A compound containing a gem-difluoro group, and a method of preparing and use thereof

Abstract

The present application provides a kind of compound shown in formula (I), its chiral enantiomer or its pharmaceutically acceptable salt and preparation method thereof, intermediate for preparing the compound shown in formula (I), its chiral enantiomer or its pharmaceutically acceptable salt and preparation method thereof, pharmaceutical composition comprising the compound shown in formula (I), its chiral enantiomer or its pharmaceutically acceptable salt and medical use of the compound shown in formula (I), its chiral enantiomer or its pharmaceutically acceptable salt or its pharmaceutical composition.The compound of the present application has good JAK2 kinase inhibitory activity, and it has higher selectivity to JAK2 kinase;In addition, the compound of the present application has obvious pharmacokinetic advantage, provides more choices for the prevention and / or treatment of related diseases of JAK signal pathway abnormality, and has better application prospect in clinic.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a compound containing a geminidinium fluoride group, its preparation method, and its uses. Background Technology

[0002] Janus kinases (JAKs) are a class of tyrosine kinases. The JAK family includes four members: JAK1, JAK2, JAK3, and TYK2. JAKs play important roles in the signal transduction of various cytokines.

[0003] Signal transducer and activator of transcription (STAT) is a group of cytoplasmic proteins that bind to the DNA of the regulatory regions of target genes. As downstream substrates of JAKs, STATs can be activated by tyrosine phosphorylation in response to external signals, and then translocate into the nucleus to regulate gene transcription.

[0004] Many abnormal immune responses, such as allergies, asthma, allogeneic transplant rejection, rheumatoid arthritis, and multiple sclerosis, as well as hematologic malignancies such as myelodysplastic syndromes, leukemia, and lymphoma, are related to abnormal regulation of the JAK / STAT signaling pathway.

[0005] JAK protein kinase inhibitors, especially JAK3 protein kinase inhibitors, can prevent T cell activation, prevent transplant rejection after transplantation, and also show efficacy against autoimmune diseases such as rheumatoid arthritis and multiple sclerosis. Furthermore, studies have found that myeloproliferative neoplasms (including essential thrombocythemia, polycythemia vera, and primary myelofibrosis) are associated with abnormal JAK2 kinase activity due to mutations. Therefore, the development of JAK2 protein kinase inhibitors has significant medical value and market potential for the treatment of myeloproliferative neoplasms.

[0006] Ruxolitinib is a selective JAK1 / 2 kinase inhibitor that was approved by the FDA in the United States on November 16, 2011, and marketed under the brand name Jakafi. It was the first drug in the United States to treat primary myelofibrosis (PMF). However, it lacks high selectivity against JAK2 kinase, and has a short half-life and low exposure.

[0007] Therefore, there is still a need to develop new compounds with high selectivity for JAK2 kinase and better pharmacokinetics. Summary of the Invention

[0008] The present invention aims to provide a compound of formula (I), its chiral enantiomer or a pharmaceutically acceptable salt thereof, and a method for preparing the same; an intermediate for preparing the compound of formula (I), its chiral enantiomer or a pharmaceutically acceptable salt thereof, and a method for preparing the same; a pharmaceutical composition comprising the compound of formula (I), its chiral enantiomer or a pharmaceutically acceptable salt thereof; and the pharmaceutical use of the compound of formula (I), its chiral enantiomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. The compounds of the present invention exhibit good JAK2 kinase inhibitory activity and high selectivity for JAK2 kinase. Furthermore, the compounds of the present invention have significant pharmacokinetic advantages, providing more options for the prevention and / or treatment of diseases related to abnormal JAK signaling pathways (especially autoimmune diseases, myeloproliferative neoplasms, and graft-versus-host disease), and have good clinical application prospects.

[0009] In a first aspect, the present invention provides compounds of formula (I), their chiral enantiomers, or pharmaceutically acceptable salts thereof:

[0010]

[0011] In formula (I),

[0012] A is an alkyl or cycloalkyl group, wherein the alkyl or cycloalkyl group is optionally substituted with fluorine, an alkyl group or a cycloalkylene group, and the alkyl, the cycloalkyl or the cycloalkylene group has at least one pair of geminofluorine groups;

[0013] X is H or (CH2) n Where n = 0, 1, 2, 3, 4 or 5; when X is (CH2) n When it is connected to A, it forms a C3-C7 cycloalkylene group, which is optionally substituted with fluorine or an alkyl group, and the cycloalkylene group or the alkyl group has at least one pair of geminofluorine groups.

