously been administered a first therapy in which CRS is a known, suspected or potential side effect. Administration of the first therapy may begin greater than zero to 10 days before administration of the one or more compounds. Alternatively, one or more compounds may be administered to a subject who is about to or is being administered a first therapy in which CRS is a known, suspected and / or potential side effect. The first therapy may include cell therapy, including but not limited to chimeric antigen receptor (CAR) expression therapy and / or transgenic receptor therapy. It is also known that cell-free antibodies can cause this syndrome, especially those that activate T cells, including but not limited to CAMPATH 1-H, blinatumomab and / or rituximab.

[0191] A second therapeutic agent (e.g., a steroid, an anti-inflammatory agent, an immunosuppressant, or a combination thereof) may also be administered to treat or prevent CRS. The steroid may be a corticosteroid, such as, for example, dexamethasone or prednisone, or a combination thereof. The disclosed one or more compounds may be administered substantially simultaneously with the second therapeutic agent, or the one or more compounds and the second therapeutic agent may be administered sequentially in any order.

[0192] Acute respiratory distress syndrome (ARDS) is a syndrome characterized by severe shortness of breath, labored and unusually rapid breathing, hypotension, confusion, and extreme tiredness. 2O but PaO 2 / FiO 2 A ratio below 300 mmHg is diagnostic of this syndrome (Fan et al. JAMA. 319:698–71).

[0193] ARDS occurs when fluid builds up in the alveoli of the lungs. The fluid prevents the lungs from filling with enough air, which limits the amount of oxygen reaching the bloodstream, depriving organs of the oxygen they need to function. The intensity of ARDS symptoms can vary, depending on their cause and severity. Severe shortness of breath—the hallmark of ARDS—typically develops within hours to days after some respiratory viral infections, such as COVID-19 infection and influenza. Many people who develop ARDS do not survive, and the risk of death increases with age and severity of the disease. Among patients who survive ARDS, some recover fully, while others suffer lasting damage to their lungs. In some publications, ARDS may be referred to as acute lung injury (ALI).

[0194] Acute kidney injury (AKI), also known as acute renal injury (ARI) or acute renal failure (ARF), is a syndrome characterized by a sudden decline in renal function (including, for example, the ability to excrete waste from the patient's blood). AKI is characterized by a decline in glomerular filtration rate, urine output, or both. The loss of this filtering capacity leads to the retention of nitrogenous (urea and creatinine) and non-nitrogenous wastes that are usually excreted by the kidneys, a decrease in urine output, or both. Depending on the cause and effect relationship, AKI can be classified as prerenal, endogenous renal, or postrenal. Endogenous nephropathy can be further divided into glomerular, tubular, interstitial, and vascular abnormalities. AKI is accompanied by an inflammatory response that, if not inhibited, can lead to renal fibrosis and chronic renal failure. AKI usually exists over a period of hours or days and is potentially reversible. AKI can be characterized by an abrupt (i.e., within 14 days, 7 days, 72 hours, or 48 hours) decline in renal function identified by an absolute increase in serum creatinine greater than or equal to 0.3 mg / dl (≥26.4 μmol / l), a percentage increase in serum creatinine greater than or equal to 50% (1.5-fold relative to baseline), or a decrease in urine output (documented oliguria of less than 0.5 ml / kg per hour for at least 6 hours). Risk factors include, for example, subjects who are undergoing or have undergone major vascular surgery, coronary artery bypass grafting or other cardiac surgery; subjects with pre-existing congestive heart failure, preeclampsia, eclampsia, diabetes, hypertension, coronary artery disease, proteinuria, renal insufficiency, glomerular filtration below the normal range, cirrhosis, serum creatinine above the normal range, or sepsis; or subjects exposed to NSAIDs, cyclosporine, tacrolimus, aminoglycosides, foscarnet, ethylene glycol, heme, myoglobin, ifosfamide, heavy metals, methotrexate, radiopaque contrast agents, or streptozotocin. This list is not intended to be limiting.

[0195] Renal insufficiency includes, but is not limited to, kidney disorders, kidney disease, renal dysfunction, kidney cancer, other conditions where at least one kidney is missing or one or both kidneys do not function properly due to accident, surgical removal, or genetic disorder. Renal insufficiency can include acute kidney injury.

[0196] Thrombosis is a kind of coagulation disorder, and too many platelets can cause thrombosis.Thrombosis can refer to the formation of thrombus (blood clot) in blood vessels.The term covers but is not limited to arterial and venous thrombosis, including deep vein thrombosis, portal vein thrombosis, jugular vein thrombosis, renal vein thrombosis, stroke, myocardial infarction, Budd-Chiari syndrome (Budd-Chiari syndrome), Paget-Schroetter disease (Paget-Schroetter disease) and cerebral venous sinus thrombosis.In some embodiments, the patient is at a risk of thrombotic events relative to the general population (for example, measured by recognized risk factors).In some embodiments, the patient has one or more risk factors, and the risk factors make the patient have a high risk of thrombosis relative to the general population. Risk factors for thrombosis include, for example, classic cardiovascular risk factors: hyperlipidemia, smoking, diabetes, hypertension, and abdominal obesity; strong classic venous thromboembolism risk factors: trauma or fracture, major orthopedic surgery, and tumor surgery; moderate classic venous thromboembolism risk factors: non-oncological surgery, oral contraceptives and hormone replacement therapy, pregnancy and the postpartum period, hypercoagulable states, and previous venous thromboembolism; and weaker classic venous thromboembolism risk factors: age, bed rest (>3 days), long-term travel, and metabolic syndrome. Additional risk factors include hereditary, acquired, and mixed coagulation or metabolic risk factors for thrombosis, such as hereditary: antithrombin deficiency, protein C deficiency, protein S deficiency, Factor V Leiden, prothrombin G20210A; acquired: antiphospholipid syndrome; mixed: hyperhomocysteinemia, increased fibrinogen levels, increased factor VIII levels, increased factor IX levels. In some cases, the use of heparin may increase the risk of thrombosis, including, for example, heparin-induced thrombocytopenia (HIT). Diseases and disorders associated with thrombosis include, but are not limited to, acute venous thrombosis, pulmonary embolism, thrombosis during pregnancy, hemorrhagic skin necrosis, acute or chronic disseminated intravascular coagulation (DIC), sepsis-induced coagulopathy (SIC), thrombosis caused by surgery, prolonged bed rest, prolonged immobilization, venous thrombosis, fulminant meningococcal sepsis, acute thrombotic stroke, acute coronary artery occlusion, acute peripheral artery occlusion, massive pulmonary embolism, axillary vein thrombosis, massive iliofemoral vein thrombosis, arterial cannula occlusion, venous cannula occlusion, cardiomyopathy, hepatic veno-occlusive disease, hypotension, decreased cardiac output, decreased vascular resistance, pulmonary hypertension, decreased lung compliance, leukopenia, thrombocytopenia (e.g., immune thrombocytopenia) and immune thrombocytic purpura. In subjects at risk for thrombosis, the subject can be monitored using methods known to those of ordinary skill in the art to maintain hemostasis in patients at risk for thrombosis.Examples of methods for monitoring patients at risk for thrombosis include, but are not limited to, digital subtraction angiography, in vitro assays, or non-invasive methods. Examples of in vitro assays that can be used to identify and monitor subjects at risk for thrombosis and for treatment using the methods of the invention include, but are not limited to, functional assays and antibody detection assays.

[0197] A thrombotic event is any condition involving an arterial or venous obstruction or partial obstruction with thrombosis. Thrombotic events include, but are not limited to, thrombotic conditions such as myocardial infarction, unstable angina, stroke, pulmonary embolism, transient ischemic attack, deep vein thrombosis, reocclusion of a thrombus, and peripheral vascular thrombosis. Thrombotic events also include reocclusion of a thrombus after coronary intervention or thrombolytic therapy.

[0198] COVID-19 is the disease caused by infection with SARS-CoV-2 (formerly known as 2019-nCoV), which first appeared in Wuhan, China.

[0199] COVID-19-associated ARDS refers to ARDS caused by infection with SARS-CoV-2. People with COVID-19-associated ARDS may have been diagnosed with COVID-19, may have been exposed to another person with COVID19, or may be suspected of having COVID-19 based on their symptoms.

[0200] COVID-19-associated AKI refers to AKI caused by infection with SARS-CoV-2. Patients with COVID-19-associated AKI may have been diagnosed with COVID-19, may have been exposed to another person with COVID-19, or may be suspected of having COVID-19 based on their symptoms. In some cases, COVID-19-associated AKI includes AKI with symptoms such as those described in Batlle et al. J. AM. SOC. NEPHROL. 2020, 31(7): 1380-1383 and Gabarre et al. Intensive Care Med. 2020, 46(7): 1339-1348, the disclosures of which are incorporated herein by reference in their entirety.

[0201] COVID-19-associated thrombosis refers to thrombosis caused by SARS-CoV-2 infection. Patients with COVID-19-associated thrombosis may have been diagnosed with COVID-19, may have been exposed to another person with COVID-19, or may be suspected of having COVID-19 based on their symptoms. In some cases, COVID-19-associated thrombosis includes any of the symptoms described, for example, in Connors et al. Blood 2020, 135(23):2033-2040 and Bikdeli et al. J. Am. Coll. Cardiol. 2020, 75(23):2950-73, the disclosures of which are incorporated herein by reference in their entirety.

[0202] The term “COVID-19-related” refers to symptoms or indications that develop within 28 days of hospitalization due to signs of COVID-19 / COVID-19.

[0203] For COVID-19-related ARDS, successful treatment may include less shortness of breath, less difficulty breathing or rapid breathing, higher blood pressure, less confusion, and / or less fatigue. Treatment may be given prophylactically, that is, before ARDS develops. Prophylactic treatments may prevent ARDS and may be given to patients who have or are suspected of having COVID-19 but do not have severe symptoms of ARDS. For example, prophylactic treatments may be given to patients who have a cough but do not have other symptoms of ARDS.

[0204] For COVID-19 related AKI, successful treatment may include increased renal function. Renal function can be assessed by measuring serum creatinine levels, serum creatinine clearance, or blood urea nitrogen levels. In some cases, successful treatment includes a decline in metabolic acidosis, hyperkalemia, oliguria or anuria, azotemia, restoration of fluid balance, and improved effects on other organ systems. Treatment may be administered prophylactically, i.e., before the onset of AKI. Prophylactic treatment can prevent AKI and can be administered to patients who have been infected or suspected of being infected with COVID-19 but do not have severe symptoms of AKI. For example, prophylactic treatment may be administered to patients with one or more of the following: increased serum or urine creatinine, hematuria, hypoproteinemia, decreased antithrombin III levels, hypoalbuminemia, leukocyturia, or proteinuria without other AKI symptoms.

[0205] For COVID-19 related thrombosis, successful treatment can include improving the coagulation profile of the subject, or preventing, slowing down, delaying or preventing the subject from having a risky coagulation profile deterioration. Coagulation profile can be assessed by measuring one or more coagulation parameters, including one or more levels of D-dimer, factor II, factor V (e.g., Leiden Fifth Factor), factor VII, factor VIII, factor IX, factor XI, factor XII, factor XIII, F / fibrin degradation products, thrombin-antithrombin 111 complexes, fibrinogen, profibrinolysin, prothrombin and von Willebrand factor in the serum of the subject. Other coagulation parameters that can be measured for coagulation profile include, for example, prothrombin time, thromboplastin time, activated partial thromboplastin time (aPTT), antithrombin activity, platelet count, protein C level and protein S level. In addition, the C-reactive protein level of the patient can also be assessed before treatment, and if raised, this can be used as a further indicator of the increased risk of patient thrombosis.

[0206] Sepsis is a clinical syndrome of life-threatening organ dysfunction caused by a dysregulated immune response to infection. The more severe form of sepsis, "septic shock," is characterized by severely reduced tissue perfusion; acute failure of multiple organs, including the lungs, kidneys, and liver. Common causes in immunocompetent patients include many different species of Gram-positive and Gram-negative bacteria. Immunocompromised patients may have unusual bacterial or fungal species as causes. Signs include fever, hypotension, oliguria, and confusion. Diagnosis is primarily clinical in combination with culture results showing infection; early recognition and treatment are critical. Treatment is aggressive fluid resuscitation, antibiotics, surgical removal of infected or necrotic tissue and drainage of pus, and supportive care.

[0207] Influenza is a disease commonly referred to as "flu". Influenza is caused by a group of viruses that can be divided into four separate groups: influenza A, influenza B, influenza C and influenza D, distinguished by their nucleoprotein and matrix proteins. Influenza causes a viral respiratory infection, resulting in fever, rhinitis, cough, headache and malaise. Influenza A, B and C all infect humans, while there are no documented cases of influenza D infection in humans. On the other hand, influenza C does not cause the typical influenza illness seen in individuals infected with influenza A, B or C.

[0208] Influenza A strains are further classified based on two surface proteins, hemagglutinin (H) and neuraminidase (N). There are 18 different hemagglutinin subtypes and 11 different neuraminidase subtypes (H1 to H18 and N1 to N11, respectively). Although there are 198 different combinations of influenza A subtypes, only 131 subtypes have been detected in nature. The influenza A virus subtypes that are currently circulating routinely in the human population include: A(H1N1) and A(H3N2).

[0209] Cytokine release-related illness associated with influenza refers to any illness associated with influenza that results in high levels of cytokine release in the lungs and / or kidneys. Cytokine release-related illnesses include, but are not limited to, influenza-related ARDS, influenza-related AKI, influenza-related thrombosis, influenza-related sepsis, influenza-related septic shock, and the like.

[0210] Influenza-associated ARDS is ARDS caused by influenza infection. People with influenza-associated ARDS may have been diagnosed with influenza, may have been exposed to another person who has influenza, or may be suspected of having influenza based on their symptoms.

[0211] Influenza-associated AKI is AKI caused by influenza infection. Patients with influenza-associated AKI may have been diagnosed with influenza, may have been exposed to another person who has been infected with influenza, or may be suspected of being infected with influenza based on their symptoms. In some cases, influenza-associated AKI includes AKI with symptoms such as those described in Batlle et al. J. AM. SOC. NEPHROL. 2020, 31 (7): 1380-1383 and Gabarre et al. Intensive Care Med. 2020, 46 (7): 1339-1348, the disclosures of which are incorporated herein by reference in their entirety.

[0212] Influenza-associated thrombosis is thrombosis caused by influenza infection. Patients with influenza-associated thrombosis may have been diagnosed with influenza, may have been exposed to another person who has been infected with influenza, or may be suspected of being infected with influenza based on their symptoms. In some cases, influenza-associated thrombosis includes any of the symptoms described, for example, in Connors et al. Blood 2020, 135(23):2033-2040 and Bikdeli et al. J. Am. Coll. Cardiol. 2020, 75(23):2950-73, the disclosures of which are incorporated herein by reference in their entirety.

[0213] Influenza-associated sepsis is sepsis caused by an influenza infection. Patients with influenza-associated sepsis may have been diagnosed with influenza, may have been exposed to another person who has influenza, or may be suspected of having influenza based on their symptoms. In some cases, influenza-associated thrombosis includes symptoms such as those described in Florescu et al. Virulence. 2014 Jan 1; 5(1): 137–142. and Gu et al. Eur Respir Rev. 2020 Jul 21; 29(157): 200038, the disclosures of which are incorporated herein by reference in their entirety.

[0214] The term “influenza-associated” refers to symptoms or indications that develop within 28 days of hospitalization / signs of influenza infection.

