Cyclopropanamide-containing compounds and uses thereof

Abstract

This application relates to the field of medicinal chemistry, specifically to cyclopropanamide-containing compounds represented by formula (I) and their uses, which have great medical value and market potential for autoimmune diseases and myeloproliferative neoplasms. [Formula 1] TIFF2025501662000047.tif33170

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to a Chinese patent application filed on January 18, 2022, bearing application number 202210052435.5 and entitled "Cyclopropanamide-containing compounds and their uses," the entire contents of which are incorporated herein by reference. [Technical Field]

[0002] This application relates to the field of medicinal chemistry, and specifically to cyclopropanamide-containing compounds and their uses. [Background technology]

[0003] Protein kinases are a family of enzymes that catalyze the phosphorylation of specific residues in proteins and are broadly classified as tyrosine and serine / threonine kinases. Inappropriate kinase activity due to mutation, overexpression or improper regulation, dysregulation or disregulation, and overproduction or underproduction of growth factors and cytokines is associated with numerous diseases, including, but not limited to, cancer, cardiovascular disease, hypersensitivity reactions, asthma and other respiratory diseases, autoimmune diseases, inflammatory diseases, bone diseases, metabolic disorders, and neurological and neurodegenerative diseases (e.g., Alzheimer's disease). Thus, protein kinases have become an important class of enzymes as target spots for therapeutic intervention.

[0004] The Janus kinase (JAK) family is a type of non-receptor protein tyrosine kinase that plays an important role in cytokine receptor signaling pathways through interactions with signal transducers and activators of transcription (STATs). STATs are a group of cytoplasmic proteins that can bind to target gene regulatory DNA. As downstream substrates of JAKs, STATs are activated by tyrosine phosphorylation under stimulation of external signals, and then enter the cell nucleus to regulate gene transcription.

[0005] The regulation of many aberrant immune responses, including autoimmune diseases such as allergy, asthma, (xeno)transplant rejection, rheumatoid arthritis, amyotrophic lateral sclerosis and multiple sclerosis, as well as hematological malignancies such as myeloproliferative disorders, leukemias and lymphomas, are all associated with the JAK / STAT signaling pathway.

[0006] The Janus kinase (JAK) family includes four members: JAK1, JAK2, JAK3, and TYK2. JAK1, JAK2, and TYK2 are widely distributed in various tissues and cells, while JAK3 is mainly distributed in lymphocytes.

[0007] JAK1 can bind to IL-10, IL-19, IL-20, IL-22, IL-26, IL-28, IFN-α, IFN-γ, IL-6 of the gp130 family, and other γc-containing receptors (Rodig S.J., et al., Cell, 1998, 93:373-383). JAK1 is a novel target spot in the field of immune-related diseases, inflammatory diseases, and cancer. JAK1 inhibitors can be used to treat / prevent autoimmune inflammatory diseases such as leukemia, lymphoma, melanoma, arthritis, psoriasis, lupus erythematosus, and acquired immunodeficiency syndrome (AIDS) (Hou S., et al., HμM. Genet., 2013, 132:1049-1058) (Hornakova T., et al., Blood, 2010, 115:3287-3295).

[0008] JAK2 plays an important role in the regulation of multiple receptor signals, including those of IL-3, IFN-γ, EPO, GH, etc. (Levy D.E., et al., Nat. Rev. Mol. Cell Biol., 2002, 3:651-662). JAK2 inhibitors have great medical value and market potential for the treatment of diseases such as myelofibrosis, solid tumors, and hematological tumors.

[0009] JAK3 regulates cell signaling by binding to the common gamma chain (γc) in the receptor complexes for cytokines such as IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21. JAK3 inhibitors can block T cell activation and prevent transplant rejection after transplant surgery. Furthermore, JAK3 inhibitors can also regulate the functions of lymphocytes, macrophages, and mast cells. JAK3 inhibitors are expected to be used for the treatment or prevention of several diseases associated with the function of lymphocytes, macrophages, or mast cells.

[0010] Torfatinib is the first FDA-approved novel oral JAK inhibitor, acting on JAK1 and JAK3, and is used to treat rheumatoid arthritis (RA). However, while torfatinib alleviates RA symptoms, it also causes some side effects, including the development of certain infections, malignancies, and lymphomas. Studies have shown that the overall risk of infection and mortality associated with torfatinib is similar to that of treating RA with biologics. Given the pleiotropic effects of JAKs on many regulatory pathways and immune processes, non-selective JAK inhibitors pose a risk of adverse reactions, and selective JAK inhibitors have become an important area of ​​current research.

[0011] Filgotinib, a new-generation JAK1 selective inhibitor developed by Galapagos, Belgium, reduces the risk of anemia or infection associated with tolfatinib. However, filgotinib has relatively weak activity, with an IC50 of greater than 10 nM for JAK1, and its clinical dose is relatively high (Expert Opin. Investig. Drugs. 2016, 25, 1355).

[0012] Although a series of JAK inhibitors have been disclosed on the market, there is still a need to develop new compounds with better efficacy and JAK selectivity. Through continuous efforts, the present application has developed a compound having a structure represented by general formula (I), and found that the compound having such a structure exhibits excellent efficacy. Summary of the Invention [Problem to be solved by the invention]

[0013] The present application provides a cyclopropanamide-containing compound selected from compounds represented by general formula (I), their stereoisomers, and pharmaceutically acceptable salts thereof. The stereoisomers include enantiomers, diastereomers, meso compounds, and racemates. [Means for solving the problem]

[0014] To achieve the above objectives, the present application employs the following technical solutions:

[0015] The present application provides a compound represented by general formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. [ka] During the ceremony, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; TIFF0007784561000002.tif22170 where, m is an integer from 1 to 4; TIFF0007784561000003.tif48170R is selected from hydrogen, a C1 to C6 alkyl group, a C1 to C6 alkoxy group, or a halogen; Z1, Z2, and Z3 are each independently selected from —C(R3R4)—, —S(O)2—, —S(O)—, —CH(S(O)2R′)—, or —CH(S(O)2NR′R″)—; R1 and R2 are each independently selected from hydrogen, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C7 cycloalkyl group; R' and R'' are each independently selected from a substituted or unsubstituted C1-C6 alkyl group or a C3-C7 cycloalkyl group, and the substituted group is selected from the group consisting of halogen, a C1-C6 alkoxy group, a C1-C6 alkylthio group, a C3-C7 cycloalkyl group, a hydroxy group, an amino group, an NHC1-C6 alkyl group, and an N(C1-C6 alkyl)2 group; R3 and R4 are each independently selected from hydrogen, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C7 cycloalkyl group.

[0016] Preferably, the above compound, its stereoisomer or its pharmaceutically acceptable salt has a structure represented by general formula (II). [ka] During the ceremony, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; m is an integer from 1 to 4; TIFF0007784561000005.tif22170R is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkoxy group, or a halogen; Z1, Z2 and Z3 are each independently selected from -C(R3R4)-, -S(O)2-, -S(O)-, -CH(S(O)2R')-, or -CH(S(O)2NR'R'')-, and when Y is hydrogen, Z2 is not -S(O)2-; R' and R'' are each independently selected from a substituted or unsubstituted C1-C6 alkyl group or a C3-C7 cycloalkyl group, and the substituted group is selected from the group consisting of halogen, a C1-C6 alkoxy group, a C1-C6 alkylthio group, a C3-C7 cycloalkyl group, a hydroxy group, an amino group, an NHC1-C6 alkyl group, and an N(C1-C6 alkyl)2 group; R3 and R4 are each independently selected from hydrogen, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C7 cycloalkyl group.