[0014] In a preferred embodiment, the present invention provides the compound of formula (II), its chiral enantiomer, or a pharmaceutically acceptable salt thereof:

[0015]

[0016] In formula (II),

[0017] Y1 is CR1, where R1 is a bond, H, or F;

[0018] Z represents a bond or (CH2). m Where m = 1, 2 or 3;

[0019] Y2 is an alkylene or cycloalkylene, wherein the alkylene or cycloalkylene is optionally substituted with fluorine, alkyl or cycloalkylene, or Y2 is connected to Z or Y1 to form a C3-C7 cycloalkylene;

[0020] X is H or (CH2) n Where n = 0, 1, 2, 3, 4 or 5; when X is (CH2) n When it is connected to Y1, Z or Y2, it forms a C3-C7 cycloalkylene group.

[0021] As a preferred specific implementation, in the above formula (II):

[0022] Y1 is CR1, where R1 is a bond;

[0023] Z represents a bond or (CH2). m Where m = 1, 2 or 3;

[0024] Y2 is an alkylene group;

[0025] X is (CH2) n , where n = 0, 1, 2, 3, 4 or 5;

[0026] Furthermore, X and Y1 are connected to form a C3-C7 cycloalkylene group, preferably a C6 cycloalkylene group.

[0027] As another preferred embodiment, in equation (II) above:

[0028] Y1 is CR1, where R1 is F;

[0029] Z represents a bond or (CH2). m Where m = 1, 2 or 3;

[0030] Y2 is an alkylene group, preferably a C1-C5 alkylene group, and more preferably a C2-C3 alkylene group;

[0031] X is H.

[0032] As another preferred embodiment, in equation (II) above:

[0033] Y1 is CR1, where R1 is F;

[0034] Z represents a bond or (CH2). m Where m = 1, 2 or 3;

[0035] Y2 is an alkylene group;

[0036] X is (CH2) n , where n = 1, 2, 3, 4 or 5;

[0037] Furthermore, X and Z are connected to form a C3-C7 cycloalkylene group, preferably a C6 cycloalkylene group.

[0038] As another preferred embodiment, in equation (II) above:

[0039] Y1 is CR1, where R1 is a bond;

[0040] Z represents a bond or (CH2). m Where m = 1, 2 or 3;

[0041] Y2 is an alkylene group, and Y2 is connected to Y1 to form a C3-C7 cycloalkylene group, preferably a C4 or C6 cycloalkylene group;

[0042] X is H.

[0043] As another preferred embodiment, in equation (II) above:

[0044] Y1 is CR1, where R1 is F;

[0045] Z is (CH2) m Where m = 1, 2 or 3;

[0046] Y2 is an alkylene group, and Y2 is attached to Z to form a C3-C7 cycloalkylene group, preferably a C6 cycloalkylene group;

[0047] X is H.

[0048] As another preferred embodiment, in equation (II) above:

[0049] Y1 is CR1, where R1 is F;

[0050] Z represents a bond or (CH2). m Where m = 1, 2 or 3;

[0051] Y2 is an alkylene or cycloalkylene, which may optionally be substituted with an alkyl, cycloalkyl or cycloalkylene;

[0052] X is H.

[0053] In a preferred embodiment, the present invention provides compounds of the following formula, their chiral enantiomers, or pharmaceutically acceptable salts thereof:

[0054]

[0055]

[0056] In a second aspect, the present invention provides a method for preparing the compound according to the first aspect above, its chiral enantiomer, or a pharmaceutically acceptable salt thereof, comprising the following steps:

[0057] (1) Compound III-1 reacts with diethyl cyanomethyl phosphate in the presence of a base to generate compound III-2 via an olefination reaction;

[0058] (2) Compound III-2 and compound III-3 undergo a Mac addition reaction in the presence of a base and under heating conditions to generate compound III;

[0059] (3) Compound III undergoes deprotection in the presence of a base to form compound I;

[0060]

[0061] In a preferred embodiment, the alkali in step (1) is selected from any one or more of sodium hydride, sodium tert-butoxy, potassium tert-butoxy, lithium tert-butoxy, lithium bromide, lithium chloride, triethylamine, 4-dimethylaminopyridine, cesium carbonate, potassium carbonate, and sodium carbonate; the solvent in step (1) is an aprotic solvent, preferably selected from any one or more of tetrahydrofuran, dimethylformamide, and dimethyl sulfoxide;

[0062] The base mentioned in step (2) is selected from any one or more of 1,8-diazabicycloundec-7-ene, sodium tert-butoxy, potassium tert-butoxy, triethylamine, 4-dimethylaminopyridine, cesium carbonate, potassium carbonate, sodium carbonate, potassium phosphate, and sodium phosphate; the solvent mentioned in step (2) is a protic or aprotic solvent, preferably selected from any one or more of acetonitrile, N-methylpyrrolidone, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, methanol, and ethanol; the heating temperature mentioned in step (2) is 52-82℃;

[0063] The alkali mentioned in step (3) is selected from one or more of sodium hydroxide, lithium hydroxide and potassium hydroxide.