[0215] For influenza-associated ARDS, successful treatment may include less shortness of breath, less difficulty breathing or rapid breathing, higher blood pressure, less confusion, and / or less fatigue. Treatment may be administered prophylactically, i.e., before the onset of ARDS. Prophylactic treatments can prevent ARDS and may be administered to patients who have been infected or suspected of being infected with the flu but do not have severe symptoms of ARDS. For example, prophylactic treatments may be administered to patients who have a cough but do not have other symptoms of ARDS.

[0216] For influenza-related AKI, successful treatment may include increased renal function. Renal function may be assessed by measuring serum creatinine levels, serum creatinine clearance or blood urea nitrogen levels. In some cases, successful treatment includes a decline in metabolic acidosis, hyperkalemia, oliguria or anuria, azotemia, the recovery of fluid balance and the improved effects on other organ systems. Treatment may be administered prophylactically, i.e., before the onset of AKI. Prophylactic treatment may prevent AKI and may be administered to patients who have been infected or suspected of being infected with influenza but do not have severe symptoms of AKI. For example, prophylactic treatment may be administered to patients with one or more of the following: serum or urine creatinine increases, hematuria, hypoproteinemia, antithrombin III levels decrease, hypoalbuminemia, leukocyturia or proteinuria without other AKI symptoms.

[0217] For influenza-related thrombosis, successful treatment can include improving the coagulation profile of experimenter, or preventing, slowing down, delaying or stopping experimenter from having the coagulation profile deterioration of risk.Coagulation profile can be assessed by measuring one or more coagulation parameters, including one or more levels of D-dimer, factor II, factor V (for example, Leiden Fifth Factor), factor VII, factor VIII, factor IX, factor XI, factor XII, factor XIII, F / fibrin degradation products, thrombin-antithrombin 111 complexes, fibrinogen, profibrinolysin, prothrombin and von Willebrand factor in the serum of experimenter.Other coagulation parameters that can be measured for coagulation profile include for example prothrombin time, thromboplastin time, activated partial thromboplastin time (aPTT), antithrombin activity, platelet count, protein C level and protein S level.In addition, before treatment, the C-reactive protein level of patient can also be assessed, and if raising, this can be used as the further index that patient's thrombosis risk increases.

[0218] For influenza-associated sepsis or septic shock, successful treatment may include a reduction in fever, a reduction in high or moderately high heartbeat (e.g., tachycardia), a reduction in sweating (i.e., diaphoresis), a reduction in confusion, and / or a reduction in fatigue and / or a reduction in shortness of breath, difficulty breathing, or rapid breathing. Treatment may be administered prophylactically, i.e., before the onset of sepsis or septic shock. Prophylactic treatments can prevent sepsis or septic shock and can be administered to patients who have been infected or suspected of being infected with influenza but do not have severe symptoms of sepsis or septic shock. For example, a prophylactic treatment may be administered to a patient who has a cough but does not have other symptoms of sepsis or septic shock.

[0219] In addition, the disclosed compounds, combinations of disclosed compounds, or compositions thereof can be used to treat sickle cell disease, particularly to reduce the immune response exhibited in the disease. In some embodiments, the subject may exhibit one or more of the following cancers: anemia, sickle cell crisis, vaso-occlusive crisis, splenic partition crisis, splenic partition crisis, acute chest syndrome, acute chest syndrome, aplastic crisis, hemolytic crisis, dactylitis, pneumonia, respiratory tract infection, bone marrow embolism, or atelectasis.

[0220] Sickle cell disease (SCD) is a group of usually inherited blood disorders. The most common type is called sickle cell anemia, which causes an abnormality in the oxygen-carrying protein hemoglobin found in red blood cells. In some cases, this results in a rigid sickle-like shape. Problems with sickle cell disease usually begin between 5 and 6 months of age, and a number of health problems may occur, such as painful episodes (called sickle cell crises), anemia, swelling of the hands and feet, bacterial infections, and strokes. As children age, long-term pain may develop.

[0221] Sickle cell disease occurs when a person inherits two abnormal copies (one from each parent) of the beta globin gene (HBB), which produces heme. The gene appears on chromosome 11. There are several subtypes, depending on the exact mutation in each heme gene. Attacks may be triggered by temperature changes, stress, dehydration, and high altitude.

[0222] Care for people with sickle cell disease can include preventing infection with vaccinations and antibiotics, plenty of fluid intake, folic acid supplements, and pain medications. Other measures may include blood transfusions and the drug hydroxycarbamide (hydroxyurea). A small percentage of people can be cured with a bone marrow cell transplant. People with sickle cell disease may experience the following symptoms:

[0223] Sickle cell crisis: The term "sickle cell crisis" or "sickling crisis" can be used to describe several independent acute episodes occurring in SCD subjects that result in anemia and crises, which can be of various types, including vaso-occlusive crisis, aplastic crisis, splenic partition crisis, hemolytic crisis, etc. Most sickle cell crisis episodes last five to seven days. In most cases, no precipitating cause is identified, although infection, dehydration, and acidosis (all of which favor sickling) may serve as triggering factors.

[0224] Vaso-occlusive crises: Vaso-occlusive crises are caused by sickle-shaped red blood cells that block capillaries and restrict blood flow to organs, resulting in ischemia, pain, necrosis, and often organ damage. The frequency, severity, and duration of these crises vary widely. Painful crises are treated with hydration, analgesia, and blood transfusions; pain management requires regular administration of opioids until the crisis resolves. For milder crises, a subset of subjects are managed with nonsteroidal anti-inflammatory drugs, such as diclofenac or naproxen. For more severe crises, most subjects require intra-subject administration of intravenous opioids; in these cases, a subject-controlled analgesia device is often used. Vaso-occlusive crises involving organs such as the penis or lungs are considered emergencies and are treated with red blood cell transfusions. Incentive spirometry, a technique that encourages deep breathing to minimize the development of atelectasis, is recommended.

[0225] Splenic partition crisis: Sickle cell disease often affects the spleen because sickle-shaped red blood cells narrow the blood vessels and reduce clearance of defective cells. Patients with sickle cell anemia often develop infarctions before the end of childhood. This splenic damage increases the risk of infection by encapsulated microorganisms; prophylactic antibiotics and vaccinations are recommended for those patients who lack a normal spleen function.

[0226] Splenic compartmental crises are acute, painful enlargement of the spleen caused by trapping of red blood cells within the spleen, which results in a dramatic drop in hemoglobin levels and may lead to hypovolemic shock. Compartmental crises are considered emergencies. Without treatment, the subject may die within 1–2 hours from circulatory collapse. Management is supportive, sometimes with transfusions. These crises are transient; they last 3–4 hours and may last up to a day.

[0227] Acute chest syndrome: Acute chest syndrome is defined by at least two of the following signs or symptoms: chest pain, fever, pulmonary infiltrates or focal abnormalities, respiratory symptoms, or hypoxemia. It is the second most common complication and accounts for approximately 25% of deaths in SCD subjects. Most cases present with a vaso-occlusive crisis and then develop acute chest syndrome. However, approximately 80% of people have a vaso-occlusive crisis during acute chest syndrome.

[0228] Aplastic Crisis: An aplastic crisis is an instance of acute worsening of a subject's baseline anemia, resulting in a pale appearance, increased heart rate, and fatigue. This crisis is usually triggered by the parvovirus B19, which directly affects the production of red blood cells by invading red blood cell precursors and multiplying and destroying them there. Parvovirus infection almost completely stops red blood cell production, lasting two to three days. In a normal individual, this is not significant, but the shortened lifespan of red blood cells in SCD subjects results in a sudden, life-threatening condition. During the illness, the reticulocyte count drops dramatically (causing reticulocytopenia), and the rapid turnover of red blood cells causes a drop in hemoglobin. This crisis takes 4 to 7 days to resolve. Most subjects can be managed supportively; some require blood transfusions.

[0229] Hemolytic Crisis: A hemolytic crisis is an acute, accelerated drop in hemoglobin levels. Red blood cells break down at a faster rate. This is especially common in people with coexisting G6PD deficiency. Another effect of hemolytic crisis in sickle cell disease is oxidative stress on red blood cells, white blood cells, and platelets. When not enough red blood cells are produced in the bone marrow, the oxygen that the body receives, processes, and transports is not balanced with the body's antioxidants. Oxygen activity in the cells is unbalanced, which results in more red blood cells being produced that are not properly oxygenated or formed. Oxidative stress may result in anemia due to the oxygen imbalance in the tissues. Management is supportive and sometimes transfusions.

[0230] In addition, one of the earliest clinical manifestations is dactylitis, which occurs as early as six months of age and may occur in children with sickle cell trait. The crisis may last up to one month. Given that both pneumonia and sickling in the lungs may produce symptoms of acute chest syndrome, subjects are treated for both disorders. It may be triggered by a painful crisis, respiratory infection, bone marrow embolism, or may be triggered by atelectasis, sedative administration, or surgery. Hematopoietic ulcers may also occur.

[0231] Additionally, the disclosed compounds, combinations of disclosed compounds, or compositions thereof can be used to treat lung injury.The lung injury can be chemical or radiation-induced lung injury.

[0232] In some embodiments, the subject may inhale or may be expected to be exposed to a lung irritant. In some embodiments, the subject may inhale or may be expected to inhale an asphyxiant. Pulmonary agents or asphyxiants are chemical agents intended to hinder the subject's ability to breathe. These compounds usually work by causing fluid accumulation in the lungs, which then leads to suffocation. Inhalation of these agents can cause burning throat, coughing, vomiting, headache, chest pain, chest tightness, and respiratory and circulatory failure. Examples of such agents include: chlorine, chloropicrin (PS), diphosgene (DP), phosgene (CG), sulfur pentafluoride, perfluoroisobutylene, acrolein, and diphenyl cyanoarsine. Phosgene-induced acute lung injury (P-ALI) is usually associated with short-term phosgene inhalation. Long-term exposure may lead to chronic hypoventilation, refractory pulmonary edema, and other related lung injuries, ultimately leading to ARDS. Chemical pneumonia is lung inflammation or dyspnea caused by inhaling chemical fumes or breathing in certain chemicals and suffocating.

[0233] Additionally, the disclosed compounds, combinations of disclosed compounds, or compositions thereof can be used to treat or prevent acute inhalation injury (AII) and electronic cigarette or hookah product use-associated lung injury (EVALI).

[0234] In other embodiments, the subject has been exposed to or is expected to be exposed to ionizing radiation. In these embodiments, the subject may have or may be expected to develop radiation-induced lung injury (RILI). In some embodiments, the subject may suffer from radiation pneumonitis or radiation pulmonary fibrosis. In these embodiments, the subject may have received or is receiving chest radiation therapy, may have inhaled radioactive agents or may have been directly exposed to ionizing radiation. For example, the subject may have inhaled radioactive agents, or may have been directly exposed to ionizing radiation due to, for example, nuclear weapons or nuclear power plant leaks.

[0235] The disclosed compounds, combinations of disclosed compounds, or compositions thereof can be used to treat or prevent hemorrhagic fevers or symptoms thereof, including Ebola virus disease, Alkhurma hemorrhagic fever, Chapare hemorrhagic fever, Crimean-Congo hemorrhagic fever, Hantavirus pulmonary syndrome (HPS), hemorrhagic fever with renal syndrome (HFRS), Casanore Forest disease (KFD), Lassa fever, Lujo hemorrhagic fever, Marburg hemorrhagic fever, Omsk hemorrhagic fever, Rift Valley fever, yellow fever, or dengue fever, such as severe dengue (dengue hemorrhagic fever).

[0236] B. Formulation and Administration

[0237] The pharmaceutical composition comprising one or more active compounds of the present disclosure can be manufactured by any suitable method, such as mixing, dissolving, granulating, making sugar-coated tablets, grinding, emulsifying, encapsulating, embedding or lyophilizing processes. The composition can be formulated using one or more physiologically acceptable excipients, diluents, carriers, adjuvants or auxiliary agents to provide a pharmaceutically usable preparation.

[0238] One or more active compounds can be formulated in the form of a pharmaceutical composition itself, or in the form of a hydrate, solvate, N-oxide or a pharmaceutically acceptable salt. Generally, such salts are more soluble in aqueous solution than the corresponding free acids and bases, but salts with lower solubility than the corresponding free acids and bases can also be formed.

[0239] Pharmaceutical compositions containing one or more of the disclosed compounds can be in a form suitable for nearly any mode of administration including, for example, topical, ocular, oral, buccal, systemic, intranasal, injectable (such as intravenous or intraperitoneal), transdermal, rectal, vaginal, etc., or in a form suitable for administration by inhalation or insufflation.

[0240] For topical administration, one or more active compounds (or their hydrates, solvates, N-oxides or pharmaceutically acceptable salts) may be formulated into solutions, gels, ointments, creams, suspensions, etc. as are well known in the art.

[0241] Systemic formulations include those designed for administration by injection (eg, subcutaneous, intravenous, intramuscular, intrathecal, or intraperitoneal injection), as well as those designed for transdermal, transmucosal, oral, or pulmonary administration.

[0242] Useful injectable formulations include sterile suspensions, solutions or emulsions of one or more active compounds in aqueous or oily vehicles. The composition may also contain preparatants, such as suspending agents, stabilizers and / or dispersants. The formulations for injection may be present in unit dosage form, for example, in ampoules or in multidose containers, and may contain added preservatives.

[0243] Alternatively, injectable preparations may be provided in powder form for reconstitution with a suitable vehicle, including but not limited to sterile, pyrogen-free water, buffer, dextrose solution, etc., prior to use. To this end, one or more active compounds may be dried by any technique known in the art, such as lyophilization, and reconstituted prior to use.

[0244] For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are known in the art.

[0245] For oral administration, the pharmaceutical composition can take the form of, for example, lozenges, tablets or capsules prepared by conventional means with pharmaceutically acceptable excipients such as binders (e.g., pregelatinized corn starch, polyvinyl pyrrolidone or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc or silicon dioxide); disintegrants (e.g., potato starch or sodium starch glycolate); and / or wetting agents (e.g., sodium lauryl sulfate). Tablets can be coated by methods well known in the art, for example, with sugar, film or enteric coatings.

[0246] Liquid preparations for oral administration may take the form of, for example, elixirs, solutions, syrups or suspensions, or they may be presented as a dry product for constitution with water or other suitable vehicles before use. Such liquid preparations may be prepared by conventional means with pharmaceutically acceptable excipients such as suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifiers (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethanol, cremophore TM or fractionated vegetable oil); and a preservative (for example, methyl or propyl p-hydroxybenzoate or sorbic acid). The preparation may also contain buffer salts, preservatives, flavoring agents, coloring agents and sweeteners as appropriate.

[0247] Preparations for oral administration may be suitably formulated to give controlled release of the active compound, as is well known.

[0248] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0249] For rectal and vaginal routes of administration, the active compound or compounds may be formulated as solutions (for retention enemas), suppositories, or ointments containing conventional suppository bases such as cocoa butter or other glycerides.

[0250] For intranasal administration or administration by inhalation or insufflation, one or more active compounds, hydrates, solvates, N-oxides or pharmaceutically acceptable salts may be conveniently delivered as an aerosol from a pressurized pack or nebulizer using a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, fluorocarbons, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules or cartridges (for example, those comprising gelatin) for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base (such as lactose or starch).

[0251] The pharmaceutical composition may be in the form of a sterile injectable aqueous or oily suspension. The suspension may be prepared according to known techniques using those suitable dispersants or wetting agents and suspending agents already mentioned above. Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic, parenterally acceptable diluents or solvents. Among acceptable vehicles and solvents, water, Ringer's solution, and isotonic sodium chloride solution may be used.