[0017] Preferably, the above compound, its stereoisomer or its pharmaceutically acceptable salt has a structure represented by general formula (III). [ka] During the ceremony, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; TIFF0007784561000007.tif22170 R1 and R2 are each independently selected from hydrogen, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C7 cycloalkyl group; R' is selected from a substituted or unsubstituted C1-C6 alkyl group or a C3-C7 cycloalkyl group, and the substituted group is selected from the group consisting of halogen, a C1-C6 alkoxy group, a C1-C6 alkylthio group, a C3-C7 cycloalkyl group, a hydroxy group, an amino group, an NHC1-C6 alkyl group, and an N(C1-C6 alkyl group)2.

[0018] Preferably, the above compound, its stereoisomer or its pharmaceutically acceptable salt has a structure represented by general formula (IV). [ka] During the ceremony, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; m is an integer from 1 to 4; R is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkoxy group, or a halogen; Z1 is selected from -C(R3R4)-, -S(O)2-, -S(O)-, -CH(S(O)2R')-, or -CH(S(O)2NR'R'')-; R' and R'' are each independently selected from a substituted or unsubstituted C1-C6 alkyl group or a C3-C7 cycloalkyl group, and the substituted group is selected from the group consisting of halogen, a C1-C6 alkoxy group, a C1-C6 alkylthio group, a C3-C7 cycloalkyl group, a hydroxy group, an amino group, an NHC1-C6 alkyl group, and an N(C1-C6 alkyl)2 group; R3 and R4 are each independently selected from hydrogen, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C7 cycloalkyl group.

[0019] Preferably, the above compound, its stereoisomer or its pharmaceutically acceptable salt has a structure represented by general formula (V). [ka] During the ceremony, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; m is an integer from 1 to 4; R is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkoxy group, or a halogen; Z2 is selected from -C(R3R4)-, -S(O)2-, -S(O)-, -CH(S(O)2R')-, or -CH(S(O)2NR'R'')-, and when Y is hydrogen, Z2 is not -S(O)2-; R' and R'' are each independently selected from a substituted or unsubstituted C1-C6 alkyl group or a C3-C7 cycloalkyl group, and the substituted group is selected from the group consisting of halogen, a C1-C6 alkoxy group, a C1-C6 alkylthio group, a C3-C7 cycloalkyl group, a hydroxy group, an amino group, an NHC1-C6 alkyl group, and an N(C1-C6 alkyl)2 group; R3 and R4 are each independently selected from hydrogen, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C7 cycloalkyl group.

[0020] Preferably, the above compound, its stereoisomer or its pharmaceutically acceptable salt has a structure represented by general formula (VI). [ka] During the ceremony, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; m is an integer from 1 to 4; R is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkoxy group, or a halogen; Z3 is selected from -C(R3R4)-, -S(O)2-, -S(O)-, -CH(S(O)2R')-, or -CH(S(O)2NR'R'')-; R' and R'' are each independently selected from a substituted or unsubstituted C1-C6 alkyl group or a C3-C7 cycloalkyl group, and the substituted group is selected from the group consisting of halogen, a C1-C6 alkoxy group, a C1-C6 alkylthio group, a C3-C7 cycloalkyl group, a hydroxy group, an amino group, an NHC1-C6 alkyl group, and an N(C1-C6 alkyl)2 group; R3 and R4 are each independently selected from hydrogen, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C7 cycloalkyl group.

[0021] Preferably, the above compound, its stereoisomer or its pharmaceutically acceptable salt has a structure represented by general formula (VII). [ka] During the ceremony, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; R1 and R2 are each independently selected from hydrogen, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C7 cycloalkyl group.

[0022] Preferably, the above compound, its stereoisomer or its pharmaceutically acceptable salt has a structure represented by general formula (VIII). [ka] During the ceremony, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; R' is selected from a substituted or unsubstituted C1-C6 alkyl group or a C3-C7 cycloalkyl group, and the substituted group is selected from the group consisting of halogen, a C1-C6 alkoxy group, a C1-C6 alkylthio group, a C3-C7 cycloalkyl group, a hydroxy group, an amino group, an NHC1-C6 alkyl group, and an N(C1-C6 alkyl group)2.

[0023] Preferably, Y is selected from H or F.

[0024] Preferably, in the above general formula Selected from TIFF0007784561000013.tif15170.

[0025] Preferably, in the above general formula (IV), Z1 is selected from -CH(SO2R')-, where R' is selected from an unsubstituted C1-C6 alkyl group or a C3-C7 cycloalkyl group, and R is selected from hydrogen, a methyl group, a methoxy group, or a halogen.

[0026] Preferably, in the above general formula (V), when Y is hydrogen, Z2 is selected from -CR3R4-, or -CH(S(O)2R')-, further wherein R3 and R4 are each independently selected from hydrogen, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, or a C3-C7 cycloalkyl group; R' is selected from a C1-C6 alkyl group or a C3-C7 cycloalkyl group; R is selected from hydrogen, a methyl group, a methoxy group, or a halogen; and when Y is halogen, Z2 is selected from -SO2-, and R is selected from hydrogen, a methyl group, a methoxy group, or a halogen.

[0027] Preferably, in the above general formula (VI), Z3 is selected from -SO2- or -S(O)-, and R is selected from hydrogen, a methyl group, a methoxy group, or a halogen.

[0028] Preferably, in the above general formula (VII), R1 and R2 are each independently selected from hydrogen, fluorine, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a t-butyl group, an s-butyl group, an n-pentyl group, a trifluoromethyl group, a cyclopropyl group, and a cyclopentyl group.

[0029] Preferably, in the above general formula (VIII), R' is selected from a substituted or unsubstituted methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, t-butyl group, s-butyl group, n-pentyl group, isopentyl group, cyclopropyl group, and cyclopentyl group, and the substituted group is selected from the group consisting of halogen, C1-C6 alkoxy group, C1-C6 alkylthio group, C3-C7 cycloalkyl group, hydroxy group, amino group, NHC1-C6 alkyl group, and N(C1-C6 alkyl group)2.

[0030] Furthermore, the present application further provides a compound represented by the following formula, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: TIFF0007784561000014.tif137170

[0031] Furthermore, the present application further provides a compound represented by the following formula, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: TIFF0007784561000015.tif202170

[0032] Additionally, the present application further provides a pharmaceutical composition comprising a therapeutically effective amount of the above compound, its stereoisomer or a pharmaceutically acceptable salt component thereof, and a pharmaceutically acceptable carrier.

[0033] Furthermore, the present application further provides use of the above-mentioned compound, its stereoisomer or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical composition in the manufacture of a medicament for treating a disease associated with an abnormality in the JAK signaling pathway.

[0034] Preferably, the disease is an autoimmune disease.