[0064] Thirdly, the present invention provides an intermediate for preparing the compound according to the first aspect above, its chiral enantiomers or pharmaceutically acceptable salts thereof, having a structure as shown in general formula (III):

[0065]

[0066] In formula (III),

[0067] A is an alkyl or cycloalkyl group, wherein the alkyl or cycloalkyl group is optionally substituted with fluorine, an alkyl group or a cycloalkylene group, and the alkyl, the cycloalkyl or the cycloalkylene group has at least one pair of geminofluorine groups;

[0068] X is H or (CH2) n Where n = 0, 1, 2, 3, 4 or 5; when X is (CH2)n When it is connected to A, it forms a C3-C7 cycloalkylene group, which is optionally substituted with fluorine or an alkyl group, and the cycloalkylene group or the alkyl group has at least one pair of geminofluorine groups.

[0069] Fourthly, the present invention provides a method for preparing the intermediate according to the third aspect above, comprising the following steps:

[0070] (1) Compound III-1 reacts with diethyl cyanomethyl phosphate in the presence of a base to generate compound III-2 via an olefination reaction;

[0071] (2) Compound III-2 and compound III-3 undergo a Mac addition reaction in the presence of a base and under heating conditions to generate compound III;

[0072]

[0073] In a preferred embodiment, the alkali in step (1) is selected from any one or more of sodium hydride, sodium tert-butoxy, potassium tert-butoxy, lithium tert-butoxy, lithium bromide, lithium chloride, triethylamine, 4-dimethylaminopyridine, cesium carbonate, potassium carbonate, and sodium carbonate; the solvent in step (1) is an aprotic solvent, preferably selected from any one or more of tetrahydrofuran, dimethylformamide, and dimethyl sulfoxide;

[0074] The base mentioned in step (2) is selected from any one or more of 1,8-diazabicycloundec-7-ene, sodium tert-butoxy, potassium tert-butoxy, triethylamine, 4-dimethylaminopyridine, cesium carbonate, potassium carbonate, sodium carbonate, potassium phosphate, and sodium phosphate; the solvent mentioned in step (2) is a protic or aprotic solvent, preferably selected from any one or more of acetonitrile, N-methylpyrrolidone, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, methanol, and ethanol; the heating temperature mentioned in step (2) is 52-82℃.

[0075] Fifthly, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the compound according to the first aspect above, its chiral enantiomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent and / or excipient.

[0076] In a sixth aspect, the present invention provides the use of the compounds according to the first aspect above, their chiral enantiomers or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions according to the fifth aspect above, in the preparation of medicaments for the prevention and / or treatment of diseases related to abnormalities of the JAK signaling pathway.

[0077] Preferably, the disease is an autoimmune disease, a myeloproliferative neoplasm, or a graft-versus-host disease.

[0078] More preferably, the autoimmune disease is selected from rheumatoid arthritis, ulcerative colitis, systemic lupus erythematosus, atopic dermatitis, or multiple sclerosis.

[0079] More preferably, the myeloproliferative neoplasm is selected from essential thrombocythemia, myelofibrosis, or polycythemia vera.

[0080] More preferably, the graft-versus-host disease is selected from acute graft-versus-host disease or chronic graft-versus-host disease.

[0081] Beneficial effects

[0082] 1. The compounds of the present invention have good JAK2 kinase inhibitory activity and high selectivity for JAK2 kinase;

[0083] 2. The compounds of this invention have significant pharmacokinetic advantages, providing more options for the prevention and / or treatment of diseases related to JAK signaling pathway abnormalities, and have good application prospects in clinical practice. Detailed Implementation

[0084] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions in the embodiments of this invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this invention, not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0085] Furthermore, to better illustrate the present invention, numerous specific details are provided in the following detailed embodiments. Those skilled in the art should understand that the present invention can be practiced without certain specific details. In some embodiments, materials, methods, etc., well-known to those skilled in the art, are not described in detail in order to highlight the spirit of the invention.