[0252] According to the present disclosure, the disclosed one or more compounds, their solvates, N-oxides, pharmaceutically acceptable salts, or one or more prodrug forms may also be delivered by any of a variety of inhalation devices and methods known in the art, including, for example: U.S. Pat. No. 6,241,969; U.S. Pat. No. 6,060,069; U.S. Pat. No. 6,238,647; U.S. Pat. No. 6,335,316; U.S. Pat. No. 5,364,838; U.S. Pat. No. 5,672,581; WO 96 / 32149; WO 95 / 24183; U.S. Pat. No. 5,654,007; U.S. Pat. No. 5,404,871; U.S. Pat. No. 5,672,581; U.S. Pat. No. 5,743,250; U.S. Pat. No. 5,419,315; U.S. Pat. No. 5,558,085; WO 98 / 33480; U.S. Patent No. 5,364,833; U.S. Patent No. 5,320,094; U.S. Patent No. 5,780,014; U.S. Patent No. 5,658,878; 5,518,998; 5,506,203; U.S. Patent No. 5,661,130; U.S. Patent No. 5,655,523; U.S. Patent No. 5,645,051; U.S. Patent No. 5,622,166; U.S. Patent No. 5,577,497; U.S. Patent No. 5,492,112; U.S. Patent No. 5,327,883; U.S. Patent No. 5,277,195; U.S. Publication No. 20010041190; U.S. Publication No. 20020006901; and U.S. Publication No. 20020034477.

[0253] Devices that can be used to administer one or more forms of the active compound include those well known in the art such as metered dose inhalers, liquid nebulizers, dry powder inhalers, sprayers, thermal vaporizers, etc. Other suitable technologies for administering specific 2,4-pyrimidinediamine compounds include electrofluidic nebulizers.

[0254] In addition, the inhalation device is preferably practical in the sense of being easy to use, small enough to be portable, capable of providing multiple doses, and durable. Some specific examples of commercially available inhalation devices are Turbohaler (Astra, Wilmington, DE), Rotahaler (Glaxo, Research Triangle Park, NC), Diskus (Glaxo, Research Triangle Park, NC), Ultravent nebulizer (Mallinckrodt), Acorn II nebulizer (Marquest Medical Products, Totowa, NJ), Ventolin metered dose inhaler (Glaxo, Research Triangle Park, NC), etc. In one embodiment, the disclosed one or more compounds, their solvates, N-oxides, pharmaceutically acceptable salts, or one or more prodrugs can be delivered by a dry powder inhaler or a nebulizer.

[0255] As will be appreciated by those of ordinary skill in the art, the amount of the preparation of the form of one or more disclosed compounds, its solvates, N-oxides, pharmaceutically acceptable salts or one or more prodrugs, the preparation delivered and the application duration of a single dose depend on the type and other factors of the inhalation device used.For some aerosol delivery systems, such as nebulizers, the time length of the frequency of application and the system being activated will depend mainly on the concentration of one or more disclosed compounds in the aerosol.For example, a shorter application time can be used when the disclosed one or more compounds are at a higher concentration in the nebulizer solution.In some embodiments, a device such as a metered dose inhaler can produce higher aerosol concentrations, and a shorter time can be operated to deliver the active compound of the desired amount.Devices such as dry powder inhalers deliver active agents until the agent of a given load is discharged from the device.In this type of inhaler, the amount of one or more disclosed compounds, its solvates, N-oxides, pharmaceutically acceptable salts or one or more prodrugs in a given amount of powder determines the dosage delivered in a single application.Select the preparation of one or more disclosed compounds to produce a desired particle size in a selected inhalation device.

[0256] Formulations of the disclosed compounds for administration from a dry powder inhaler can generally include a finely divided dry powder containing one or more of the disclosed compounds, but the powder can also include a bulking agent, a buffer, a carrier, an excipient, another additive, etc. Additives can be included in dry powder formulations, for example, to dilute the powder as needed for delivery from a particular powder inhaler, to facilitate processing of the formulation, to provide favorable powder properties to the formulation, to facilitate dispersion of the powder from the inhalation device, to stabilize the formulation (e.g., antioxidants or buffers), to provide flavor to the formulation, etc. Typical additives include monosaccharides, disaccharides, and polysaccharides; sugar alcohols and other polyols, such as, for example, lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch, or combinations thereof; surfactants, such as sorbitol, diphosphatidylcholine, or lecithin; etc.

[0257] The disclosed method can be carried out with a pharmaceutical composition comprising one or more disclosed compounds suitable for administration by inhalation. For example, dry powder formulations can be prepared in a variety of ways using conventional techniques, such as those described in any of the above-mentioned publications and explicitly incorporated herein by reference, and, for example, U.S. Patent No. 5,700,904 of Baker et al., the disclosure of which is explicitly incorporated herein by reference. Particles within a size range suitable for maximum deposition in the lower respiratory tract can be prepared by micronization, grinding, etc. And liquid preparations can be prepared by dissolving the compound in a suitable solvent (such as water, at a suitable pH, including a buffer or other excipients).

[0258] A specific example of an aqueous suspension formulation suitable for intranasal administration using a commercially available intranasal spray device comprises the following ingredients: active compound (0.5 20 mg / ml); benzalkonium chloride (0.1 0.2 mg / mL); polysorbate 80 ( 80; 0.5 5mg / ml); sodium carboxymethylcellulose or microcrystalline cellulose (1 15mg / ml); phenylethanol (1 4mg / ml); and glucose (20 50mg / ml). The pH of the final suspension can be adjusted to a range of about pH 5 to pH 7, with a typical pH being about pH 5.5.

[0259] Another specific example of an aqueous suspension suitable for administration of a compound via inhalation contains 20 mg / mL of one or more disclosed compounds, 1% (v / v) polysorbate 80 ( 80), 50 mM citric acid and / or 0.9% sodium chloride.

[0260] For ocular administration, one or more active compounds can be formulated into solutions, emulsions, suspensions, etc. suitable for administration to the eye. Various vehicles suitable for administering compounds to the eye are known in the art. Specific non-limiting examples are described in the following: U.S. Patent Nos. 6,261,547; 6,197,934; 6,056,950; 5,800,807; 5,776,445; 5,698,219; 5,521,222; 5,403,841; 5,077,033; 4,882,150; and 4,738,851, which are incorporated herein by reference.

[0261] For extended delivery, one or more active compounds can be formulated into a reservoir preparation for use by implantation or intramuscular injection. Active component can be formulated with a suitable polymer or hydrophobic material (for example, as an emulsion in an acceptable oil) or an ion exchange resin, or be formulated into a slightly soluble derivative, for example, be formulated into a slightly soluble salt. Alternatively, the transdermal delivery system of an adhesive disc or a patch manufactured to slowly release one or more active compounds for percutaneous absorption can be used. For this reason, a penetration enhancer can be used to promote the transdermal penetration of one or more active compounds. Suitable transdermal patches are described, for example, in U.S. Pat. Nos. 5,407,713; 5,352,456; 5,332,213; 5,336,168; 5,290,561; 5,254,346; 5,164,189; 5,163,899; 5,088,977; 5,087,240; 5,008,110; and 4,921,475, which are incorporated herein by reference.

[0262] Alternatively, other drug delivery systems may be used. Liposomes and emulsions are well-known examples of delivery vehicles that can be used to deliver one or more active compounds. Certain organic solvents (such as dimethyl sulfoxide (DMSO)) may also be used, although typically at the expense of greater toxicity.

[0263] If desired, the pharmaceutical composition may be presented in a package or dispenser device which may include one or more unit dosage forms containing one or more active compounds. The package may, for example, include metal or plastic foil, such as a blister pack. The package or dispenser device may be accompanied by instructions for use.

[0264] C. Dosage

[0265] The disclosed compounds or combinations of disclosed compounds will generally be used in an amount effective to achieve the desired result, such as an amount effective to treat or improve a particular illness. The disclosed one or more compounds or pharmaceutical compositions thereof can be administered therapeutically to achieve a therapeutic benefit, or administered preventively to achieve a preventive benefit. Therapeutic benefit means the eradication or improvement of the underlying condition being treated and / or the eradication or improvement of one or more symptoms associated with the underlying condition, such that although the patient may still be afflicted by the underlying condition, the patient reports improvement in terms of sensation or illness. For example, administering a compound to a patient suffering from allergies provides a therapeutic benefit not only when the underlying allergic reaction is eradicated or improved, but also when the patient reports a reduction in the severity or duration of symptoms associated with the allergy after exposure to an allergen. As another example, the therapeutic benefit in the context of asthma includes an improvement in breathing after the onset of an asthma attack or a reduction in the frequency or severity of an asthma attack. Whether or not improvement is achieved, the therapeutic benefit also includes a stop or slowing of the progression of the disease.

[0266] As known by those of ordinary skill in the art, the preferred dosage of the disclosed compound may depend on various factors, including the age, weight, overall health and severity of the illness of the patient or subject being treated. When administered by inhalation, the dosage may also need to be adjusted to accommodate the gender of the individual and / or the lung capacity of the individual. The dosage may also be adjusted to accommodate individuals with more than one illness or individuals with other illnesses (e.g., emphysema, bronchitis, pneumonia and respiratory tract infections) that affect lung capacity and normal breathing ability. The dosage and frequency of administration of the disclosed one or more compounds or compositions thereof will also depend on whether the disclosed one or more compounds are formulated for the treatment of acute episodes of illness or for the preventive treatment of illness. One of ordinary skill in the art will be able to determine the optimal dosage for a particular individual.

[0267] For prophylactic administration, the disclosed compounds, combinations of disclosed compounds, or compositions thereof may be administered to a patient or subject at risk of developing one of the previously described disorders. For example, if it is not known whether the patient or subject is allergic to a particular drug, the disclosed compounds, combinations of disclosed compounds, or compositions thereof may be administered before the administration of the drug to avoid or improve allergic reactions to the drug. Alternatively, prophylactic administration may be used to avoid or improve the onset of symptoms in patients diagnosed with underlying conditions. For example, one or more disclosed compounds, or compositions thereof, may be administered to allergy victims prior to expected exposure to allergens. The disclosed compounds, combinations of disclosed compounds, or compositions thereof may also be prophylactically administered to healthy individuals to prevent the onset of conditions, the healthy individuals being repeatedly exposed to agents known to cause one of the above-mentioned diseases. For example, the disclosed compounds, combinations of disclosed compounds, or compositions thereof may be administered to healthy individuals repeatedly exposed to allergens known to cause allergies (such as latex) in an attempt to prevent individuals from developing allergies. Alternatively, the disclosed compounds, combinations of disclosed compounds, or compositions thereof may be administered to patients with asthma prior to participating in activities that trigger asthma attacks to reduce the severity of asthma attacks or to avoid asthma attacks altogether.

[0268] Effective doses can be estimated initially from in vitro assays. For example, an initial dose for a subject can be formulated to achieve an IC value equal to or greater than that of a particular compound as measured in an in vitro assay. 50 or EC 50 The circulating blood or serum concentration of the active compound is calculated to achieve such circulating blood or serum concentrations. Doses can be calculated to achieve such circulating blood or serum concentrations, taking into account the bioavailability of a particular compound. Fingl and Woodbury, "General Principles," In: Goodman and Gilman's The Pharmaceutical Basis of Therapeutics, Chapter 1, pp. 1-46, Pergamon Press, and references cited therein, provide additional guidance on effective doses.

[0269] In some embodiments, the disclosed compounds have an EC with respect to a kinase protein, such as an IRAK protein, from greater than 0 to 20 μM, such as greater than 0 to 10 μM, greater than 0 to 5 μM, greater than 0 to 1 μM, greater than 0 to 0.5 μM, greater than 0 to 0.1 μM, or greater than 0 to 0.05 μM. 50 .

[0270] Initial doses can also be estimated from in vivo data, such as animal models. Animal models that can be used to test the efficacy of compounds in treating or preventing the various diseases described above are well known in the art. Suitable animal models of hypersensitivity or allergic reactions are described in: Foster, (1995) Allergy 50 (Suppl. 21): 6-9, discussion 34-38; and Tumas et al., (2001), J. Allergy Clin. Immunol. 107 (6): 1025-1033. Suitable animal models of allergic rhinitis are described in Szelenyi et al., (2000), Arzneimittelforschung 50 (11): 1037-42; Kawaguchi et al., (1994), Clin. Exp. Allergy 24 (3): 238-244 and Sugimoto et al., (2000), Immunopharmacology 48 (1): 1-7. One of ordinary skill in the art can adapt such information to determine appropriate dosages for human administration.

[0271] The dosage of the disclosed compound will generally be greater than 0mg / kg / day (such as 0.0001mg / kg / day or 0.001mg / kg / day or 0.01mg / kg / day) up to at least about 100mg / kg / day in the range. More generally, dosage (or effective amount) can be in the range of about 0.0025mg / kg to about 1mg / kg administered at least once a day, such as 0.01mg / kg to about 0.5mg / kg or about 0.05mg / kg to about 0.15mg / kg. Total daily dose is generally in the range of about 0.1mg / kg to about 5mg / kg or to about 20mg / kg per day, such as 0.5mg / kg to about 10mg / kg per day or about 0.7mg / kg to about 2.5mg / kg / day per day. In addition to other factors, dosage can be higher or lower, depending on the activity of the disclosed compound, its bioavailability, mode of administration, and various factors discussed above.

[0272] Doses and dosage intervals can be adjusted for the individual to provide plasma levels of the disclosed compounds sufficient to maintain a therapeutic or preventive effect. For example, the compound can be administered once a day, multiple times a day, once a week, multiple times a week (e.g., every other day), once a month, multiple times a month, or once a year, depending on, among other things, the mode of administration, the specific indication being treated, and the judgment of the prescribing physician. One of ordinary skill in the art will be able to optimize the effective topical dose without undue experimentation.

[0273] Compositions comprising one or more disclosed compounds typically contain greater than 0% to up to 99% of the disclosed compound or compounds and / or other therapeutic agents (by total weight percentage). More generally, compositions comprising one or more disclosed compounds contain about 1 to about 20 total weight percentages of disclosed compounds and other therapeutic agents, and about 80 to about 99 weight percentages of pharmaceutically acceptable excipients. Typical daily administration can be within the range of 100–300 mg / day, such as 100, 150, 200, 250 or 300 mg / day. Administration can be once or twice or more per day, for example, 100 or 150 mg BID. Therefore, pharmaceutical dosage forms comprising compounds disclosed herein can contain 50–300 mg of disclosed compounds, such as 50, 100, 150, 200, 250, 300 mg of disclosed compounds.

[0274] Preferably, the disclosed compounds, combinations of disclosed compounds, or compositions thereof will provide therapeutic or preventive benefits without causing substantial toxicity. The toxicity of the disclosed compounds can be determined using standard pharmaceutical procedures. The dose ratio between toxicity and therapeutic (or preventive) effects is the therapeutic index. Disclosed compounds that exhibit high therapeutic indexes are preferred.

[0275] VII. Examples

[0276] Example 1

[0277] 3-(Dimethylamino)-1-(pyrimidin-2-yl)prop-2-en-1-one (D-2)

[0278]

[0279] A solution of 1-(pyrimidin-2-yl)ethane-1-one (D-1, 5 g, 40.95 mmol) in 1,1-dimethoxy-N,N-dimethylmethanamine (11 mL, 81.9 mmol) was stirred at 90 °C overnight. The reaction went to completion as monitored by LCMS. The volatiles were removed in vacuo and the crude product was used in the next reaction without further purification: 1 H NMR (400MHz, chloroform-d) δ8.89(d,J=4.9Hz,2H),8.00(d,J=10.3Hz,1H),7.35(dd,J=4 .9,4.9Hz,1H),6.39(d,J=10.3Hz,1H),3.20(s,3H),3.00(s,3H); LRMS(M+H)m / z 178.2.