[0035] More preferably, the autoimmune disease is selected from rheumatoid arthritis, ulcerative colitis, atopic dermatitis, or systemic lupus erythematosus.

[0036] Preferably, the disease is a myeloproliferative neoplasm.

[0037] More preferably, the myeloproliferative neoplasm is selected from essential thrombocytosis, polycythemia vera, or primary myelofibrosis.

[0038] Furthermore, the present application further provides a method for treating a disease associated with an abnormality in the JAK signaling pathway, the method comprising administering to a subject in need thereof a therapeutically effective amount of the above-mentioned compound, its stereoisomer or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical composition.

[0039] Preferably, the disease is an autoimmune disease.

[0040] More preferably, the autoimmune disease is selected from rheumatoid arthritis, ulcerative colitis, atopic dermatitis, or systemic lupus erythematosus.

[0041] Preferably, the disease is a myeloproliferative neoplasm.

[0042] More preferably, the myeloproliferative neoplasm is selected from essential thrombocytosis, polycythemia vera, or primary myelofibrosis.

[0043] The "therapeutically effective amount" varies depending on the subject, target organ, symptoms, administration method, etc., and can be determined by the judgment of a physician, taking into consideration the type of dosage form, administration method, age and weight of the patient, symptoms of the patient, etc. [Effects of the Invention]

[0044] 1) The compounds of the present application have good inhibitory activity against JAK1, JAK2 or JAK3 kinase, and have clear selectivity against JAK1, JAK2 or JAK3 kinase.

[0045] 2) The compounds of the present application have obvious pharmacokinetic advantages, provide more options for the prevention and / or treatment of diseases associated with abnormalities in the JAK signaling pathway, and have good prospects for clinical application. DETAILED DESCRIPTION OF THE INVENTION

[0046] In order to clarify the objectives, technical solutions and advantages of the embodiments of the present application, the technical solutions in the embodiments of the present application will be described below clearly and completely, but it is obvious that the described embodiments are only some of the embodiments of the present application, and not all of the embodiments. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without creative efforts fall within the scope of protection of the present application.

[0047] Unless otherwise stated, terms used in the specification and claims have the following meanings:

[0048] The term "alkyl group" refers to a saturated aliphatic hydrocarbon group, including straight and branched chain groups of 1 to 20 carbon atoms. Preferably, the alkyl group contains 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, most preferably 1 to 4 carbon atoms, and most preferably methyl.

[0049] Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, s-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, and 5-methylhexyl. , 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, and non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, s-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like.The alkyl group may be substituted or unsubstituted, and when substituted, the substitution group may be at any available attachment point, and is preferably one or more groups independently selected from alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, alkylthio groups, alkylamino groups, halogens, sulfhydryl groups, hydroxy groups, nitro groups, cyano groups, cycloalkyl groups, heterocyclic alkyl groups, aryl groups, heteroaryl groups, cycloalkoxy groups, heterocyclic alkoxy groups, cycloalkylthio groups, heterocyclic alkylthio groups, oxo groups, amino groups, haloalkyl groups, hydroxyalkyl groups, carboxyl groups, and carboxylic acid ester groups.

[0050] The term "cycloalkyl group" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon-substituted group containing 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 7 carbon atoms, and is optimally a cyclopropyl group or a cyclopentyl group. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentene, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene (cycloheptatrienyl), cyclooctyl, etc., preferably cyclopropyl or cyclopentyl. Polycyclic cycloalkyl groups include spirocyclic, fused, and bridged cycloalkyl groups. The cycloalkyl group may be optionally substituted or unsubstituted, and when substituted, the substituted groups are one or more groups independently selected from alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, alkylthio groups, alkylamino groups, halogens, sulfhydryl groups, hydroxy groups, nitro groups, cyano groups, cycloalkyl groups, heterocyclic alkyl groups, aryl groups, heteroaryl groups, cycloalkoxy groups, heterocyclic alkoxy groups, cycloalkylthio groups, heterocyclic alkylthio groups, oxo groups, amino groups, haloalkyl groups, hydroxyalkyl groups, carboxyl groups, and carboxylic acid ester groups.

[0051] The term "alkoxy group" refers to -O-(alkyl group) and -O-(unsubstituted cycloalkyl group), where alkyl group and cycloalkyl group are defined above. Non-limiting examples include methoxy group, ethoxy group, propoxy group, butoxy group, cyclopropoxy group, cyclobutoxy group, cyclopentyloxy group, cyclohexyloxy group, etc. An alkoxy group may be optionally substituted or unsubstituted. When substituted, the substituted groups are one or more groups independently selected from alkyl group, alkenyl group, alkynyl group, alkoxy group, alkylthio group, alkylamino group, halogen, sulfhydryl group, hydroxy group, nitro group, cyano group, cycloalkyl group, heterocycloalkyl group, aryl group, heteroaryl group, cycloalkoxy group, heterocycloalkoxy group, cycloalkylthio group, heterocycloalkylthio group, amino group, haloalkyl group, hydroxyalkyl group, carboxyl group, and carboxylic acid ester group.

[0052] A "haloalkyl group" refers to an alkyl group substituted with one or more halogens, where alkyl is defined above.

[0053] "Hydroxy" refers to an --OH group.

[0054] A "hydroxyalkyl group" refers to an alkyl group substituted with a hydroxy group, where alkyl is as defined above.

[0055] "Halogen" refers to fluorine, chlorine, bromine or iodine, preferably fluorine or iodine.

[0056] An "amino group" refers to -NH2.

[0057] A "cyano group" refers to -CN.

[0058] A "nitro group" refers to -NO2.

[0059] An "oxo group" refers to =O.

[0060] A "carboxyl group" refers to -C(O)OH.

[0061] The term "carboxylic acid ester group" refers to -C(O)O (alkyl group) or (cycloalkyl group), where the alkyl group and cycloalkyl group are defined as above.

[0062] "Optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "an optional heterocycloalkyl group substituted with an alkyl group" means that the alkyl group may be present but is not required to be present, and the description includes situations where the heterocycloalkyl group is substituted with an alkyl group and situations where the heterocycloalkyl group is not substituted with an alkyl group.

[0063] "Substituted" means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, are independently replaced with a corresponding number of substitution groups. Needless to say, substitution groups are present only at their possible chemical positions, and a person skilled in the art can determine possible or impossible substitutions (by experiment or theory) without much effort. For example, an amino group or a hydroxy group having free hydrogen may be unstable when bonded to a carbon atom having an unsaturated (e.g., alkene) bond.

[0064] The technical solution of the present application will be further described below with reference to specific examples. Unless otherwise specified, the instruments, consumables, reagents, etc. used in the following examples can all be obtained through conventional commercial means, and the experimental methods for which no specific conditions are specified in the examples can be selected according to conventional methods and conditions or product instructions.