[0086] Unless otherwise expressly stated, throughout the specification and claims, the term "comprising" or its variations such as "including" or "comprises" shall be understood to include the stated components without excluding other components.

[0087] It should be noted that, unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0088] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups with 1 to 20 carbon atoms. Alkyl groups containing 1 to 10 carbon atoms are preferred, alkyl groups containing 1 to 6 carbon atoms are more preferred, alkyl groups containing 1 to 4 carbon atoms are most preferred, and methyl is the most desirable. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-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, 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 their various branched isomers, etc. More preferably, lower alkyl groups containing 1 to 6 carbon atoms, and non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-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, etc. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any usable connection point. The substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, amino, haloalkyl, hydroxyalkyl, carboxyl, or carboxylic acid ester.

[0089] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon group comprising 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, most preferably 3 to 6 carbon atoms, and most preferably cyclopropyl or cyclopentyl. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclohepttrienyl, cyclooctyl, etc., with cyclopropyl and cyclopentyl being preferred. Polycyclic cycloalkyl groups include spirocyclic, fused-ring, and bridged-ring cycloalkyl groups. The cycloalkyl group may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, amino, haloalkyl, hydroxyalkyl, carboxyl or carboxylic ester.

[0090] The term "alkylene" refers to a divalent, straight-chain or branched alkane group composed of carbon and hydrogen atoms, without unsaturation, and linked to one group via a single bond and to another group (or ring system) via a second single bond. For example, "C" as used in this text... 1-5 "alkylene" refers to an alkylene group containing 1-5 carbon atoms, "C 2-3 "Alkylene" refers to an alkylene containing 2-3 carbon atoms; non-limiting examples include methylene (-CH2-), 1,2-ethylene (-CH2CH2-), 1,3-propylene (-CH2CH2CH2-), 1-methyl-1,2-ethylene (-CH(CH3)CH2-), 1,4-butylene (-CH2CH2CH2CH2-), 1-methyl-1,3-propylene (-CH(CH3)CH2CH2-), 1,1-dimethyl-1,2-ethylene (-C(CH3)2CH2-), 1,2-dimethyl-1,2-ethylene (-CH(CH3)CH(CH3)-), etc.

[0091] The term "cycloalkylene" refers to a divalent monocyclic or polycyclic (including bridged and spirocyclic) non-aromatic cyclic hydrocarbon group consisting only of carbon and hydrogen atoms, without unsaturation, and linked to one group by a single bond and to other groups by another single bond. For example, "C" as used in this text... 3-7 "Cycloalkylene" refers to a cycloalkylene containing 3-7 carbon atoms, "C4 cycloalkylene" refers to a cycloalkylene containing 4 carbon atoms, and "C6 cycloalkylene" refers to a cycloalkylene containing 6 carbon atoms; non-limiting examples include cyclopropane-1,1-ylene, cyclopropane-1,2-ylene, cyclobutane-1,1-ylene, cyclobutane-1,2-ylene, cyclobutane-1,3-ylene, etc.

[0092] The term "bond" refers to the chemical bond between two atoms or two parts (i.e., groups or segments), where the atoms bonded by the bond are considered part of a larger substructure.

[0093] The term "chiral enantiomer" refers to two chiral molecules that are mirror images of each other and do not overlap.

[0094] The term "protecting group" refers to a group used to block the reactivity of a functional group. Examples of protecting groups include, but are not limited to, the methylene pivalate group.

[0095] The terms “optional” or “optionally” mean that the event or environment described below may, but does not have to, occur, and the description includes the possibility that the event or environment may or may not occur. For example, “optionally alkyl-substituted heterocyclic alkyl group” means that an alkyl group may, but does not have to, be present, and the description includes cases where the heterocyclic alkyl group is substituted with an alkyl group and cases where the heterocyclic alkyl group is not substituted with an alkyl group.

[0096] "Substituted" refers to one or more hydrogen atoms in a group, preferably up to five, more preferably one to three hydrogen atoms, which are independently substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom having an unsaturated bond (such as an alkene).

[0097] Unless otherwise stated, the instruments, consumables, and reagents used in the following examples are all available through conventional commercial means. Experimental methods for which specific conditions are not specified in the examples shall be performed in accordance with conventional methods and conditions or as per the product instructions.