[0280] Example 2

[0281] 2-(1H-Pyrazol-3-yl)pyrimidine (D-3)

[0282]

[0283] A hydrazine monohydrate solution (64%-65% aqueous solution, 2.8 mL, about 0.9 equiv) was added to a solution of 3-(dimethylamino)-1-(pyrimidin-2-yl)prop-2-en-1-one (D-2, 41 mmol) in EtOH (15 mL) and cooled in an ice bath. After reflux for 3 hours, the reaction went to completion as monitored by LCMS. The volatiles were removed in vacuo and the crude product was used in the next reaction without further purification; 1 H NMR(400MHz,chloroform-d)δ11.20(br s,1H),8.79(d,J=4.9Hz,2H),7.71(d,J=2.0Hz,1H),7.22(dd,J=4.9,4.9Hz,1H),7.08(d,J=2.0Hz,1H); LRMS(M+H)m / z147.1.

[0284] Example 3

[0285] 2-(4-Nitro-1H-pyrazol-3-yl)pyrimidine (D-4)

[0286]

[0287] 90% fuming nitric acid (3.8 mL, 81.9 mmol) was added dropwise to a concentrated H2O solution of 2-(1H-pyrazol-3-yl)pyrimidine (D-3, about 41 mmol). 2 SO 4 The mixture was stirred at 70 °C overnight. The reaction was complete as monitored by LCMS. After cooling to room temperature, the mixture was carefully poured into ice water. The pH was adjusted to 8 with aqueous NaOH solution, and the product was extracted with EtOAc (100 mL × 5). The combined organic layers were dried (Na 2 O 4 3 O 4 4 4 4 2 4 ... 2 SO 4 ), filtered, and the solvent was removed in vacuo. After silica gel chromatography, compound 2-(4-nitro-1H-pyrazol-3-yl)pyrimidine (D-4) was obtained as an off-white solid: 5.6 g (71.5% yield in 3 steps); 1 HNMR (400MHz, chloroform-d) δ 11.26 (br s, 1H), 8.90 (d, J = 4.9Hz, 2H), 8.28 (s, 1H), 7.40 (dd, J = 4.9, 4.9Hz, 1H); LRMS (M+H) m / z 192.1.

[0288] Alternative Procedure:

[0289] 2-(1H-pyrazol-3-yl)pyrimidine (D-3, 32.6 g, 223 mmol) was added to concentrated H 2 SO 4 Fuming nitric acid (100 mL) was added dropwise to the (200 mL) suspension, and the reaction mixture was stirred at room temperature overnight. The mixture was carefully poured onto ice, and a 27 N NaOH aqueous solution was added to adjust the pH to 6. The solid was collected by filtration, washed with water, and dried in vacuo. Compound 2- (4- nitro -1H- pyrazole -3- bases) pyrimidine was obtained as an off-white solid: 29.2 g (69% yield).

[0290] Example 4

[0291] 8-Ethoxy-1,4-dioxaspiro[4.5]decane (B-2)

[0292]

[0293] A solution of 1,4-dioxaspiro[4.5]decanol B-1 (158 g, 1.0 mol, 1.0 eq.) in tetrahydrofuran (500 mL) was added to a suspension of hexane-washed sodium hydride (48 g 60% suspension in mineral oil, 1.2 mol, 1.2 eq.) in tetrahydrofuran (500 mL) at 0 °C. The reactants were stirred at 0 °C for 15 minutes and at room temperature for 4 hours, then cooled to 0 °C and iodoethane (104.5 mL, 1.3 mol, 1.3 eq.) was added. The reactants were stirred at 0 °C for 10 minutes and at room temperature for 14 hours. The mixture was stirred for 14 hours at 37 °C. The mixture was stirred for 2 hours at 37 °C. The mixture was stirred for 2 hours at 37 °C. The mixture was stirred for 2 hours at 37 °C. The mixture was stirred for 2 hours at 37 °C. The mixture was stirred for 3 hours at 37 °C. The mixture was stirred for 2 hours at 37 °C. The mixture was stirred for 2 hours at 37 °C. The mixture was stirred for 2 hours at 37 °C. 4 Cl (about 100mL) was used to quench the reaction. The reaction was concentrated to remove tetrahydrofuran, and the concentrate was distributed between EtOAc (800mL) and water (600mL). The aqueous phase was extracted with EtOAc (300mL). The combined organic matter was washed with NaHCO 3 (500 mL) and brine (500 mL), dried (Na 2 SO 4 ), and concentrated under reduced pressure to give the title compound as an orange oil (177 g, 95%); 1 H nmr(400MHz,CD 3 Cl)δ3.90,3.89(4H,2dAB system,J 2.5Hz,OCH 2 CH 2 O), 3.44(2H,q,J 7.0Hz,OC H 2 CH 3),3.35(1H,tt,J 7.5,3.0Hz,H-8),1.82-1.74(4H,m,4H's H-6,H-7,H-9,H-10),1.70-1.63(2H,m,2H's H-6,H-7,H-9,H-10),1.54-1.47(2H,m,2H's H-6,H-7,H-9,H-10),1.15(3H,t,J 7.0Hz,OCH 2 C H 3 ).

[0294] Example 5

[0295] 4-Ethoxycyclohexane-1-one (B-3)

[0296]

[0297] Hydrochloric acid (450mL of 3M solution) was added to a solution of dioxane B-2 (177g, 952mmol) in tetrahydrofuran (450mL). The reaction was stirred vigorously at 60°C for 20 hours. NMR analysis showed 75% completion. Hydrochloric acid (100mL of 4M solution) was added again, and the reaction was stirred at 60°C for another 24 hours, then cooled and concentrated to remove tetrahydrofuran. The organic matter was extracted with EtOAc (3x 400mL). The combined organic matter was washed with brine (400mL), dried (Na2O3) and purified by column chromatography. 2 SO 4 ), and concentrated under reduced pressure to give the title compound (142.2 g, theoretical yield 135.1 g) as an orange oil; 1 H nmr(400MHz,CD 3 Cl)δ3.71(1H,m,cyclohexane H-4),3.55(2H,q,J 7.0Hz,OC H 2 CH 3 ),2.58(2H,ddd,J15.5,10.5,6.0Hz,2H cyclohexane H-2,H-6),2.25(2H,m,2H cyclohexane H-2,H-6),2.10-2.02(2H,m,2H cyclohexane H-3,H-5),1.98-1.90(2H,m,2H cyclohexane H-3,H-5),1.24(3H,t,J 7.0Hz,OCH 2 C H 3 ).

[0298] Example 6

[0299] (1s,4s)-4-ethoxycyclohexane-1-ol (B-4)

[0300]

[0301] A solution of ketone B-3 (146.2 g, 1.0 eq.) in tetrahydrofuran (300 mL) was cooled to -78 °C. Lithium aluminum hydride (400 mL of a 1 M solution in tetrahydrofuran, 400 mmol, 0.4 eq.) was added dropwise over 90 minutes. The reaction was stirred at -78 °C for another 30 minutes, and the reaction was quenched by initially adding NaOH (1 M, 300 mL) dropwise at -78 °C, then removed from the cold bath. A gel was produced. The reaction was diluted and stirred with EtOAc (400 mL), then decanted to remove the gel. The decanted organics were washed with Rochelle salt (400 mL), and the aqueous phase was stripped with EtOAc (200 mL). The combined organics were washed with Rochelle salt (300 mL) and brine (400 mL), dried (Na 2 SO 4 ), and concentrated under reduced pressure to obtain the title compound (131.1 g) as an orange oil without further purification; 1 H nmr(400MHz,CD 3 Cl) δ 3.74 (1H major product, m, cyclohexane H-1), 3.68 (1H minor product, m, cyclohexane H-1), 3.48 (2H minor product, J 7.0 Hz, OC H 2 CH 3 ), 3.47 (2H major product, J 7.0 Hz, OC H 2 CH 3 ),3.37(1H major product,tt,J 6.5,3.0Hz,H-4),3.25(1H minor product,m,H-4),1.99(1H,m,1H's H-2,H-3,H-5,H-6),1.81(2H,m,2H's H-2,H-3,H-5,H-6),1.73-1.51(5H,m,5H's H-2,H-3,H-5,H-6),1.19(3H major product,t,J 7.0Hz,OCH 2 C H 3 ), 1.19 (3H minor product, J 7.0 Hz, OCH 2 C H 3 ).

[0302] Example 7

[0303] 4-Nitrobenzenesulfonic acid (1r,4r)-4-ethoxycyclohexyl ester (B-5)

[0304]

[0305] To a solution of 4-ethoxycyclohexane-1-ol B-4 (112.9 g, 782.9 mmol, 1.0 eq. as 1s, 4s and 1r, 4s diastereomers in an approximate 2:1 ratio) in dichloromethane (800 mL) was added 1,4-diazabicyclo[2.2.2]octane (105.4 g, 939.5 mmol, 1.2 eq.) at 0°C. 4-Nitrobenzenesulfonyl chloride (190.8 g, 861.2 mmol, 1.1 eq.) was added portionwise at 0°C over 1 hour and the reaction was allowed to warm to room temperature over 16 hours. The reaction was heated with CH 2 Cl 2 (400 mL) and diluted with NaHCO 3 (1 L), water (1 L) and brine (1 L). The organics were dried (Na 2 SO 4 ) and concentrated under reduced pressure. The resulting solid was triturated from ethyl acetate and hexanes to yield 160.8 g of 4-ethoxycyclohexyl 4-nitrobenzenesulfonate (as a mixture of 1r, 4r and 1s, 4s) as an off-white solid; 1 H nmr(400MHz,CD 3 Cl)δ8.39(2H,d,J 9.0Hz,2H C 6 H 4 NO 2 ),8.11(2H,d,J 9.0Hz,2H C 6 H 4 NO 2 ),4.75(1H major product,tt,J 7.5,3.5Hz,H-1),4.70(1H minor product,m,H-1),3.45(2H minor product,J 7.0Hz,OC H 2 CH 3 ),3.44(2H main product,J7.0Hz,OC H 2 CH 3 ),3.34(1H,tt,J 6.5,3.0Hz,H-4),1.99-1.88(3H,m,3H's H-2,H-3,H-5,H-6),1.80-1.71(1H,m,1H's H-2,H-3,H-5,H-6),1.67-1.56(3H,m,3H's H-2,H-3,H-5,H-6),1.48-1.40(1H,m,1H's H-2,H-3,H-5,H-6),1.17(3H's main product,t,J 7.0Hz,OCH2 C H 3 ), 1.16 (3H minor product, J 7.0 Hz, OCH 2 C H 3 ).

[0306] Example 8

[0307] 2-(1-(trans-4-ethoxycyclohexyl)-4-nitro-1H-pyrazol-3-yl)pyrimidine (E-1)

[0308]

[0309] NaH (60% dispersion in mineral oil, 1.68 g, 42 mmol) was added in portions to a suspension of 2-(4-nitro-1H-pyrazol-3-yl)pyrimidine (D-4, 5.73 g, 30 mmol) in 1,4-dioxane (150 mL, 0.2 M) under a nitrogen atmosphere and cooling in an ice bath. After removing the ice bath, the suspension was stirred at 25 °C. After 3 hours, compound 4-ethoxycyclohexyl 4-nitrobenzenesulfonate (B-5, 11.86 g, 36 mmol, cis / trans ratio ≥ 2) was added, and the reaction mixture was stirred at 100 °C under gentle reflux. After 19 hours, about 0.4 equivalents (5 g) of B-5 were added again, and the reaction was continued. The reaction progress was monitored by LC-MS, and the reaction was stopped on the 4th day. After cooling to room temperature, the reaction was heated to 40 °C with NaHCO 3 The mixture was quenched with saturated aqueous solution (100 mL) and most of the dioxane was removed by rotary evaporation under reduced pressure. The product was extracted with EtOAc (150 mL), which was further washed with brine. The organic layer was dried (Na 2 O) and the mixture was concentrated to 4% by volume. 2 SO 4 ), filtered, and the solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel chromatography, then ground from hexane-EtOAc (8mL-2mL, 35°C, overnight, then collected as a precipitate at room temperature). Compound 2-(1-(trans-4-ethoxycyclohexyl)-4-nitro-1H-pyrazol-3-yl)pyrimidine (E-1) was obtained as an off-white solid: 1.98g (21% yield); 1H NMR (400MHz, chloroform-d) δ8.89(d,J=4.9Hz,2H),8.24(s,1H),7.37(dd,J=4.9,4.9Hz,1H),4.27(tt,J=11.8,3.9Hz,1H),3.55(q,J=7.0Hz,1H),3.35( tt,J=10.7,4.2Hz,1H),2.35–2.29(m,2H),2.26–2.20(m,2H),1.94–1.8 3(m,2H),1.50–1.40(m,2H),1.22(t,J=7.0Hz,3H); LRMS(M+H)m / z318.3.

[0310] Example 9

[0311] 1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-amine

[0312]

[0313] In a Parr flask, 2-(1-(trans-4-ethoxycyclohexyl)-4-nitro-1H-pyrazol-3-yl)pyrimidine (E-1, 974.2 mg, 3.07 mmol) and Pd-BaSO 4 (In BaSO 4 The mixture was stirred for 4 hours at room temperature with 5% Pd (450 mg) in MeOH (30 mL). The reaction was complete as monitored by LC-MS. Under a nitrogen atmosphere, the reaction mixture was passed through a diatomaceous earth pad, which was further washed with MeOH. The filtrate was collected and the solvent was removed in vacuo. Compound 1- (trans-4-ethoxycyclohexyl) -3- (pyrimidin-2-yl) -1H-pyrazole-4-amine was obtained as a light yellow solid (for the next reaction without further purification); 1 HNMR(400MHz, chloroform-d)δ8.75(d,J=4.9Hz,2H),7.09–7.06(m,2H),4.56(br s,2H),4.20(tt,J=12.1,3.7Hz,1H),3.54(q,J=7.0Hz,2H),3.33(tt,J=10.7,4.0Hz,1H),2. 26–2.17(m,4H),1.88–1.78(m,2H),1.47–1.36(m,2H),1.21(t,J=7.0,Hz,3H); LRMS(M+H)m / z 288.3.

[0314] Example 10

[0315] N-(1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol

[0316] -4-yl)oxazole-4-carboxamide (I-1)

[0317]

[0318] 2-(1H-pyrazol-4-yl)thiazole-4-carboxylic acid (599.2 mg, 3.07 mmol) was added to a solution of 1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-amine (3.07 mmol) in DMF (12 mL), cooled in an ice bath, followed by the addition of HATU (1.28 g, 3.38 mmol) after 30 minutes. After another hour, N-ethyl-N-isopropylpropan-2-amine (DIPEA, 1.6 mL, 9.2 mmol) was added dropwise. The ice bath was removed and the mixture was stirred at room temperature for 16 hours. The mixture was stirred for 16 hours by dropwise addition of NaHCO 3 The reaction was quenched with saturated aqueous solution (300 mL), and after 1 h, the precipitate was collected by filtration and washed with H 2 O and then dried in vacuo. A yellow solid (about 1.17 g) was obtained and dissolved in CH 2 Cl 2 –MeOH (10:1). The solution was then passed through a silica gel pad to remove darker impurities. Compound N-(1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol-4-yl)oxazole-4-carboxamide (I-1) was obtained as a light yellow solid: 1.10 g (77% yield); 1 HNMR (400MHz, chloroform-d) δ12.04 (s, 1H), 11.10 (v br s,1H),8.90(d,J=4.9Hz,2H),8.49(s,1H),8.16(s,2H),8.10(s,1H),7.23(dd,J=4.9,4.9Hz,1H),4.34(tt,J=12.0,3.9Hz,1H),3.56(q,J=7 .0Hz,2H),3.37(tt,J=10.8,4.2Hz,1H),2.33–2.29(m,2H),2.25–2.20(m,2H),2.00–1.89(m,2H),1.51–1.41(m,2H),1.22(t,J=7.0Hz,3H); 13C NMR (101 MHz, chloroform-d) δ 161.47, 160.21, 158.35, 157.15, 150.29, 136.25, 132.67, 122.71, 121.86, 119.89, 118.68, 117.15, 76.33, 63.62, 61.83, 31.09, 31.01, 15.65; LRMS (M+H) m / z 465.4.