[0065] Example 1: Synthesis of Compounds 1 to 11 TIFF0007784561000016.tif142170Synthetic route of general formula compound (IX) to which compounds 1-11 belong Step 1: Diethyl cyanomethylphosphonate (177 mg, 1 mmol) was dissolved in dry tetrahydrofuran (10 mL). Sodium hydride (60 mg, 1.5 mmol) was added in an ice bath. After stirring for 1 hour, the corresponding aldehyde (1 mmol) was added. The reaction was stirred overnight at room temperature. After completion of the reaction, the mixture was quenched by adding saturated ammonium chloride solution (20 mL). The mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was used directly in the next step. Step 2: N-(5-(1H-pyrazol-4-yl)imidazole[1,2-a]pyridin-2-yl)cyclopropanecarboxamide (134 mg, 0.5 mmol), the residue from the previous step (0.6 mmol), and 1,8-diazabicyclo[5,4,0]undecene-7 (2 drops) were dissolved in acetonitrile and heated to 80 °C for 8 hours. After returning to room temperature, water (20 mL) was added to the reaction mixture, which was then concentrated to remove most of the acetonitrile. The mixture was then extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to give the compound.

[0066] Example 2: Synthesis of Compound 1 Following the synthesis route of Example 1, compound 1 (70 mg) was obtained using cyclopentylformaldehyde (cyclopentanecarbaldehyde) in step 1, with a yield of 36%. LCMS (ESI+): 389.42 (M+H). + .

[0067] Example 3: Synthesis of Compound 2 Following the synthesis route of Example 1, compound 2 (100 mg) was obtained using cyclopropylformaldehyde (cyclopropanecarboxaldehyde) in step 1, with a yield of 56%. LCMS (ESI+): 361.42 (M+H). + .

[0068] Example 4: Synthesis of Compound 3 Following the synthetic route of Example 1, compound 3 (73 mg) was obtained using 2-cyclopropylacetaldehyde in step 1, with a yield of 39%. LCMS (ESI+): 375.38 (M+H). + .

[0069] Example 5: Synthesis of Compound 4 Following the synthetic route of Example 1, compound 4 (80 mg) was obtained using propionaldehyde in step 1 in 41% yield. LCMS (ESI+): 349.16 (M+H). + .

[0070] Example 6: Synthesis of Compound 5 Following the synthetic route of Example 1, butyraldehyde was used in step 1 to obtain compound 5 (79 mg) in 43% yield. LCMS (ESI+): 363.20 (M+H). + .

[0071] Example 7: Synthesis of Compound 6 Following the synthetic route of Example 1, compound 6 (55 mg) was obtained using 2-methoxyacetaldehyde in step 1, with a yield of 30%. LCMS (ESI+): 365.06 (M+H). + .

[0072] Example 8: Synthesis of Compound 7 Following the synthetic route of Example 1, compound 7 (72 mg) was obtained using valeraldehyde in step 1, with a yield of 38%. LCMS (ESI+): 377.08 (M+H). + .

[0073] Example 9: Synthesis of Compound 8 Following the synthetic route of Example 1, compound 8 (87 mg) was obtained using 4-methylvaleraldehyde in step 1, with a yield of 46%. LCMS (ESI+): 377.12 (M+H). + .

[0074] Example 10: Synthesis of Compound 9 Following the synthetic route of Example 1, compound 9 (80 mg) was obtained using 3-methoxypropionaldehyde in step 1, with a yield of 42%. LCMS (ESI+): 379.09 (M+H). + .

[0075] Example 11: Synthesis of Compound 10 Following the synthetic route of Example 1, compound 10 (105 mg) was obtained using 4,4,4-trifluorobutyraldehyde in step 1, with a yield of 50%. LCMS (ESI+): 417.16 (M+H). + .

[0076] Example 12: Synthesis of Compound 11 The rest of the synthesis was carried out according to the synthetic route of Example 1, except that N-(5-(1H-pyrazol-4-yl)imidazole[1,2-a]pyridin-2-yl)cyclopropanecarboxamide was used instead of N-(5-(1H-pyrazol-4-yl)imidazole[1,2-a]pyridin-2-yl)cyclopropanecarboxamide in Step 2, and cyclopentylformaldehyde was used in Step 1 to obtain compound 11 (79 mg), with a yield of 41%. LCMS (ESI+): 390.25 (M+H). + .

[0077] Example 13: Synthesis of Compounds 12, 13, and 14 TIFF0007784561000017.tif73170Synthetic route for compound (X) of the general formula to which compounds 12, 13, and 14 belong Step 1: Diethyl cyanomethylphosphonate (177 mg, 1 mmol) was dissolved in dry tetrahydrofuran (10 mL). Sodium hydride (60 mg, 1.5 mmol) was added in an ice bath. After stirring for 1 hour, the cyclohexanone derivative (1 mmol) was added. The reaction was stirred overnight at room temperature. The mixture was quenched by adding saturated ammonium chloride solution (20 mL) and extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was used directly in the next step. Step 2: N-(5-(1H-pyrazol-4-yl)imidazole[1,2-a]pyridin-2-yl)cyclopropanecarboxamide (135 mg, 0.5 mmol), the residue from Step 1 (0.6 mmol), and 1,8-diazabicyclo[5,4,0]undecene-7 (2 drops) were dissolved in acetonitrile and heated to 80 °C for 8 hours. After returning to room temperature, water (20 mL) was added to the reaction mixture, which was then concentrated to remove most of the acetonitrile. The mixture was then extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to give the compound.

[0078] Example 14: Synthesis of Compound 12 Following the synthetic route of Example 13, compound 12 (74 mg) was obtained using 4,4-difluorocyclohexanone in step 1, with a yield of 35%. LCMS (ESI+): 425.02 (M+H). + .

[0079] Example 15: Synthesis of Compound 13 Following the synthetic route of Example 13, compound 13 (113 mg) was obtained using 4-trifluoromethylcyclohexanone in step 1, with a yield of 49%. LCMS (ESI+): 457.01 (M+H). + .

[0080] Example 16: Synthesis of Compound 14 Following the synthetic route of Example 13, compound 14 (99 mg) was obtained using cyclohexanone in step 1 in 50% yield. LCMS (ESI+): 389.08 (M+H). + .

[0081] Example 17: Synthesis of Compound 15 TIFF0007784561000018.tif34170 Step 1: (4-(Bromomethyl)phenyl)-boronic acid (215 mg, 1 mmol) and 3-(methylsulfone)azetidine (135 mg, 1 mmol) were dissolved in acetonitrile, and then potassium carbonate (207 mg, 1 mmol) was added in batches and the reaction was carried out at room temperature overnight. The reaction was filtered, and the filtrate was concentrated. The resulting residue was used directly in the next step. Step 2: The residue from the previous step (135 mg, 0.5 mmol), N-(5-bromoimidazo[1,2-a]pyridin-2-yl)cyclopropanecarboxamide (140 mg, 1 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (36 mg, 0.1 mmol), and potassium carbonate (138 mg, 2 mmol) were dissolved in dioxane (10 mL) and water (10 mL). The mixture was purged with nitrogen three times and stirred at 110 °C for 8 h. After returning to room temperature, water (20 mL) was added to the reaction mixture, which was then extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane:methanol=100:0-100:5) to give compound 15 (119 mg) in a 56% yield. LCMS (ESI+): 212.99 (M / 2+H). + .