[0098] Preparation of the compound and its intermediates as shown in formula (I) of this invention

[0099] The intermediate of the compound shown in formula (I) of this invention can be synthesized according to the following general synthetic route:

[0100]

[0101] Specifically, the following steps are included:

[0102] (1) Compound III-1 reacts with diethyl cyanomethyl phosphate in the presence of a base to generate compound III-2 via an olefination reaction;

[0103] Preferably, the alkali is selected from one or more of sodium hydride, sodium tert-butoxy, potassium tert-butoxy, lithium tert-butoxy, lithium bromide, lithium chloride, triethylamine, 4-dimethylaminopyridine, cesium carbonate, potassium carbonate, and sodium carbonate; preferably, the solvent used is selected from one or more of tetrahydrofuran, dimethylformamide, and dimethyl sulfoxide.

[0104] (2) Compound III-2 and compound III-3 undergo a Mac addition reaction in the presence of a base and under heating conditions to generate compound III;

[0105] Preferably, the alkali is selected from any one or more of 1,8-diazabicycloundec-7-ene, sodium tert-butoxy, potassium tert-butoxy, triethylamine, 4-dimethylaminopyridine, cesium carbonate, potassium carbonate, sodium carbonate, potassium phosphate, and sodium phosphate; preferably, the solvent is selected from any one or more of acetonitrile, N-methylpyrrolidone, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, methanol, and ethanol; the heating temperature is 52-82°C.

[0106] The compound represented by formula (I) of this invention can be synthesized according to the following general synthetic route:

[0107]

[0108] Specifically, the following steps are included:

[0109] (1) The preparation method of compound III-2 is as described above;

[0110] (2) The preparation method of compound III is as described above;

[0111] (3) Compound III is deprotected in the presence of a base to generate compound I; preferably, the base is selected from one or more of sodium hydroxide, lithium hydroxide and potassium hydroxide.

[0112] The following provides some specific preparation examples of the compound of formula (I) of the present invention, as well as its JAK2 / JAK3 kinase inhibitory activity test and pharmacokinetic test.

[0113] Example 1: Synthesis of Compound 1

[0114]

[0115] The specific steps are as follows:

[0116] Step (1):

[0117] Diethyl cyanomethyl phosphate (1.95 g, 11 mmol) was dissolved in dry tetrahydrofuran (50 mL). Sodium hydride (420 mg, 13 mmol) was added under ice bath conditions. After stirring for one hour, compound 1A (1.34 g, 10 mmol) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with saturated ammonium chloride solution (20 mL), extracted with ethyl acetate (20 mL × 3), and the organic phases were combined. The mixture was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue, which was compound 1B (LCMS (ESI+): 158.08 (M+H)). + ), which is used directly in the next step.

[0118] Step (2):

[0119] Compound 1C (1.35 g, 5 mmol), compound 1B (15 mmol) obtained in step (1), and 1,8-diazacyclic [5,4,0]undecene-7 (5 mmol) were dissolved in acetonitrile (30 mL) and reacted at 80 °C for five days. After returning to room temperature, most of the acetonitrile was removed by concentration, and water (20 mL) and ethyl acetate (10 mL × 3) were added for extraction. 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 purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to obtain compound 1D (LCMS(ESI+): 457.22 (M+H). + ).

[0120] Step (3):

[0121] Compound 1D (912 mg, 2 mmol) obtained in step (2) was dissolved in methanol and water (5 / 5 mL), and sodium hydroxide (160 mg, 4 mmol) was added. The reaction was carried out at room temperature and monitored by TLC until the starting material was consumed. 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 purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to obtain compound 1. (LCMS (ESI+): 343.15 (M+H)) + ).

[0122] Example 2: Synthesis and resolution of compound 2

[0123]

[0124] The specific steps are as follows:

[0125] Step (1):

[0126] Compound 2A (1.42 g, 10 mmol) was dissolved in dichloromethane (20 mL), silica gel powder (SiO2, 3 g) and pyridine chlorochromate (3.24 g, 15 mmol) were added, and the mixture was stirred overnight at room temperature. After filtration through diatomaceous earth, the solution was concentrated to obtain compound 2B (LCMS(ESI+): 141.05 (M+H)). + ), which is used directly in the next step.