[0319] The compounds were used without further purification to prepare tartrate salts / co-crystals and for prodrug synthesis. Additionally, if necessary, the compounds were further purified by silica gel chromatography or RP-HPLC.

[0320] Embodiment 11

[0321] Alternative synthesis of I-1

[0322] A. 2-Bromo-N-(1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)thiazole-4-carboxamide (E-2)

[0323]

[0324] 2-Bromothiazole-4-carboxylic acid (298 mg, 1.43 mmol) and HATU (572 mg, 1.50 mmol) were dissolved in CH 2 Cl 2 The solution was stirred at room temperature for 15 minutes (20 mL). N-ethyl-N-isopropylpropan-2-amine (0.623 mL, 3.58 mmol) was added and cooled in an ice bath, followed by the addition of 1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-amine (452 ​​mg, 1.576 mmol) in CH 2 Cl 2 (5 mL) solution. The reaction mixture was stirred at room temperature for 17 hours, after which LC-MS indicated the reaction was complete. By adding NaHCO 3 The reaction was quenched with saturated aqueous solution and stirring was continued for 30 minutes. The two layers were separated and the organic layer was washed again with NaHCO 3 Wash with saturated aqueous solution and 2 SO 4 Dry, filter, and remove the solvent by rotary evaporation under reduced pressure. After silica gel chromatography, compound 2-bromo-N-(1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)thiazole-4-carboxamide (E-2) was obtained as a white solid: 581.1 mg (85% yield); 1H NMR (400MHz, chloroform-d) δ8.89 (d, J = 4.9Hz, 2H), 8.42 (s, 1H), 8.12 (s, 1H), 7.22 (d d,J=4.9,4.9Hz,1H),4.33(tt,J=12.0,3.9Hz,1H),3.56(q,J=7.0Hz,2H),3.3 6(tt,J=10.8,4.2Hz,1H),2.32–2.28(m,2H),2.24–2.20(m,2H),1.99–1.89( m,2H),1.50–1.41(m,2H),1.22(t,J=7.0Hz,3H); LRMS(M+H)m / z477.3,479.3.

[0325] BN-(1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol-4-yl)oxazole-4-carboxamide (I-1)

[0326]

[0327] 2-Bromo-N-(1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)thiazole-4-carboxamide (E-2, 477.4 mg, 1 mmol), (1H-pyrazol-4-yl)boronic acid (279.8 mg, 2.5 mmol), Na 2 CO 3 (318 mg, 3 mmol) and tetrakis(triphenylphosphine)palladium(0) (28 mg, 0.02 mmol) in 1,4-dioxane-H 2 The suspension was degassed with 4 mL of 1% O (4 mL-1 mL) and then backfilled with nitrogen three times. The mixture was heated at 90 °C under a nitrogen atmosphere for 21.5 hours, after which LC-MS indicated the reaction was complete. Most of the dioxane was removed by rotary evaporation under reduced pressure, and the crude product was mixed with water and NaHCO 3 The saturated aqueous solution was mixed until a free-flowing solid appeared. The solid was collected by filtration and washed with H 2 O, and then stirred in hexane-EtOAc-EtOH (1 mL-3 mL-0.5 mL) at 35 ° C within one week. The solid product was collected by filtration, washed with ice-cold hexane-EtOAc (about 1: 1) and dried in vacuo. Compound N-(1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol-4-yl)oxazole-4-carboxamide (I-1) was obtained as a light beige solid: 287.1 mg (62% yield); 1H NMR (400 MHz, chloroform-d) δ 12.04 (s, 1H), 11.10 (v br s,1H),8.90(d,J=4.9Hz,2H),8.49(s,1H),8.16(s,2H),8.10(s,1H),7.2 3(dd,J=4.9,4.9Hz,1H),4.34(tt,J=12.0,3.9Hz,1H),3.56(q,J=7.0Hz,2 H),3.37(tt,J=10.8,4.2Hz,1H),2.33–2.29(m,2H),2.25–2.20(m,2H),2 .00–1.89(m,2H),1.51–1.41(m,2H),1.22(t,J=7.0Hz,3H); LRMS(M+H)m / z 465.4.

[0328] Example 12

[0329] N-(1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol

[0330] -4-yl)thiazole-4-carboxamide (2R,3R)-2,3-dihydroxysuccinic acid

[0331] (I-4; tartrate / co-crystal of I-1)

[0332]

[0333] N-(1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol-4-yl)oxazole-4-carboxamide (I-1, 1.10 g, 2.37 mmol) and (L)-tartaric acid (177.9 mg, 1.19 mmol) were dissolved in CH 2 Cl 2 –MeOH (12 mL–1 mL) solution was stirred at 35 °C for 18 h. 12 mL of CH 2 Cl 2 After cooling to room temperature, the precipitate was collected by filtration and washed with ice-cold CH 2 Cl 2 Washed and dried in vacuo. The product was obtained as a light yellow solid: 702.7 mg; 1 H NMR (400 MHz, DMSO-d 6)δ13.41(br s,1H),12.68(br s,2H),11.93(s,1H),8.98(d,J=4.9Hz,2H),8.52(v br s,1H),8.50(s,1H),8.31(s,1H),8.13(vbr s,1H),7.49(dd,J=4.9,4.9Hz,1H),5.08(br s,2H),4.39–4.31(m,3H),3.50(q,J=7.0Hz,2H),3.41–3.31(m,with H 2 O overlap, 1H), 2.13–2.06 (m, 4H), 1.96–1.86 (m, 2H), 1.41–1.32 (m, 2H), 1.12 (t, J = 7.0 Hz, 3H); 13 C NMR (101 MHz, DMSO-d 6 )δ173.14,161.27,160.61,157.55,157.31,149.07,135.61,132.61,123.08,121.86,1 20.21,119.46,115.64,75.64,72.13,62.58,60.34,30.60,30.50,15.67; LRMS(M+H)m / z 465.5.

[0334] For the filtrate: After removing the solvent, the material was redissolved in CH 2 Cl 2 -MeOH (12 mL-1 mL), and additional (L)-tartaric acid (177.9 mg, 1.19 mmol) was added. After similar reaction and work-up procedures, another batch of product was obtained as a light yellow solid: 280.9 mg, with the same 1 H NMR.

[0335] Combined total: 983.6 mg, 68% yield.

[0336] Example 13

[0337] Di-tert-butyl ((4-(4-((1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)oxazol-2-yl)-1H-pyrazol-1-yl)methyl)phosphate Di-tert-butyl ((4-(4-((1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl)hydrogen phosphate

[0338]

[0339] Cs 2 CO 3 (495 mg, 1.52 mmol) was added to a solution of N-(1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol-4-yl)oxazole-4-carboxamide (I-1, 470.6 mg, 1.01 mmol) in DMF (5 mL) at room temperature. After 10 minutes, di-tert-butyl (chloromethyl)phosphate (524 mg, 2.03 mmol) was added. After 22 hours, the main peak on LC-MS was the desired product (m / z 687.7 (M+H)), leaving <5% of compound I-1. The reaction was quenched by dropwise addition of water (60 mL) while cooling in an ice bath, and stirring was continued at room temperature. After 3 hours, the product was washed with CH 2 Cl 2 (50 mL × 2) extraction. The combined organic layer was dried (Na 2 SO 4 ), filtered, and the solvent removed in vacuo. A light brown oil was obtained and appeared to be mainly mono- t Bu product: 635 mg; 1 HNMR (400MHz, chloroform-d) δ11.97(s,1H),8.88(d,J=4.9Hz,2H),8.01–8.00(m,3H),7.97(s,1 H),7.25(dd,J=4.9,4.9Hz,1H),5.96(d,J=12.1Hz,2H),4.32(tt,J=11.8,3.9Hz,1H),3 .57 (q, J = 7.0 Hz, 2H), 3.37 (tt, J = 10.7, 4.2 Hz, 1H), 2.33–2.30 (m, 2H), 2.25–2.21 (m, 2H), 1.99–1.89 (m, 2H), 1.45 (s, partial overlap, 9H), 1.51–1.36 (m, partial overlap, 2H), 1.23 (t, J = 7.0 Hz, 3H); 31 P NMR (162 MHz, chloroform-d) δ -5.03; LRMS (M+H) m / z 631.6.

[0340] The aqueous layer was adjusted to pH 2 with 1N HCl (aq), and the precipitate was collected and washed with H 2 O was further washed to give 55.3 mg of a bright yellow solid which was shown by LCMS to be mostly acid.

[0341] The di- t Product: 1H NMR (400 MHz, chloroform-d) δ12.08 (s, 1H), 8.96 (d, J = 4.9 Hz, 2H), 8.47 (s, 1H), 8.35 (s, 1H), 8.09 (s, 1H), 8.08 (s, 1H), 7.25 (dd, J = 4.9, 4.9 Hz, 1H), 5.96 (d, J = 13.2 Hz, 2H), 4.34 (tt, J = 11.6, 3.8 Hz, 1H), 3.5 6 (q, J = 7.0 Hz, 2H), 3.37 (tt, J = 10.4, 4.5 Hz, 1H), 2.33-2.30 (m, 2H), 2.24-2.20 (m, 2H), 2.00-1.89 (m, 4H), 1.50-1.41 (m, partial overlap, 2H), 1.44 (s, partial overlap, 18H), 1.22 (t, J = 7.0 Hz, 3H); LRMS (M+H) m / z 687.7. )

[0342] Embodiment 14

[0343] (4-(4-((1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl dihydrogen phosphate (I-2)

[0344]

[0345] Trifluoroacetic acid (TFA) (0.613 mL, 8 mmol) was added to a solution of tert-butyl ((4-(4-((1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl)hydrogen phosphate (635 mg, 1 mmol) in CH 2 Cl 2 The mixture was stirred for 2 hours at room temperature. The volatiles were removed in vacuo and a light brown oil was obtained. The crude product was suspended in acetone-H2O at 35 °C. 2 O (10:1, 12 mL) for 15 h. The solid was collected by filtration, washed with acetone, and further suspended in CH 2 Cl 2 (5 mL) for 1 h. The precipitate was collected by filtration and washed with CH 2 Cl 2Washed and dried in vacuo. Compound (4-(4-((1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl dihydrogen phosphate (I-2) was obtained as a light yellow solid: 277.6 mg (52% yield); 1 H NMR (400 MHz, DMSO-d 6 )δ11.97(s,1H),11.62(v br s,1H),9.02(d,J=4.9Hz,2H),8.63(d,J=0.8Hz,1H),8.49(s,1H),8.36(s,1H), 8.20(d,J=0.8Hz,1H),7.46(dd,J=4.9,4.9Hz,1H),5.92(d,J=11.2Hz,2H),4.3 5(tt,J=11.5,3.7Hz,1H),3.50(q,J=7.0Hz,2H),3.37(tt,J=10.7,3.8Hz,1H), 2.13–2.07(m,4H),1.96–1.85(m,2H),1.41–1.31(m,2H),1.12(t,J=7.0Hz,3H); 31 P NMR (162 MHz, DMSO-d 6 )δ-2.17; LRMS(M+H)m / z575.5.

[0346] Embodiment 15

[0347] Sodium (4-(4-((1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl phosphate (I-3)

[0348]

[0349] 1N NaOH solution (0.9 mL, 0.9 mmol) was added dropwise to a CH 3 CN (2 mL) and H 2O (2 mL) suspension, cooled in an ice bath until pH = 8 is obtained. Stirring is continued for another 10 minutes at room temperature, and the solvent is removed by lyophilization. The compound (4-(4-((1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl sodium phosphate (I-3) is obtained as a cream-colored solid: 278 mg (99% yield); 1 H NMR (400 MHz, deuterium oxide) δ8.63 (d, J = 5.0 Hz, 2H), 8.23 ​​(s, 1H), 8.10 (s, 1H), 7.81 (s, 1H), 7.74 (s, 1H), 7.27 (dd, J = 5.0, 5.0 Hz, 1H), 5.74 (d, J = 6.8 Hz, 2H), 4.21 (br t, J = 12.1 Hz, 1H), 3.73 (q, J = 7.1 Hz, 2H), 3.69–3.59 (m, 1H), 2.31–2.23 (m, 4H), 1.93–1.84 (m, 2H), 1.53–1.44 (m, 2H), 1.25 (t, J = 7.1 Hz, 3H); 31 P NMR (162 MHz, deuterium oxide) δ 2.10; LRMS (M+H) m / z 575.4.

[0350] Example 16

[0351] 1,3-Dihydroxy-2-(hydroxymethyl)propan-2-amine salt (4-(4-((1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-

[0352] Methyl) hydrogen phosphate (I-5)

[0353]

[0354] To (4-(4-((1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl dihydrogen phosphate (I-2, 25 mg, 0.0435 mmol) was added i-PrOH–H 22-Amino-2-(hydroxymethyl)propane-1,3-diol (5.4 mg, 0.044 mmol) was added to the suspension of 1,2-diol-2-(hydroxymethyl)propane-1,3-diol (5.4 mg, 0.044 mmol) in 4% HCl (0.2 mL–0.2 mL), and the dilute suspension was stirred at 30 °C overnight. After cooling to room temperature, additional i-PrOH (0.2 mL, then up to 0.5 mL) was added, and the mixture remained turbid with no free-flowing precipitate. After removing most of the organic solvent by rotary evaporation under reduced pressure, the mixture was resuspended in i-PrOH-ACN-H 2 O until a white solid appears. After filtration, the collected (hygroscopic) solid is dissolved in water and the solvent is removed in vacuo. The title compound 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt (4-(4-((1-(trans-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl hydrogen phosphate (I-5) is obtained as an off-white solid: 15.2 mg (50% yield); 1 H NMR (400MHz, deuterium oxide) δ8.45 (d, J = 4.9Hz, 2H), 8.01 (s, 1H), 7.92 (s, 1H), 7.60 (s, 1H),7.51(s,1H),7.13(dd,J=4.9,4.9Hz,1H),5.68(d,J=7.9Hz,2H),4.11(br t,J=12.2Hz,1H),3.72–3.67(m,8H),3.58(tt,J=11.1,3.9Hz,1H),2.28–2.25(m,2 H),2.20–2.17(m,2H),1.86–1.77(m,2H),1.49–1.40(m,2H),1.23(t,J=7.0Hz,3H); 31 P NMR (162 MHz, deuterium oxide) δ 0.18; LRMS (M+H) m / z 576.1.