[0082] Example 18: Synthesis of Compound 16 TIFF0007784561000019.tif36170 Step 1: (4-(Bromomethyl)phenyl)-boronic acid (108 mg, 0.5 mmol) and 4-(methylsulfone)piperidine (84 mg, 0.5 mmol) were dissolved in acetonitrile (5 mL), and then potassium carbonate (138 mg, 1 mmol) was added in batches. The reaction was allowed to proceed at room temperature overnight, filtered, and the filtrate was concentrated. The resulting residue was used directly in the next step. Step 2: The residue obtained in step 1 was dissolved in dioxane (10 mL) and water (10 mL). N-(5-bromoimidazo[1,2-a]pyridin-2-yl)cyclopropanecarboxamide (140 mg, 0.5 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (36 mg, 0.1 mmol), and potassium carbonate (138 mg, 2 mmol) were added. The mixture was purged with nitrogen three times and stirred at 110 °C for 8 hours. After returning to room temperature, water (20 mL) was added to the reaction mixture, which was then extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to give compound 16 (108 mg) in 48% yield. LCMS(ESI+):227.13(M / 2+H) + .

[0083] Example 19: Synthesis of Compound 17 TIFF0007784561000020.tif86170 Step 1: Tetrahydro-2H-thiopyran-4-ol-1,1-dioxide (150 mg, 1 mmol) and p-toluenesulfonyl chloride (215 mg, 1 mmol) were dissolved in pyridine (5 mL) and reacted overnight at room temperature. After concentration, the residue was purified by column chromatography (petroleum ether:ethyl acetate = 100:0 to 50:50) to obtain the compound methyl 1,1-dioxotetrahydro-2H-thiopyran-4-yl-4-benzenesulfonate. Step 2: p-Hydroxybenzeneboronic acid (138 mg, 1 mmol), N-(5-bromoimidazo[1,2-a]pyridin-2-yl)cyclopropanecarboxamide (279 mg, 1 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (73 mg, 0.1 mmol), and potassium carbonate (276 mg, 2 mmol) were dissolved in dioxane (10 mL) and water (10 mL). The mixture was purged with nitrogen three times and stirred at 100 °C for 8 hours. After returning to room temperature, water (20 mL) was added to the reaction mixture, which was then extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by column chromatography (dichloromethane:methanol=100:0 to 100:5) to obtain the compound N-(5-(4-hydroxyphenyl)imidazole[1,2-a]pyridin-2-yl)cyclopropanecarboxamide. Step 3: Methyl 1,1-dioxotetrahydro-2H-thiopyran-4-yl-4-benzenesulfonate (152 mg, 0.5 mmol) obtained in Step 1, N-(5-(4-hydroxyphenyl)imidazole[1,2-a]pyridin-2-yl)cyclopropanecarboxamide (146 mg, 0.5 mmol) obtained in Step 2, and potassium carbonate (138 mg, 1 mmol) were added to N,N-dimethylformamide (5 mL) and heated to 80 °C for 5 h. After returning to room temperature, water (20 mL) was added to the reaction mixture, which was then extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to give compound 17 (137 mg) in 62% yield. LCMS(ESI+):425.98(M+H) + .

[0084] Example 20: Synthesis of Compound 18 Following the synthetic route of Example 19, compound 18 (104 mg) was obtained in 50% yield by using 2-fluoro-4-hydroxyphenylboronic acid instead of p-hydroxybenzeneboronic acid in step 2. LCMS (ESI+): 443.98 (M+H). + .

[0085] Example 21: Synthesis of Compound 19 Following the synthetic route of Example 19, N-(5-bromo-[1,2,4]triazole[1,5-a]pyridin-2-yl)cyclopropanecarboxamide was used in place of N-(5-bromoimidazo[1,2-a]pyridin-2-yl)cyclopropanecarboxamide in Step 2 to obtain compound 19 (58 mg) in 43% yield. LCMS (ESI+): 427.15 (M+H). + .

[0086] Example 22: Synthesis of Compound 20 TIFF0007784561000023.tif40170 Step 1: (2-Fluoro-4-formylphenyl)boronic acid (168 mg, 1 mmol), 4-(methylsulfone)piperidine (135 mg, 1 mmol), and one drop of acetic acid were added to methanol and stirred at room temperature for 2 hours. Sodium cyanoborohydride (126 mg, 2 mmol) was then added and stirring was continued overnight. The reaction mixture was concentrated, followed by the addition of water (20 mL) and extraction with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was used directly in the next step. Step 2: The residue from Step 1 was dissolved directly in dioxane (10 mL) and water (10 mL). N-(5-bromoimidazo[1,2-a]pyridin-2-yl)cyclopropanecarboxamide (140 mg, 0.5 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (36 mg, 0.05 mmol), and potassium carbonate (138 mg, 1 mmol) were added. The mixture was purged with nitrogen three times and stirred at 100 °C for 8 h. After returning to room temperature, water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to give compound 20 (120 mg) in 51% yield. LCMS(ESI+):236.13(M / 2+H) + .

[0087] Example 23: Synthesis of Compound 21 TIFF0007784561000024.tif73170 Step 1: Tetrahydrothiopyran-4-ol (590 mg, 5 mmol) and p-toluenesulfonyl chloride (1.1 g, 6 mmol) were dissolved in pyridine (10 mL) and reacted overnight at room temperature. After concentration, the residue was purified by column chromatography (petroleum ether:ethyl acetate = 100:0 to 50:50) to obtain the compound tetrahydro-2H-thiopyran-4-yl-4-benzenesulfonate. Step 2: p-Hydroxybenzeneboronic acid (1.38 g, 10 mmol), N-(5-bromo-[1,2,4]triazole[1,5-a]pyridin-2-yl)cyclopropanecarboxamide (2.81 g, 10 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (730 mg, 1 mmol), and potassium carbonate (2.76 g, 20 mmol) were dissolved in dioxane (20 mL) and water (20 mL). The mixture was purged with nitrogen three times and stirred at 110 °C for 8 h. After returning to room temperature, water (20 mL) was added to the reaction mixture, which was then extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane:methanol=100:0-100:5) to obtain the compound N-(5-(4-hydroxyphenyl)-[1,2,4]triazole[1,5-a]pyridin-2-yl)cyclopropanecarboxamide. Step 3: Methyl tetrahydro-2H-thiopyran-4-yl-4-benzenesulfonate (272 mg, 1 mmol) obtained in Step 1, N-(5-(4-hydroxyphenyl)-[1,2,4]triazole[1,5-a]pyridin-2-yl)cyclopropanecarboxamide (294 mg, 1 mmol) obtained in Step 2, and potassium carbonate (276 mg, 2 mmol) were added to N,N-dimethylformamide (5 mL), heated to 80 °C, and reacted for 5 hours. After returning to room temperature, water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane:methanol=100:0-100:5) to obtain the compound N-(5-(4-((1-tetrahydrothiopyran-4-yl)oxy)phenyl)-[1,2,4]triazole[1,5-a]pyridin-2-yl)cyclopropanecarboxamide. N-(5-(4-((1-tetrahydrothiopyran-4-yl)oxy)phenyl)-[1,2,4]triazole[1,5-a]pyridin-2-yl)cyclopropanecarboxamide (197 mg, 1 mmol) was dissolved in methanol (5 mL). N-bromosuccinimide (178 mg, 2 mmol) and potassium tert-butoxide (112 mg, 2 mmol) were added under ice-bath conditions, and the mixture was allowed to react overnight at room temperature. Saturated saline (20 mL) was added, and the mixture was extracted with ethyl acetate (10 mL × 3). The combined organic phase was washed with saturated saline (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to give compound 21 (66 mg) in a 32% yield. LCMS (ESI+): 411.03 (M+H). + .

[0088] Example 24: Synthesis of Compound 22 Step 1: t-Butyl (5-bromoimidazo[1,2-a]pyridin-2-yl)carbamate (311 mg, 1 mmol) was dissolved in dichloromethane (10 mL), trifluoroacetic acid (5 mL) was added in an ice bath, and the mixture was allowed to react at room temperature for 5 hours. The mixture was then concentrated. The resulting residue was used directly in the next step. Step 2: The residue from Step 1 was dissolved in dichloromethane (10 mL), trans-2-fluorocyclopropanecarboxylic acid (114 mg, 1.1 mmol) was added, and N,N-diisopropylethylamine (260 mg, 2 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (570 mg, 1.5 mmol) were added in an ice bath. The mixture was then reacted overnight at room temperature. After the reaction was completed, water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10 mL × 3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane:methanol=100:0-100:5) to obtain the compound trans-N-(5-bromoimidazo[1,2-a]pyridin-2-yl)-2-fluorocyclopropanecarboxamide. Step 3: trans-N-(5-bromoimidazo[1,2-a]pyridin-2-yl)-2-fluorocyclopropanecarboxamide (149 mg, 0.5 mmol), 4-{[4-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl}thiomorpholine-1,1-dione (175 mg, 0.5 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (36 mg, 0.05 mmol), and potassium carbonate (138 mg, 1 mmol) were dissolved in dioxane (10 mL) and water (10 mL). The mixture was purged with nitrogen three times and reacted at 110° C. with stirring for 8 hours. After returning to room temperature, water (20 mL) was added to the reaction mixture, which was then extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to give compound 22 (124 mg) in 56% yield. LCMS (ESI+): 443.55 (M+H). + . The resulting compound 22 was then resolved under the following resolution conditions: Compound 22 was first dissolved in dichloromethane (concentration 10 mg / mL), then diluted with ethanol to 2.5 mg / mL, and then resolved using an HPLC chiral column UniChiral CMD-5H (21.2 mm 1.D x 250 mmL) with ethanol / diethylamine (100:1) as the mobile phase at a flow rate of 12 mL / min, UV at 254 nm, and a column temperature of 25°C to obtain compounds 22-1 and 22-2, respectively. Retention times: Compound 22-1 was 10.69 min (>99% ee), and Compound 22-2 was 13.54 min (>99% ee).

[0089] Example 25: Synthesis of Compound 23 Following the synthetic route of Example 24, t-butyl (5-bromo-[1,2,4]triazole[1,5-a]pyridin-2-yl)carbamate was used in place of t-butyl (5-bromoimidazo[1,2-a]pyridin-2-yl)carbamate in Step 1 to obtain compound 23 (100 mg) in 45% yield. LCMS (ESI+): 444.17 (M+H). + . The resulting compound 23 was then resolved under the following conditions: Compound 23 was dissolved in ethanol (4 mg / mL) and purified using a HPLC chiral column UniChiral CMD-5H (21.2 mm 1.D x 250 mmL) with ethanol as the mobile phase, a flow rate of 12 mL / min, UV at 254 nm, and a column temperature of 25°C. Compounds 23-1 and 23-2 were obtained after resolution. Retention times: Compound 23-1 was 11.58 min (>99% ee), and Compound 23-2 was 16.47 min (>99% ee).

[0090] Example 26: Synthesis of Compounds 24-33 Compounds 24 to 33 were synthesized using appropriate raw materials and following the synthesis routes in Examples 24 and 25.

[0091] Specifically, the compound numbers, structures, synthesis methods and characteristic data are as follows: TIFF0007784561000027.tif245170TIFF0007784561000028.tif107170

[0092] Example 27: JAK1 / JAK2 / JAK3 kinase inhibitory activity test 1. Experimental consumables JAK1:Thermo Fisher PV4774 2059141D JAK2:Carna 08-045 14CBS-0374 H JAK3:Carna 08-046 19CBS-0798 B ATP (10 mM): CST 9804 DTT: 100 mM MgCl2: 1M MnCl2: 1M HTRF Kinase-TK kit (HTRF tyrosine kinase kit): cis-bio 62TK0PEC HTRF 96 well low volume plate (HTRF 96 well microplate): cis-bio 66PL96100

[0093] 2. Test conditions JAK1:0.32ng / μL, ATP 4μM, substrate 1μM, time 4h JAK2:0.008ng / μL, ATP 4μM, substrate 1μM, time 2h JAK3: 0.1ng / μL, ATP 3μM, substrate 1μM, time 3h

[0094] 3.JAK1 kinase inhibitory activity test 3.1 Preparation of reagents 1) Preparation of 1x kinase buffer: 5x kinase buffer was diluted with sterile water to 1x kinase buffer, and then 5mM MgCl2, 1mM MnCl2 and 1mM DTT were added. 2) Preparation of 5xJAK1: The concentration of JAK1 mother solution is 160 ng / μL, and the concentration is adjusted to 5x the final concentration, i.e., 1.6 ng / μL, and the mother solution is diluted 100 times. 3) Preparation of 5x ATP: 4 µM ATP was prepared at 5x its concentration, i.e., 20 µM, and directly diluted 500-fold from 10 mM ATP to obtain the required ATP concentration. 4) Preparation of 5x substrate: 5x of 1µM, i.e. 5µM, the substrate concentration was 500µM, diluted to 100x, i.e. 5µM substrate. 5) Preparation of 2.5x target compound: The concentration of the compound buffer was 10mM, and the treatment concentration started from 10μM. First, prepare 100x buffer, i.e., 1mM, and then dilute from 10mM to 10x. Then, dilute to a total of 10 concentrations with a 1:3 gradient. Add 2μL of the diluted compound to 78μL of 1x kinase buffer to obtain 2.5x target compound. Add 2μL of dimethyl sulfoxide (DMSO) to the positive control group and blank control group. 6) Preparation of 1 μM Streptavidin-XL665: The concentration of Streptavidin-XL665 is 16.67 μM, which can be diluted 16.67 times using the detection buffer. 7) Preparation of 1x TK-Antibody: The TK-Antibody stock solution is a 100x solution, which is diluted to 1x with detection buffer before use.

[0095] 3.2 Test Method 1) 4 μL of the 2.5× target compound was added to a 96-well HTRF microplate, followed by 2 μL of 5× substrate to one side of the well, 2 μL of 5× JAK1 to the other side of the well, and 2 μL of 1× kinase buffer to the blank control group. 2) The plate was sealed with a sealing plate film, placed in a centrifuge, and centrifuged at 1000 rpm for 2 minutes. 3) 2 μL of 5×ATP was added per well, the plate was sealed with a sealing plate film, centrifuged at 1000 rpm for 1 minute, and the plate was placed in an incubator at 30° C. and incubated for 4 hours. 4) After the incubation was completed, the above 1 μM Streptavidin-XL665 and 1×TK-Antibody were mixed at a 1:1 ratio, and 10 μL was added per well, followed by centrifugation at 1000 rpm for 1 minute. 5) The plate was returned to the incubator and incubated for another hour. After the incubation was completed, the HTRF665 / 620 signal was read on the multifunctional microplate reader.