[0355] Embodiment 17

[0356] Determination of LPS-induced IL23p19 in THP-1 cells (primed with IFNγ)

[0357] Materials and Equipment

[0358] THP-1 cells (ATCC, catalog number TIB-202), dimethyl sulfoxide (DMSO) (Sigma-Aldrich, catalog number D2650), RPMI 1640 (Cellgro, catalog number 10-040-CM), fetal bovine serum (Sigma, catalog number F4135), albumin from bovine serum (BSA) (Sigma-Aldrich, catalog number A7906), LPS (serotype K-235, Sigma, product number L2143), IFNγ (Peprotech, catalog number 300-02), capture antibody: human IL-23p19 ELISA (e-Bioscience, catalog number 14-7238-85), detection antibody: primary mouse biotinylated anti-human IL-12 (p40 / p70) (e-Bioscience, catalog number 13-7129-85), secondary HRP-conjugated streptavidin (R&D Systems, catalog number DY998), 1x PBST wash buffer (PBS-Tween tablets) (VWR International, catalog number 80058-558), ELISA blocking buffer (PBS with 1% BSA), ELISA dilution buffer (PBS with 1% BSA), 384-well flat bottom, MaxiSorp black immunoplates (Thermo Scientific, catalog number 12-565-346), 384-well flat bottom, white tissue culture plates (ThermoScientific, catalog number 12-565-343), SuperSignal ELISA microchemiluminescent substrate (Thermo Scientific, catalog number 37070), Cell Titer Glo reagent (Promega, catalog number G7573), positive control, IKK2VI inhibitor (Calbiochem, catalog number 401483), AquaMax 4000 plate washer (Molecular Devices), luminometer, WallacVictor2 1420 multilabel counter.

[0359] method

[0360] THP-1 cell stimulation:

[0361] On day 1, 50K / well THP-1 cells were seeded in 384-well plates in RPMI medium with 10% FBS and primed with IFNγ (50 ng / mL) for approximately 18 hours. On day 2, compounds were serially diluted in DMSO in 3-fold dilutions from 5 mM and then diluted 1:125 in RPMI medium with 10% FBS. 50 μL / well 2x compound was added in duplicate to 50 μL / well THP-1 cells (primed with IFNγ) in a 384-well tissue culture plate. Cells were incubated with compounds at 37°C, 5% CO 2 Pre-incubate for 1 hour at 37°C and 5% CO, then add 10 μL / well 11x LPS to give a final concentration of 1 μg / mL LPS. 2 After 18 hours of stimulation, the assay plates were centrifuged and 70 μL / well supernatant was harvested. IL-23p19 protein was measured in 70 μL / well supernatant by sandwich ELISA, and 25 μl / well Cell Titer Glo reagent was added to the remaining cells to measure compound toxicity.

[0362] Human IL-23p19 Sandwich ELISA:

[0363] Maxisorp immune ELISA plates were pre-coated overnight with 25 μL / well anti-IL-23p19 capture antibodies (2.5ug / mL) in PBS at room temperature. After washing with 1x PBST, the plates were blocked with 100 μL / well 1% BSA in PBS for 2 hours at room temperature. The plates were washed three times with 1x PBST and 70 μL / well supernatant was added. The plates were incubated at room temperature with shaking for 2 hours and washed three times with 1x PBST. 25 μL / well biotin-labeled anti-IL-12 (p40 / p70) detection antibodies (100ng / mL) in PBS with 1% BSA were added, and the plates were incubated at room temperature with shaking for 2 hours. After washing three times with 1x PBST, 25 μL / well streptavidin-HRP (1:200) in PBS with 1% BSA was added, and the plates were incubated at room temperature with shaking for 20 minutes. The plate was washed three times with 1xPBST and 25 μL / well of SuperSignal ELISA Micro-chemiluminescent Substrate was added. The plate was read with a luminometer and the chemiluminescent values ​​were entered into Athena (Rigel) for curve fitting, EC 50 Calculation and database storage. The results are shown in Table 1.

[0364] Embodiment 18

[0365] Compound screening using DCs

[0366] Material

[0367] Human PBMC cells (All Cells, catalog number PB002)

[0368] RPMI growth medium containing 10% FBS

[0369] IFNγ (Peprotech, catalog number 300-02)

[0370] GMCSF (Peprotech, catalog number 300-03) and IL4 (Peprotech catalog number 200-04)

[0371] White clear bottom 96-well plate (Fisher, catalog number 07-200-587, Corning #3903)

[0372] LPS (prepared as a 2.5 mg / ml stock solution in PBS) from Sigma-Aldrich (Cat. No. L2018-5MG)

[0373] Cell Titer Glo reagent (Promega, catalog number G7573)

[0374] Positive control, IKK2VI inhibitor (Calbiochem, cat. no. 401483)

[0375] plan

[0376] 1. Differentiation of PBMC to DC cells:

[0377] Human PBMC cells (400 million) obtained from a supplier were transferred to a T-175 flask containing 15 ml of RPMI medium (10% FBS) and incubated for 2 hours at 37° C. After 2 hours, the medium containing the suspended cells was carefully aspirated, and 12 ml of fresh RPMI medium (10% FBS) containing GMCSF (100 ng / ml) and IL4 (20 ng / ml) was added, and the flask was stored in a 37° C. incubator for 7 days.

[0378] After 3 days, fresh GMCSF (100ng / ml) and IL4 (20ng / ml) are added to the flask and continue to incubate. After 7 days, the cells are harvested by rotating (1200rpm / 5min) and sucking off the culture medium to fully differentiate. The cells are suspended in fresh RPMI culture medium (10%FBS) containing 50ng / ml IFNγ (1000U / ml), then plated (50K / well, in 100μl) in white transparent bottom 96-well plates and placed in 37°C incubators for 24 hours.

[0379] 2. Addition of compounds:

[0380] After 24 hours of incubation, 100 μl of RPMI medium containing 2X concentrated test compounds per well was added to the above cell culture medium (final concentration became 1X), and the plate was pre-incubated at 37°C for 1 hour before stimulation with LPS.

[0381] After 1 hour of compound pre-incubation, 10 μl per well of 20X concentrated LPS solution in RPMI medium was added to give a final concentration of 1 μg / ml. The mixture was shaken and the plate was incubated at 37°C for another 18 hours.

[0382] Carefully harvest 155 μl of supernatant from each well (without touching the bottom of the well with a tip), and add 50 μl of Cell Titer Glo reagent to the remaining 50 μl / well of the cell culture plate. The mixture is incubated on a shaker for 1-2 minutes, and the luminescence intensity of the plate is read to determine the compound cytotoxicity. The cell culture supernatant collected above is used to perform IL23 ELISA (65 μl-supernatant) and IL10 ELISA (90 μl-supernatant) as described below.

[0383] Embodiment 19

[0384] Human IL-23 (p19 / p40) ELISA protocol (e-Bioscience)

[0385] Material:

[0386] 96-well high binding opaque white plates (from Pierce, catalog number 15042);

[0387] 1X PBS; 1X TBST wash buffer;

[0388] Blocking solution: 0.5% casein (from BDH, catalog number 440203H) in PBS;

[0389] Dilution solution: 1% BSA in PBS (10% BSA from Fisher, catalog number 37525);

[0390] Capture antibody: rat anti-human IL-23 (p19) (e-Bioscience, catalog number 14-7238-85);

[0391] Detection antibody: primary mouse biotinylated anti-human IL-12 (p40 / p70) (e-bioscience, catalog number 13-7129-85);

[0392] secondary HRP-conjugated streptavidin (R&D Systems, catalog number DY998);

[0393] r Human-IL-23 (e-bioscience, catalog number 34-8239) (recommended starting concentration = 5 ng / ml in RPMI cell culture medium);

[0394] cell culture supernatant (65 μl from THP-1 cells primed with IFNγ (50 ng / ml–1000 U / ml) and stimulated with 0.01% SAC);

[0395] SuperSignal ELISA Microchemiluminescent Substrate [Pierce, Cat. No. 37069].

[0396] Coated board:

[0397] Capture antibody (2.5 μg / ml) was added to 10.5 ml PBS to which 50 μl of anti-IL23 (p19) was added. The mixture was mixed thoroughly and 100 μl of coating solution was added to each well of a 96-well white plate from Pierce. The wells were covered and incubated overnight at 4°C.

[0398] Closed plate:

[0399] Anti-IL23(p19)-antibody coated plates were washed 2X with TBST (using a plate washer) and blocked with 200 μl of 0.5% casein for 1.5-2 hours at room temperature with shaking.

[0400] Add supernatant and detect:

[0401] The plates were washed 2X with TBST and the supernatants (65 μl / well) were transferred to the above pre-blocked / IL23(p19)-antibody coated 96-well plates and incubated at room temperature for 1.5 hours with shaking.

[0402] The plate was washed 4X with TBST (plate washer) and 100 μl / well detection antibody solution prepared from 2 μl biotinylated anti-IL-12 (p40 / p70) antibody in 11 ml 1% BSA / PBS solution (1-5000 dilution) was added. The plate was incubated at room temperature for 1 hour with shaking.

[0403] Again, the plate was washed 4X with TBST, and 100 μl of HRP-labeled streptavidin (R&D Systems) solution (10 μl / 10 ml 1% BSA solution) was added, and the plate was incubated for another 45 minutes at room temperature with shaking.

[0404] After 45 minutes, the plates were washed 4X with TBST and 100ul / well SuperSignal ELISA Mini-Chemiluminescent Substrate from Pierce (3.5ml A+3.5ml B+3.5ml MQ water) was added. The plates were shaken for 1-2 minutes and then read on a plate reader.

[0405] Embodiment 20

[0406] IRAK4 ADP-Glo ​​assay

[0407] Material

[0408] IRAK4 kinase (Signalchem, I12-10G-20); 0.1M DTT (Signalchem, D86-09B); MBP substrate (Signalchem, M42-51N); ADP Glo (Promega, V9101); 1M MgCl 2 (Teknova, M03304); 1M Tris-HCl pH 7.4 (Teknova, T1074); BSA (Sigma (A3059)); distilled H 2 O

[0409] equipment

[0410] Wallac Victor2 1420 Multi-Label Counter

[0411] method

[0412] ADP-Glo TM Thaw reagents at ambient temperature. Prepare kinase assay reagents by mixing kinase assay buffer with lyophilized kinase assay substrate and set aside.

[0413] In distilled HO with a final pH of 7.4 2 O with a final concentration of 100 mM MgCl 2 Prepare a stock volume of 5X reaction kinase buffer with 200 mM Tris-HCl and 0.5 mg / ml BSA. Prepare a 2X working stock volume of reaction kinase buffer to a final concentration of 100 μM DTT.

[0414] Thaw the components of IRAK4 enzyme on ice. Prepare diluted IRAK4 at 5.0 ng / μl in 1X kinase reaction buffer (diluted from 2X buffer). Prepare 250 μM working stock ATP assay solution in 1X kinase reaction buffer (diluted from 2X buffer).

[0415] Compounds were diluted 8 points in 4-fold serial dilutions from 250 μM in DMSO. Then diluted 1:5 in 2X reaction buffer in a 96-well plate. 1.0 μl was transferred to a 384-well plate in duplicate. 2 μl of diluted active IRAK4 (not added to column 1) was added, and 2X reaction buffer was added to column 1. 1 μl of 1 mg / ml MBP stock solution was added. It should be noted that MBP can be combined with an equal volume of ATP and then added at 2 μl / well. The final reaction volume was 5 μl.

[0416] The plates were centrifuged and the reaction mixtures were incubated at room temperature for 60 minutes or at 30°C for 30 minutes.

[0417] The reaction was stopped by adding 5 μl of ADP-Glo TM The 384-well plate was centrifuged and the reaction mixture was then incubated at ambient temperature for another 40 minutes.

[0418] 10 μl of kinase detection reagent was added. The plate was centrifuged and the reaction mixture was then incubated for a further 30 minutes at ambient temperature.

[0419] The 384-well reaction plates were read using a WALLAC plate reader (luminescence 0.1 s).

[0420] Embodiment 21

[0421] IRAK1 ADAPTA assay

[0422] Material

[0423] Barcoded Corning Low Volume White 384-well Plate (Corning Catalog No. 4512)

[0424] - Test compounds: Test compounds were screened in 1% DMSO (final) in wells. For 10 point titration, 3-fold serial dilution, 10 μM.

[0425] Substrate / kinase mix: in 50 mM HEPES pH 7.5, 0.01% BRIJ-35, 10 mM MgCl 2 2X IRAK1 / histone H3 (1-20) peptide mixture was prepared in 1 mM EGTA.

[0426] ATP solution: Dilute all ATP solutions in water to 4X working concentration.

[0427] • The apparent ATP Km was previously determined using a radioactive assay, except when no substrate was available, in which case the Adapta assay was performed.

[0428] Detection mixture: Prepare the detection mixture in TR-FRET dilution buffer. The detection mixture consists of EDTA (30 mM), Eu-anti-ADP antibody (6 nM) and ADP tracer. The detection mixture contains EC 60 Concentrations of tracer were used ranging from 5-150 μM ATP.

[0429] IRAK1 approach

[0430] 1.100 nL – 100X test compound in 100% DMSO

[0431] 2.2.4 μL – 30 mM HEPES

[0432] 3.2.5 μL – 4X ATP solution

[0433] 4.5 μL – 2X Substrate / Kinase Mix

[0434] The final 10 μL kinase reaction consisted of 32.5 mM HEPES pH 7.5, 0.005% BRIJ-35, 5 mM MgCl 2 , 3.17-42 ng IRAK1 and 100 μM histone H3 (1-20) in 0.5 mM EGTA.

[0435] 5.30 seconds plate shaking

[0436] 6. Centrifuge at 1000 x g for 1 minute

[0437] 7. Incubate the kinase reaction at room temperature for 60 minutes

[0438] 8.5 μL – Detection mixture

[0439] 9.30 seconds plate shaking

[0440] 10. Centrifuge at 1000 x g for 1 minute

[0441] 11. Allow the mixture to equilibrate at room temperature for 60 minutes

[0442] 12. Read and analyze data on a fluorescence plate reader

[0443] Embodiment 22

[0444] IRAK4 Z'-dielectric determination

[0445] Material

[0446] Barcoded Corning Low Volume NBS Black 384-well Plates (Corning Cat. No. 4514)

[0447] - Test compounds: Test compounds were screened in wells in 1% DMSO (final). For a 10 point titration, 3-fold serial dilutions were performed from 10 μM.

[0448] Peptide / kinase mixture: in 50 mM HEPES pH 7.5, 0.01% BRIJ-35, 10 mM MnCl 2 , 1mMEGTA, 2mM DTT, 0.02%NaN 3 Prepare 2X IRAK4 / Ser / Thr 07 mixture in .

[0449] ATP solution: in kinase buffer (50 mM HEPES pH 7.5, 0.01% BRIJ-35, 10 mM MgCl 2 All ATP solutions were diluted to 4X working concentration in 1 mM EGTA. The assay determines the apparent ATP Km.

[0450] • Developer Solution: Dilute the developer 1:45,000 in developer buffer.

[0451] General approach

[0452] 1.100 nL – 100X test compound in 100% DMSO

[0453] 2.2.4 μL – Kinase buffer

[0454] 3.5 μL – 2X Peptide / Kinase Mix

[0455] 4.2.5 μL – 4X ATP solution

[0456] The final 10 μL kinase reaction consisted of 50 mM HEPES pH 7.5, 0.01% BRIJ-35, 5 mM MgCl 2 , 5mM MnCl 2 , 1mM EGTA, 1mM DTT, 0.01% NaN 3 The composition was 3.45-63.6ng IRAK4 and 2μM Ser / Thr 07.