[0096] 4.JAK2 kinase inhibitory activity test 4.1 Reagent preparation 1) Preparation of 1x kinase buffer: 5x kinase buffer was diluted with sterile water to 1x kinase buffer, and then 5mM MgCl2 and 1mM DTT were added. 2) Preparation of 5x JAK2: The concentration of the JAK2 mother solution was 166 ng / μL, and it was adjusted to 5x the final concentration, i.e., 0.04 ng / μL. The mother solution was first diluted to 1.66 ng / μL, and then diluted 41.5 times from 1.66 ng / μL to prepare 0.04 ng / μL. 3) Preparation of 5x ATP: 4 µM ATP was prepared at 5x its concentration, i.e., 20 µM, and directly diluted 500-fold from 10 mM ATP to obtain the required ATP concentration. 4) Preparation of 5x substrate: 5x of 1µM, i.e. 5µM, the substrate concentration was 500µM, which was diluted to 100x to give 5µM substrate. 5) Preparation of 2.5x target compound: The concentration of the compound buffer solution was 10mM. The treatment concentration started from 10μM. First, the compound was prepared in 100x buffer solution, i.e., 1mM, and then diluted from 10mM to 10x. Then, the diluted compound was diluted in a 1:3 gradient to a total of 10 concentrations. 2μL of the diluted compound was added to 78μL of 1x kinase buffer solution to obtain 2.5x target compound. 2μL of dimethyl sulfoxide (DMSO) was added to the positive control group and the blank control group. 6) Preparation of 1 μM Streptavidin-XL665: The concentration of Streptavidin-XL665 is 16.67 μM, which can be diluted 16.67 times using the detection buffer. 7) Preparation of 1x TK-Antibody: The TK-Antibody stock solution is a 100x solution, which can be diluted to 1x with detection buffer before use.

[0097] 4.2 Test Methodology 1) 4 μL of the above 2.5× target compound was added to an HTRF 96-well microplate, followed by 2 μL of 5× substrate to one side of the well, 2 μL of 5× JAK2 to the other side of the well, and 2 μL of 1× kinase buffer to the blank control group. 2) The plate was sealed with a sealing plate film, placed in a centrifuge, and centrifuged at 1000 rpm for 2 minutes. 3) 2 μL of 5×ATP was added per well, the plate was sealed with a sealing plate film, centrifuged at 1000 rpm for 1 minute, and the plate was placed in an incubator at 30° C. and incubated for 2 hours. 4) After the incubation was completed, the above 1 μM Streptavidin-XL665 and 1×TK-Antibody were mixed at a 1:1 ratio, and 10 μL was added per well, followed by centrifugation at 1000 rpm for 1 minute. 5) The plate was returned to the incubator and incubated for another hour. After the incubation was completed, the HTRF665 / 620 signal was read on the multifunctional microplate reader.

[0098] 5.JAK3 kinase inhibitory activity test 5.1 Reagent preparation 1) Preparation of 1x kinase buffer: 5x kinase buffer was diluted with sterile water to 1x kinase buffer, and then 5mM MgCl2 and 1mM DTT were added. 2) Preparation of 5xJAK3: The concentration of the JAK3 mother solution was 124 ng / μL, and it was adjusted to 5x the final concentration, i.e., 0.5 ng / μL, and the mother solution was diluted 248-fold to obtain the required JAK3 concentration. 3) Preparation of 5x ATP: 3 µM ATP was prepared at 5x its concentration, i.e., 15 µM, and diluted 666.67 times from 10 mM ATP to obtain the required ATP concentration. 4) Preparation of 5x substrate: 5x of 1µM, i.e. 5µM, the substrate concentration was 500µM, which was diluted to 100x to make 5µM substrate. 5) Preparation of 2.5x target compound: The concentration of the compound buffer was 10mM, and the treatment concentration started from 10μM. First, prepare 100x buffer, i.e., 1mM, and then dilute from 10mM to 10x. Then, dilute to a total of 10 concentrations with a 1:3 gradient. Add 2μL of the diluted compound to 78μL of 1x kinase buffer to obtain 2.5x target compound. Add 2μL of dimethyl sulfoxide (DMSO) to the positive control group and blank control group. 6) Preparation of 1 μM Streptavidin-XL665: The concentration of Streptavidin-XL665 is 16.67 μM, and it is diluted 16.67 times with the detection buffer solution before use. 7) Preparation of 1x TK-Antibody: The TK-Antibody stock solution is a 100x solution, which can be diluted to 1x with detection buffer before use.

[0099] 5.2 Testing Methodology 1) 4 μL of the 2.5× target compound was added to a 96-well HTRF microplate, followed by 2 μL of 5× substrate to one side of the well, 2 μL of 5× JAK3 to the other side of the well, and 2 μL of 1× kinase buffer to the blank control group. 2) The plate was sealed with a sealing plate film, placed in a centrifuge, and centrifuged at 1000 rpm for 2 minutes. 3) 2 μL of 5×ATP was added per well, the plate was sealed with a sealing plate film, centrifuged at 1000 rpm for 1 minute, and the plate was placed in an incubator at 30° C. and incubated for 3 hours. 4) After the incubation was completed, the above 1 μM Streptavidin-XL665 and 1×TK-Antibody were mixed at a 1:1 ratio, and 10 μL was added per well, followed by centrifugation at 1000 rpm for 1 minute. 5) The plate was returned to the incubator and incubated for another hour. After the incubation was completed, the HTRF665 / 620 signal was read on the multifunctional microplate reader.

[0100] 6. Calculation of Inhibition Rate and IC 50 Fitting Inhibition rate = (positive control group - compound to be detected) / (positive control group - blank control group) x 100% The median inhibitory concentration (IC) was calculated using GraphPad Prism 6 according to the inhibitory rate of the target compound against the kinase at different concentrations. 50 ) was fitted.

[0101] The inhibitory activity of the representative compounds of the present application and the known control drug Filgotinib against JAK1, JAK2, and JAK3 kinases was measured by the above tests, and the measured IC 50 The values ​​are as shown in Table 1.

[0102] [Table 1]

[0103] As can be seen from Table 1, compared to the known control drug (Filgotinib), the compounds of the present application have good JAK1, JAK2 or JAK3 kinase inhibitory activity, and the compounds of the present application have clear selectivity against JAK1, JAK2 or JAK3 kinase.

[0104] Example 28: Pharmacokinetic testing 1. Experimental Animals Three healthy male C57 mice, 6–8 weeks old, were purchased from Shanghai Sino-British Sippr / BKLab Animal Co., Ltd.