[0457] 5.30 seconds plate shaking

[0458] 6. Incubate the kinase reaction at room temperature for 60 minutes

[0459] 7.5μL – developer solution

[0460] 8.30 seconds plate shaking

[0461] 9. Incubate the development reaction at room temperature for 60 minutes

[0462] 10. Read and analyze data on a fluorescence plate reader

[0463] The results from the assay are shown in Table 1.

[0464] Table 1. In vitro results.

[0465]

[0466] Embodiment 23

[0467] Plasma protein binding of I-1

[0468] Experimental Procedure

[0469] Brain and plasma drug binding was performed by equilibrium dialysis using a rapid equilibrium dialysis (RED) device. Briefly, 300 μL plasma or brain homogenates (1 brain / 3 buffers) containing drug (5 μM) were balanced against 500 μL buffered saline (4-18 hours). After incubation, the concentration of the drug in plasma / brain homogenates and buffer was determined by LC-MS / MS.

[0470] Recovery value

[0471] Recovery was calculated by determining the total amount of drug in 50 μL plasma and 50 μL buffer after incubation and comparing this value to the amount of drug in a 50 μL plasma sample that was not incubated. For protein binding assays, drug was added to plasma and this plasma was added to one side of the dialysis device while buffer was added to the other side. After submission to the plasma chamber of the equilibration device, a 50 μL aliquot of plasma was mixed with 50 μL buffer and then quenched with a stop reagent.

[0472] Drug recovery = (drug buffer + drug plasma) / (drug in initial plasma)

[0473] The lower recovery may be due to hydrolysis of drug in plasma that was lost due to binding to the device (ie, amide hydrolysis).

[0474] Free drug

[0475] Higher free drug in plasma and brain allows more drug to interact with the target.

[0476]

[0477] From the perspective of free drug, the freeness of Compound I-1 in rat, dog, monkey and human proteins is about 6 times that of Comparative-1.

[0478] The mouse recovery rate was lower due to hydrolysis, which is comparable to other similar experiments in mice.

[0479] Table 2. Plasma protein binding results

[0480]

[0481] *Due to experimental error, results may appear to be greater than 100%.

[0482] Two rounds of rat brain homogenate bound to 1-1 compared to Comparative-1. The free degree of 1-1 in rat brain was approximately 10 times that of Comparative-1.

[0483] Comparative-1 0.102% and 0.114%

[0484] I-1 1.06% and 1.43%

[0485] Embodiment 24

[0486] Whole blood assay

[0487] Compound 1-1 exhibited similar biochemical potency to Comparator-1 in in vitro assays (see Example 23 for the structure of Comparator-1), but 1-1 was significantly more potent in assays performed in cells and in the presence of whole blood.

[0488] Table 3. In vitro assays

[0489]

[0490] Table 4. Free fraction in whole blood assay

[0491] Compound Rat Human I-1 3.7％ 3.0％ Comparison-1 0.7％ 0.4％

[0492] The greater potency of 1-1 in the whole blood assay was driven by the plasma free fraction, and the free fraction percentage was consistent between species.

[0493] To evaluate the effects of Compound 1-1 on TLR- and IL-1R-dependent cytokine release, various cell types were isolated and stimulated in the presence of Compound 1-1 or Comparator-1. The results are provided in Table 5.

[0494] Table 5. Cell assays

[0495]

[0496] The results obtained in the above cell assays show that compound I-1 is an effective inhibitor of TLR and IL-1R dependent cytokine production in various cell types (Table 5). Compound I-1 inhibits TLR4-induced IL-23 and TNF-α production by human THP-1 cells. In human primary dendritic cells (DCs), compound I-1 blocks TLR4-induced IL-23 production, TLR7 and TLR2-induced TNF-α production. Compound I-1 is also an effective inhibitor of TLR7-dependent cytokine production in human PBMCs and TLR4-dependent cytokine production in mouse macrophages. In addition, compound I-1 effectively inhibits IL-1β-induced cytokine IL-6 production in primary human endothelial cells. Compound I-1 widely inhibits TLR and IL-1R-dependent proinflammatory cytokine production in various primary human cell types by inhibiting IRAK1 / 4 kinase activity. In addition, compound I-1 is a more effective inhibitor of TLR and IL-1R-dependent signal transduction in various cell types.

[0497] Assay protocol: TLR4- and TLR2-induced cytokines in THP-1 cells.

[0498] Material

[0499] Dimethyl sulfoxide (DMSO) (Sigma-Aldrich, catalog number D2650)

[0500] Fetal bovine serum (Sigma, catalog number F4135)

[0501] RPMI 1640 (Cellgro, catalog number 10-040-CM)

[0502] THP-1 (ATCC, catalog number TIB-202)

[0503] Recombinant human IFN-γ (Peprotech, catalog number AF-300-02)

[0504] Lipopolysaccharide (LPS) from Escherichia coli K-235 (Sigma-Aldrich, catalog number L2018)

[0505] LTA (InvivoGen, catalog number tlrl-pslta)

[0506] Anti-human IL-23p19 (eBioscience, catalog number 14-7238-85)

[0507] Biotin anti-human IL-12 (p40 / 70) (eBioscience, catalog number 13-7129-85)

[0508] Bovine serum albumin (BSA) (Sigma-Aldrich, catalog number A7906)

[0509] 1x PBST Wash Buffer (PBS-Tween tablets) (VWR International, catalog number 80058-558)

[0510] ELISA blocking buffer (PBS with 2% BSA)

[0511] ELISA dilution buffer (0.2% BSA in PBST)

[0512] Human TNF-α DuoSet ELISA Kit (R&D Systems, catalog number DY210)

[0513] · Luminescent Cell Viability Assay (Promega, catalog number G7571)

[0514] 96-well flat-bottom white tissue culture plates with clear bottom (Costar 3903)

[0515] 96-well flat bottom MaxiSorp black immunoplate (Thermo Scientific, catalog number 12-566-24)

[0516] SuperSignal ELISA Micro-Chemiluminescent Substrate (Thermo Scientific, Cat. No. 37070)

[0517] equipment

[0518] Scan Washer 400 (Molecular Devices)

[0519] Photometer, Wallac Victor2 1420 multi-label counter

[0520] method

[0521] THP-1 cells (100K, 100 μL / well) were cultured at 37°C and 5% CO 2 Compounds were serially diluted in DMSO at 4-fold dilutions from 2.5 mM and then diluted 1:125 in RPMI 1640 complete medium with 10% FBS. 100 μL / well 2x compound was mixed with 100 μL / well IFN-γ primed THP-1 in a 96-well flat-bottom white tissue culture plate. Cells were incubated with compound at 37°C, 5% CO 2After pre-incubation for 1 hour at 37°C and 5% CO, 10 μL / well of 21x LPS was added to give a final concentration of 1 μg / mL LPS. 2 After centrifugation, 100 μL / well of supernatant was extracted by using a multichannel pipette. IL-23p19 or TNF-α protein in the supernatant was measured by sandwich ELISA. Cell viability assays were performed using the remaining IFN-γ-induced THP-1 cell pellets to determine compound cytotoxicity. The plates were read with a photometer and the chemiluminescence values ​​were entered into Athena (Rigel) for curve fitting, EC 50 Computing and database storage.

[0522] THP1 cells primed with IFN-γ as described previously were cultured at 37°C, 5% CO 2 The cells were restimulated for 18 hours with LTA (10 μl / well of 20x concentrated LTA solution in RPMI medium to give a final concentration of 4 μg / ml). The supernatant (155 μl / well) was harvested to measure TNF-α. TNF-α was quantified by ELISA using the Human TNF-α DuoSet ELISA Kit according to the kit protocol.

[0523] Human IL-23p19 Sandwich ELISA:

[0524] Maxisorp immune ELISA plates were pre-coated overnight with 100 μL / well anti-IL-23p19 capture antibodies (2.4 μg / mL) in PBS at room temperature. After washing with 1x PBST, the plates were blocked with 300 μL / well 1% BSA in PBS at room temperature for 2 hours. The plates were washed three times with 1x PBST, and 100 μL / well supernatant was added. The plates were incubated at room temperature for 2 hours while shaking. The plates were washed three times with 1x PBST, and 100 μL / well biotin anti-human IL-12 (p40 / 70) detection antibodies (10 ng / mL) were added in PBS with 0.1% BSA. The plates were incubated at room temperature for 2 hours while shaking. The plates were washed three times with 1x PBST, and 100 μL / well streptavidin-HRP (1:200) was added in PBS with 0.1% BSA. The plates were incubated at room temperature for 20 minutes while shaking. The plates were washed three times with 1x PBST and 100 μL / well of SuperSignal ELISA Micro-chemiluminescent Substrate was added. The plates were read with a luminometer and the chemiluminescent values ​​were entered into Athena (Rigel) for curve fitting, EC 50 Computing and database storage.

[0525] Assay protocol: TLR7-induced TNF-α in human peripheral blood mononuclear cells (PBMCs).

[0526] Material

[0527] Corning TM Costar TM 96-well cell culture treated U-bottom microplate (Fisher, catalog number 07-200-95)

[0528] Human PBMC cells (ALL Cells, catalog number PB002)

[0529] Resiquimod (InvivoGen, catalog number tlrl-r848)

[0530] Human TNF-α DuoSet ELISA Kit (R&D Systems, catalog number DY210)

[0531] Recombinant human IFN-β (Peprotech, catalog number 300-02BC)

[0532] RPMI with 10% FBS

[0533] Axygen 96-well 1.1 mL sterile (VWR International, catalog number 47734-788)

[0534] method

[0535] Human peripheral blood mononuclear cells (PBMCs) were seeded at 100,000 cells / well in 100 μl of RPMI with 10% FCS and incubated at 37°C, 5% CO 2 The compounds were serially diluted in DMSO in 3-fold dilutions from 2 mM and then diluted 1:100 in RPMI 1640 complete medium with 10% FBS (2x stock solution). PBMCs were incubated at 37°C, 5% CO by adding 100 μl RPMI medium containing 2x concentrated test compound / well (0.5% final DMSO concentration). 2 Preincubate with test compounds for 1 hour.

[0536] Resiquimod irritation:

[0537] PBMCs were cultured at 37°C and 5% CO 2The cells were restimulated for 18 hours with resiquimod (10 μl / well of 21× concentrated resiquimod solution in RPMI medium to give a final concentration of 0.1 μg / ml). 100 μl of supernatant / well was harvested to measure TNF-α. TNF-α was quantified using the DuoSet ELISA kit according to the manufacturer's protocol.

[0538] Assay Protocol: TLR-induced cytokines in human primary monocyte-derived dendritic cells (DCs).

[0539] Material

[0540] Dimethyl sulfoxide (DMSO) (Sigma-Aldrich, catalog number D2650)

[0541] Fetal bovine serum (Sigma, catalog number F4135)

[0542] Human PBMC cells (All Cells, catalog number PB002)

[0543] RPMI growth medium containing 10% FBS

[0544] IFN-γ (Peprotech, catalog number 300-02)

[0545] GMCSF (Peprotech, catalog number 300-03) and IL4 (Peprotech, catalog number 200-04)

[0546] White clear bottom 96-well plate (Fisher, catalog number 07-200-587, Corning #3903)

[0547] LPS (prepared as a 2.5 mg / ml stock solution in PBS) from Sigma-Aldrich (Cat. No. L2018-5MG)

[0548] Lipopolysaccharide (LPS) from Escherichia coli K-235 (Sigma-Aldrich, catalog number L2018)

[0549] Gadmot, a new imidazoquinoline compound (InvivoGen, catalog number tlrl-gdqs)

[0550] Anti-human IL-23p19 (eBioscience, catalog number 14-7238-85)

[0551] Biotin anti-human IL-12 (p40 / 70) (eBioscience, catalog number 13-7129-85)

[0552] Bovine serum albumin (BSA) (Sigma-Aldrich, catalog number A7906)

[0553] 1x PBST Wash Buffer (PBS-Tween tablets) (VWR International, catalog number 80058-558)

[0554] ELISA blocking buffer (PBS with 2% BSA)

[0555] ELISA dilution buffer (0.2% BSA in PBST)

[0556] Human TNF-α DuoSet ELISA Kit (R&D Systems, catalog number DY210)

[0557] Cell Titer Glo reagent (Promega, catalog number G7573)

[0558] Positive control, IKK2VI inhibitor (Calbiochem, catalog number 401483)

[0559] equipment

[0560] Scan Washer 400 (Molecular Devices)

[0561] Photometer, Wallac Victor2 1420 multi-label counter

[0562] method

[0563] Dendritic cell differentiation and IFNγ priming:

[0564] Human peripheral blood mononuclear cells (PBMC) (400 million) were cultured at 37°C and 5% CO 2 Monocytes were captured by allowing them to attach for 2 hours in a T-175 flask containing 15 ml of RPMI medium (10% FBS) at 4 °C.

[0565] After 2 hours, the culture medium containing the suspended cells was carefully removed, and 12 ml of fresh RPMI medium (10% FBS) containing GM-CSF (100 ng / ml) and IL-4 (20 ng / ml) was added to the attached monocytes. The monocytes were incubated at 37°C, 5% CO 2 The cells were differentiated for 7 days and fresh GM-CSF and IL-4 were added after 3 days.

[0566] Differentiated dendritic cells (DCs) were harvested by centrifugation (1200 rpm / 5 min). DCs were cultured in a white clear-bottom 96-well plate at 37°C and 5% CO. 2 The cells were primed with 50 ng / ml IFN-γ (1000 U / ml) at 50K / well in 100 μl / well of fresh RPMI medium (10% FBS) for 24 hours.

[0567] Compound pre-incubation:

[0568] Compounds were serially diluted in DMSO in 4-fold dilutions from 2.5 mM and then diluted 1:125 in complete RPMI1640 medium with 10% FBS. Primed DCs were incubated at 37°C, 5% CO by adding 100 μl RPMI medium containing 2X concentrated test compound / well (0.5% final DMSO concentration). 2 Preincubate with test compounds for 1 hour.

[0569] LPS stimulation:

[0570] DCs were maintained at 37°C and 5% CO 2 The cells were restimulated with LPS (10 μl / well of 20x concentrated LPS solution in RPMI medium to give a final concentration of 1 μg / ml) for 18 h.

[0571] The supernatant (155 μl / well) was harvested to measure IL-23 by ELISA (65 μl supernatant). Cell Titer Glo reagent (50 μl / well) was added to the remaining 50 μl / well of the cell culture plate, incubated on a shaker for 1-2 minutes, and the luminescence intensity of the plate was read to determine compound cytotoxicity. The chemiluminescence values ​​were entered into Athena (Rigel) for curve fitting, EC 50 Computing and database storage.

[0572] Gadmott Stimulus:

[0573] DCs were maintained at 37°C and 5% CO 2 The cells were restimulated for 18 hours with galdemoline (10 μl / well of 20× concentrated galdemoline solution in RPMI medium to give a final concentration of 4 μg / ml). The supernatant (155 μl / well) was harvested to measure TNF-α. TNF-α was quantified by ELISA according to the kit protocol.

[0574] Whole blood assay:

[0575] In order to evaluate the effect of whole blood on the efficacy of compound I-1 and comparator-1, the inhibitory potential of the compound for TLR-dependent cytokine release in human blood was evaluated. Peripheral blood freshly drawn from healthy volunteers was incubated with the test compound and stimulated overnight with LPS or gadmote. Serum was separated after centrifugation, and TNF-α and IL-6 were measured by ELISA. The results are shown in Table 6.

[0576] Assay Protocol: LPS-induced TNF-α in human blood.

[0577] Material

[0578] RPMI 1640 (Cellgro, catalog number 10-040-CM)

[0579] Fetal bovine serum (Sigma, catalog number F4135)

[0580] Lipopolysaccharide from Escherichia coli K-235 (Sigma-Aldrich Chemicals Cat. No. L2018-5MG)

[0581] BD Vacutainer Heparin Sodium Tube (BD Catalog No. 4191982)

[0582] 96-well polystyrene plates (Fisher Cat. No. 12-565-500)

[0583] Dimethyl sulfoxide (DMSO) (Sigma-Aldrich, catalog number D2650)

[0584] Human cytokine magnetic plate: Magnetic (EMD Millipore Corp., catalog number HCYTOMAG-60K-02)

[0585] equipment

[0586] Luminex Flexmap 3d

[0587] method

[0588] Whole blood from healthy volunteers was collected in a Vacutainer containing sodium heparin and 40 μl of blood was plated into each well. Compounds were serially diluted in DMSO at 3-fold dilutions from 2 mM and then diluted 1:100 in medium with 10% FBS. 50 μL of 2x compound per well was added in duplicate to each well and incubated for one hour. 10 μL of LPS was added to each well at a final concentration of 0.5 ng / ml and the cells were incubated overnight.

[0589] 100 μL of PBS was added to each well and centrifuged at 1000 RPM for 10 minutes. 25 μL of supernatant was taken and TNF-α was measured on Luminex Flexmap 3d.

[0590] Assay protocol: Gadmoxetine-induced IL-6 in human blood, TNF-α in human blood.

[0591] Material

[0592] RPMI 1640 (Cellgro, catalog number 10-040-CM)

[0593] Fetal bovine serum (Sigma, catalog number F4135)

[0594] Gadmot (InvivoGen catalog number tlrl-gdqs)

[0595] BD Vacutainer Heparin Sodium Tube (BD Catalog No. 4191982)

[0596] 96-well polystyrene plates (Fisher Cat. No. 12-565-500)

[0597] Dimethyl sulfoxide (DMSO) (Sigma-Aldrich, catalog number D2650)

[0598] Human cytokine magnetic plate: Il6 Magnetic (EMD Millipore Corp. catalog number HCYTOMAG-60K-02)

[0599] equipment

[0600] Luminex Flexmap 3d

[0601] method

[0602] Whole blood from healthy volunteers was collected in a Vacutainer containing sodium heparin and 40 μl of blood was plated into each well. Compounds were serially diluted in DMSO at 3-fold dilutions from 2 mM and then diluted 1:100 in medium with 10% FBS. 50 μL of 2X compound per well was added in duplicate to each well and incubated for one hour. 10 μL of Gadmot was added to each well at a final concentration of 4 μg / ml and the cells were incubated overnight.

[0603] 100 μL of PBS was added to each well and centrifuged at 1000 RPM for 10 minutes. 25 μL of supernatant was taken and IL-6 was measured by Luminex Flexmap 3d.

[0604] Table 6. Human whole blood assay

[0605]

[0606] Compound 1-1 exhibited greater potency in blocking inflammatory signaling in a cell-based assay and lost less potency than Comparator-1 when tested in the presence of human whole blood.

[0607] Embodiment 25

[0608] Prodrug Compound I-3

[0609] Table 7. Bioavailability of Compound I-3

[0610] Species Bioavailability of compound I-3 (%F) Mouse 34 Rat 12 monkey 71 dog 53

[0611] The bioavailability of Compound 1-3 was measured as the amount of parent Compound 1-1 detected, demonstrating that Compound 1-3 acts as a prodrug of 1-1 in vivo.

[0612] In view of the many possible embodiments to which the principles of the disclosed technology may be applied, it should be recognized that the embodiments shown are merely preferred embodiments of the technology and should not be considered to limit the scope of the technology. Rather, the scope of the technology is defined by the appended claims. Therefore, we claim that our technology falls entirely within the scope and spirit of these claims.

Claims

1. A compound having a structure according to formula I or a pharmaceutically acceptable salt or solvate thereof, wherein: R 1 is H, aliphatic, acyl, heterocyclic, carboxyl ester, amide, alkyl phosphoramidate or alkyl phosphate; R 2 C 1-6 alkyl.

2. The compound of claim 1, wherein R 1 is H, alkyl or -alkylOP(O)(OR)2, wherein each OR is -OH, -Oalkyl, -Oaryl, -Oheteroaryl, -Oaralkyl or -O - M + , where M + It is a counter ion with a single positive charge.

3. The compound of claim 1 or claim 2, wherein R 1 It is -CH(CH3)OP(O)(OR)2 or -CH2OP(O)(OR)2.

4. A compound as described in any one of claims 1 to 3, wherein R 2 C 1-4 alkyl.

5. A compound as described in any one of claims 1 to 4, wherein the compound has a structure according to Formula II or a pharmaceutically acceptable salt or solvate thereof.

6. A compound as described in any one of claims 1 to 5, wherein R 1 For H.

7. A compound as described in any one of claims 1 to 5, wherein R 1 C 1-6 alkyl.

8. A compound as described in any one of claims 1 to 5, wherein R 1 It is -CH2OP(O)(OR)2.

9. The compound of claim 8, wherein R 1 It is -CH2OP(O)(OH)2.

10. The compound of claim 8, wherein R 1 -CH2OP(O)(OC 1-6 Alkyl)2.

11. The compound of claim 8, wherein R 1 -CH2OP(O)(O - M + )2.

12. The compound of claim 8, wherein R 1 -CH2OP(O)(O - Na + )2.

13. The compound of any one of claims 1-12, wherein the compound is in free base form.

14. The compound of any one of claims 1-12, wherein the compound is a salt co-crystal.

15. The compound of claim 14, wherein the salt co-crystal is a tartrate co-crystal or a triple salt co-crystal.

16. The compound of claim 1, wherein the compound is selected from: I-1: N-(1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol-4-yl)thiazole-4-carboxamide; I-2: (4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl dihydrogen phosphate; I-3: sodium (4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl phosphate; I-4: N-(1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)-2-(1H-pyrazol-4-yl)thiazole-4-carboxamide tartrate; I-5: (4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)methyl hydrogen phosphate 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt (trisalt); I-6: 1-(4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)ethyl dihydrogen phosphate; I-7: sodium 1-(4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)ethyl phosphate; or I-8: 1-(4-(4-((1-((1r,4r)-4-ethoxycyclohexyl)-3-(pyrimidin-2-yl)-1H-pyrazol-4-yl)carbamoyl)thiazol-2-yl)-1H-pyrazol-1-yl)ethyl hydrogen phosphate 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt (trisalt).

17. A pharmaceutical composition comprising the compound of any one of claims 1 to 16 and a pharmaceutically acceptable excipient.

18. A method for inhibiting an IRAK enzyme, comprising contacting the enzyme with an effective amount of a compound according to any one of claims 1 to 16.

19. The method of claim 18, wherein contacting comprises administering the compound to the subject.

20. A method for treating a disease or condition in a subject requiring an IRAK inhibitor, the method comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 16 or a pharmaceutical composition according to claim 17.

21. The method of claim 20, wherein the disease or disorder comprises an autoimmune disease, an inflammatory disorder, a cardiovascular disease, a neurodegenerative disorder, an allergic disorder, multiple organ failure, kidney disease, platelet aggregation, a hyperproliferative disorder, transplantation, sperm motility, red blood cell deficiency, transplant rejection, lung injury, respiratory disease, ischemic disease, bacterial infection, viral infection, immunomodulatory disorder, sickle cell disease, chemical or radiation induced lung injury, hemorrhagic fever, or a combination thereof.

22. The method of claim 20, wherein the disease or condition comprises aplastic anemia, atopic dermatitis, pustular psoriasis, palmoplantar pustulosis, primary biliary cirrhosis, pyoderma, sclerosing cholangitis, systemic juvenile idiopathic arthritis, hidradenitis suppurativa, cytokine release syndrome, or myelodysplastic syndrome (MDS).

23. The method of claim 20, wherein the disease is amyotrophic lateral sclerosis (ALS), systemic lupus erythematosus, chronic rheumatoid arthritis, type I diabetes, inflammatory bowel disease, biliary cirrhosis, uveitis, multiple sclerosis, Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, psoriasis, autoimmune myositis, pancreatitis, Kaposi's sarcoma, myelodysplastic syndrome, Wegener's granulomatosis, ichthyosis, Graves' ophthalmopathy, or asthma.

24. The method of claim 21, wherein the immunomodulatory disorder is rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, systemic sclerosis, myasthenia gravis, type I diabetes, uveitis, posterior uveitis, allergic encephalomyelitis, glomerulonephritis, post-infectious autoimmune diseases including rheumatic fever and post-infectious glomerulonephritis, inflammatory and hyperproliferative skin diseases, psoriasis, atopic dermatitis, contact dermatitis, eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermolysis bullosa, urticaria, angioedema, vasculitis, erythema, cutaneous eosinophilia, lupus erythematosus, acne, alopecia areata, keratoconjunctivitis, vernal conjunctivitis, and Uveitis associated with Behcet's disease, keratitis, herpetic keratitis, keratoconus, corneal epithelial dystrophy, corneal leukoplakia, pemphigus vulvae, Mooren's ulcer, scleritis, Graves' ophthalmopathy, Vogt-Koyanagi-Harada syndrome, sarcoidosis, pollen allergy, reversible obstructive airway disease, bronchial asthma, allergic asthma, intrinsic asthma, exogenous asthma, dust asthma, chronic or deep-rooted asthma, advanced asthma and airway hyperresponsiveness, bronchitis, gastric ulcer, vascular damage caused by ischemic disease and thrombosis, ischemic bowel disease, inflammatory bowel disease, necrotizing enterocolitis, intestinal lesions associated with thermal burns, celiac disease, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease, ulcerative colitis, Migraine, rhinitis, eczema, interstitial nephritis, Goodpasture's syndrome, hemolytic uremic syndrome, diabetic nephropathy, polymyositis, Guillain-Barré syndrome, Meniere's disease, polyneuritis, polyneuritis, mononeuritis, radiculopathy, hyperthyroidism, Basedow's disease, pure red cell aplasia, aplastic anemia, hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia, erythropoiesis, osteoporosis, sarcoidosis, fibrosis of the lung, idiopathic interstitial pneumonia, dermatomyositis, common vitiligo, common ichthyosis, photoallergic sensitivity, cutaneous T-cell lymphoma, chronic lymphocytic leukemia, arteriosclerosis, Atherosclerosis, aortitis syndrome, polyarteritis nodosa, cardiomyopathy, scleroderma, Wegener's granulomatosis, Sjögren's syndrome, obesity, eosinophilic fasciitis, gingival lesions, periodontal ligament lesions, alveolar bone lesions, cementum lesions, glomerulonephritis, male pattern baldness or senile hair loss by preventing hair loss or providing hair generation and / or promoting hair generation and hair growth, muscular dystrophy, pyoderma and Sezary's syndrome, Addison's disease, ischemia-reperfusion injury after organ preservation, transplantation or ischemic disease, endotoxin shock, pseudomembranous colitis, colitis caused by drugs or radiation, ischemic acute renal insufficiency, chronic renal insufficiency, toxic diseases caused by pulmonary oxygen or drugs, lung cancer, emphysema, cataracts, siderosis,Retinitis pigmentosa, age-related macular degeneration, vitreous scars, corneal alkali burns, erythema polymorphum, linear IgA bullous dermatitis and cement dermatitis, gingivitis, periodontitis, sepsis, pancreatitis, diseases caused by environmental pollution, aging, carcinogenesis, metastasis and hypotension, diseases caused by histamine or leukotriene-C4 release, Behcet's disease, autoimmune hepatitis, primary biliary cirrhosis, sclerosing cholangitis, partial liver resection, acute liver necrosis, necrosis caused by toxins, viral hepatitis, shock or hypoxia, viral hepatitis B, non-A / non-B hepatitis, cirrhosis, alcoholic cirrhosis, liver failure, fulminant liver failure, late liver failure, "acute-on-chronic" liver failure, enhanced chemotherapy effects, cytomegalovirus infection, HCMV infection, AIDS, cancer, Alzheimer's disease, Parkinson's disease, trauma, CRS, ARDS, AKI or chronic bacterial infection. , 25. The method of claim 20, wherein the disease or condition comprises a lymphoid neoplasm selected from the group consisting of myeloproliferative neoplasms (MPNs) excluding polycythemia vera, myeloid / lymphoid neoplasms with PDGFRA rearrangement, myeloid / lymphoid neoplasms with PDGFRB rearrangement, myeloid / lymphoid neoplasms with FGFR1 rearrangement, myeloid / lymphoid neoplasms with PCM1-JAK2, myelodysplastic / myeloproliferative neoplasms (MDS / MPNs), myeloid sarcoma, myeloproliferation associated with Down syndrome, blastic plasmacytoid dendritic cell neoplasm, B lymphoblastic leukemia / lymphoma; and / or T lymphoblastic leukemia / lymphoma.

26. The method of claim 25, wherein the lymphoid neoplasm is a myeloproliferative neoplasm selected from the group consisting of chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), primary myelofibrosis (PMF), essential thrombocythaemia, chronic eosinophilic leukemia, or a combination thereof.

27. The method of claim 26, wherein the lymphoid tumor is chronic myeloid leukemia.

28. The method of claim 25, wherein the lymphoid tumor is a myelodysplastic / myeloproliferative neoplasm selected from the group consisting of chronic myelomonocytic leukemia, atypical chronic myeloid leukemia (aCML), juvenile myelomonocytic leukemia (JMML), MDS / MPN with ring sideroblasts and thrombocythemia (MDS / MPN-RS-T), or a combination thereof.

29. The method of claim 25, wherein the method further comprises identifying a subject suffering from the lymphoid tumor.

30. The method of claim 29, wherein the lymphoid neoplasm is chronic myelomonocytic leukemia, and identifying the subject comprises identifying a persistent peripheral blood monocytosis of ≥ 1×10 9 / L and monocytes accounting for ≥10% of the white blood cell (WBC) differential count and no observed rearrangements in the PDGFRA, PDGFRB, or FGFR1 genes and the PCM1-JAK2 fusion gene.

31. A compound according to any one of claims 1-16 or a composition according to claim 17 for use in a method of treatment.

32. A compound for use according to claim 31, wherein the method of treatment is a method for treating a disease or condition requiring an IRAK inhibitor.

33. A compound for use according to claim 31 or claim 32, wherein the disease or condition comprises an autoimmune disease, an inflammatory disorder, a cardiovascular disease, a neurodegenerative disorder, an allergic disorder, multiple organ failure, kidney disease, platelet aggregation, a hyperproliferative disorder, transplantation, sperm motility, red blood cell deficiency, transplant rejection, lung injury, respiratory disease, ischemic disease, bacterial infection, viral infection, immunomodulatory disorder, sickle cell disease, chemical or radiation induced lung injury, hemorrhagic fever or a combination thereof.

34. Use of a compound according to any one of claims 1 to 16 or a composition according to claim 17 in the manufacture of a medicament for in vivo treatment.

35. Use of a compound or composition according to claim 34, wherein the medicament is a medicament for treating a disease or condition requiring an IRAK inhibitor.

36. The use of a compound or composition according to claim 34 or claim 35, wherein the disease or disorder comprises an autoimmune disease, an inflammatory disorder, a cardiovascular disease, a neurodegenerative disorder, an allergic disorder, multiple organ failure, kidney disease, platelet aggregation, a hyperproliferative disorder, transplantation, sperm motility, red blood cell deficiency, transplant rejection, lung injury, respiratory disease, ischemic disease, bacterial infection, viral infection, immunomodulatory disorder, sickle cell disease, chemical or radiation induced lung injury, hemorrhagic fever, or a combination thereof.

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