[0105] 2. Experimental Method Mice were fasted overnight before oral dosing. Food was resumed 4 hours after dosing and water was allowed ad libitum. Mice were intragastrically administered 10 mg compound / kg body weight. Whole blood samples were collected via semi-continuous facial vein sampling. Approximately 30 μL of blood was collected at 0.125, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after dosing. The samples were placed in tubes containing the anticoagulant sodium heparin and placed on ice until centrifugation within 15 minutes. The samples were then centrifuged at 6800 g for 6 minutes at 6-8°C. Plasma was transferred to labeled tubes within 1 hour of collection and centrifugation and stored frozen at approximately -80°C.

[0106] 3. Chromatography and Mass Spectrometry Conditions The chromatography column is Luna (登録商標) The Omega ACQUITY UPLC BEH C18 (2.1 × 50 mm, 1.7 μm) was used. Mobile phase A was HO-0.1% FA, and mobile phase B was ACN-0.1% FA. The flow rate was 0.80 mL / min. The gradient elution procedure was 10% B at the start, 10% B at 0.6 min, 90% B at 1.0 min, 90% B at 1.11 min, and 10% B at 1.40 min. The column temperature was 40°C, and the injection volume was 2 μL. Mass spectrometry was performed using an LC-MS / MS-19 (TQ5500) (SCIEX, USA), with an ESI ion source, positive ion detection, and multiple reaction monitoring (MRM) scanning. m / z: 271.10 / 172.00 Da (tolbutamide, internal standard).

[0107] 4. Plasma Sample Preparation Ten microliters of plasma sample was taken, 200 μL of internal standard working solution (tolbutamide, 100 ng / mL) was added, vortexed for 1 minute, and centrifuged at 18,000 g for 10 minutes. 200 μL of the supernatant was transferred to a 96-well microplate, and 1 μL of the supernatant was injected into LC-MS / MS analysis.

[0108] 5. Analyzing the Results Pharmacokinetic (PK) parameters were calculated using Phoenix WinNonlin 7.0 software, and AUC, C max , T max The pharmacokinetic parameters of oral administration to mice, including T1 / 2, etc., were estimated, and the results of oral administration to mice of the representative compound 15 of the present application and the known control drug filgotinib are shown in Table 2.

[0109] [Table 2]

[0110] As can be seen from Table 2, compared to the known control drug (Filgotinib), the compound 15 of the present application at a unit dose has a higher systemic exposure (AUC), thus providing a clear pharmacokinetic advantage.

[0111] Although the preferred embodiments of the present application have been described in detail above, the present application is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solutions of the present application within the technical concept of the present application, and all of these simple modifications fall within the scope of protection of the present application.

[0112] It should be noted that the specific technical features described in the above specific embodiments can be combined in any suitable manner as long as there is no contradiction, and in this application, various possible combination manners will not be separately described in order to avoid unnecessary duplication.

[0113] Furthermore, various different embodiments of the present application can be arbitrarily combined, and should be considered as the contents disclosed in the present application in the same manner, unless it is contrary to the spirit of the present application. [Industrial Applicability]

[0114] The present application provides cyclopropanamide-containing compounds and methods for preparing and using the same. The cyclopropanamide-containing compounds provided herein have good inhibitory activity and clear selectivity against JAK1, JAK2, or JAK3 kinase, as well as favorable pharmacokinetic characteristics, and are of great medical value and market potential for diseases associated with abnormalities in the JAK signaling pathway (e.g., autoimmune diseases and myeloproliferative neoplasms).

Claims

1. A compound represented by general formula (III), its stereoisomer, or a pharmaceutically acceptable salt thereof. 【Transformation 3】 [In the formula, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; 【change】 R １ and R ２ are each independently hydrogen, halogen, or C １ ~C ６ Alkyl group, C １ ~C ６ haloalkyl group or C ３ ~C ７ cycloalkyl groups, R' is a substituted or unsubstituted C １ ~C ６ Alkyl group or C ３ ~C ７ cycloalkyl groups, the substitution groups being selected from halogen, C １ ~C ６ Alkoxy group, C １ ~C ６ Alkylthio group, C ３ ~C ７ Cycloalkyl group, hydroxy group, amino group, NHC １ ~C ６ Alkyl groups and N(C １ ~C ６ alkyl group) ２ is selected from the group consisting of:

2. 2. A compound represented by general formula (III) according to claim 1, having a structure represented by general formula (VII), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. 【Transformation 7】 [In the formula, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; R １ and R ２ are each independently hydrogen, halogen, or C １ ~C ６ Alkyl group, C １ ~C ６ haloalkyl group or C ３ ~C ７ cycloalkyl groups.

3. 2. The compound represented by general formula (III) according to claim 1, having a structure represented by general formula (VIII), its stereoisomer or a pharmaceutically acceptable salt thereof. 【Transformation 8】 [In the formula, X is selected from —CH or N; Y is selected from hydrogen or halogen, and n is 1 or 2; R' is a substituted or unsubstituted C １ ~C ６ Alkyl group or C ３ ~C ７ cycloalkyl groups, the substitution groups being selected from halogen, C １ ~C ６ Alkoxy group, C １ ~C ６ Alkylthio group, C ３ ~C ７ Cycloalkyl group, hydroxy group, amino group, NHC １ ~C ６ Alkyl groups and N(C １ ~C ６ alkyl group) ２ is selected from the group consisting of:

4. The compound represented by general formula (III) according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein Y is selected from H or F.

5. In the general formula The compound represented by general formula (III) according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, selected from the following:

6. In the general formula (VII), R １ and R ２ are each independently selected from hydrogen, fluorine, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a t-butyl group, an s-butyl group, an n-pentyl group, a trifluoromethyl group, a cyclopropyl group, and a cyclopentyl group, a compound represented by general formula (III), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to claim 2.

7. In the general formula (VIII), R' is selected from the group consisting of substituted or unsubstituted methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, s-butyl, n-pentyl, isopentyl, cyclopropyl, and cyclopentyl groups, and the substituents are selected from the group consisting of halogen, C １ ~C ６ Alkoxy group, C １ ~C ６ Alkylthio group, C ３ ~C ７ Cycloalkyl group, hydroxy group, amino group, NHC １ ~C ６ Alkyl groups and N(C １ ~C ６ alkyl group) ２ 4. The compound represented by general formula (III) according to claim 3, its stereoisomer or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

8. A compound represented by the following formula, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

9. A compound represented by the following formula, its stereoisomer, or a pharmaceutically acceptable salt thereof:

10. A compound represented by the following formula, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

11. A compound represented by the following formula, its stereoisomer, or a pharmaceutically acceptable salt thereof:

12. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 11, its stereoisomer or a pharmaceutically acceptable salt component thereof, and a pharmaceutically acceptable carrier.

13. Use of the compound according to any one of claims 1 to 11, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 12 in the manufacture of a medicament for treating a disease associated with an abnormality in the JAK signaling pathway.

14. The use according to claim 13, wherein the disease is an autoimmune disease.

15. The use of claim 14, wherein the autoimmune disease is selected from rheumatoid arthritis, ulcerative colitis, atopic dermatitis, or systemic lupus erythematosus.

16. The use according to claim 13, wherein the disease is a myeloproliferative neoplasm.

17. The use described in claim 16, wherein the myeloproliferative tumor-like disease is selected from essential thrombocytosis, polycythemia vera, or primary myelofibrosis.

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