[0127] Step (2):

[0128] Diethyl cyanomethyl phosphate (1.95 g, 11 mmol) was dissolved in dry tetrahydrofuran (50 mL). Sodium hydride (420 mg, 13 mmol) was added under ice bath conditions. After stirring for one hour, compound 2B (1.4 g, 10 mmol) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with saturated ammonium chloride solution (20 mL), extracted with ethyl acetate (20 mL × 3), and the organic phases were combined. The mixture was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue, which was compound 2C (LCMS (ESI+): 164.07 (M+H)). + ), which is used directly in the next step.

[0129] Step (3):

[0130] Compound 1C (1.35 g, 5 mmol), compound 2C (15 mmol) obtained in step (2), and 1,8-diazacyclic [5,4,0]undecene-7 (5 mmol) were dissolved in acetonitrile (30 mL) and reacted at 80 °C for five days. After returning to room temperature, most of the acetonitrile was removed by concentration, and water (20 mL) and ethyl acetate (10 mL × 3) were added for extraction. 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 purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to obtain compound 2D (LCMS(ESI+): 463.21 (M+H)). + ).

[0131] Step (4):

[0132] Compound 2D (924 mg, 2 mmol) obtained in step (3) was dissolved in methanol and water (5 / 5 mL), and sodium hydroxide (160 mg, 4 mmol) was added. The reaction was carried out at room temperature and monitored by TLC until the starting material was consumed. 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 purified by column chromatography (dichloromethane:methanol = 100:0-100:5) to obtain compound 2. (LCMS (ESI+): 349.14 (M+H)) + ).

[0133] Resolution of compound 2

[0134] Resolution conditions: Compound 2 was dissolved in ethanol (6 mg / mL). A chiral UniChiral CND-H column (50 mm I.D x 250 mm L) was used, with a mobile phase of n-hexane / ethanol = 80 / 20 (V / V), a flow rate of 120 mL / min, UV at 254 nm, and a column temperature of 30 °C. After resolution, compounds 2-1 and 2-2 were obtained, respectively.

[0135] Retention time: Compound 2-1: 8.493 min (>98% ee), Compound 2-2: 9.994 min (>98% ee).

[0136] Example 3, Synthesis of Compound 3

[0137]

[0138] Following the synthetic route of Example 1, compound 3 was obtained using 4-trifluoromethylcyclohexane-1-one as the starting material. LCMS (ESI+): 375.15 (M+H) + .

[0139] Example 4, Synthesis of Compound 4

[0140]

[0141] Following the synthetic route of Example 1, compound 4 was obtained using 4,4-difluorocyclohexylcarboxaldehyde as the starting material. LCMS (ESI+): 357.16 (M+H) + .

[0142] Example 5: Synthesis of Compound 5

[0143]

[0144] Following the synthetic route of Example 2, compound 5 was obtained using 4-(trifluoromethyl)cyclohexylethanol as the starting material. LCMS (ESI+): 389.17 (M+H) + .

[0145] Example 6: Synthesis and resolution of compound 6

[0146]

[0147] Following the synthetic route of Example 1, compound 6 was obtained using 4,4,4-trifluorobutyraldehyde as the starting material. LCMS (ESI+): 335.12 (M+H) + .

[0148] Resolution of compound 6

[0149]

[0150] Resolution conditions: Compound 6 was dissolved in n-hexane / ethanol = 80 / 20 (V / V) (concentration 10 mg / mL). Chiral column ChiralCel OD-H (0.46 cm ID x 25 cm L), mobile phase n-hexane / ethanol = 80 / 20 (V / V), flow rate 1.0 mL / min, UV 254 nm, column temperature 30 °C.

[0151] Retention time: Compound 6-1: 9.295 min (>98% ee), Compound 6-2: 11.252 min (>98% ee).

[0152] Example 7, Synthesis of Compound 7

[0153]

[0154] Following the synthetic route of Example 2, compound 7 was obtained using (3,3-difluorocyclobutyl)methanol as the starting material. LCMS (ESI+): 329.13 (M+H) + .

[0155] Example 8, Synthesis of Compound 8

[0156]

[0157] Following the synthetic route of Example 2, compound 8 was obtained using 3,3,3-trifluoro-2,2-dimethylpropane-1-ol as the starting material. LCMS (ESI+): 349.14 (M+H) + .

[0158] Example 9, Synthesis of Compound 9

[0159]

[0160] Following the synthetic route of Example 2, compound 9 was obtained using 1-(trifluoromethyl)-1-cyclobutyl-1-methanol as the starting material. LCMS (ESI+): 361.14 (M+H) + .

[0161] Example 10: Synthesis of Compound 10

[0162]

[0163] Following the synthetic route of Example 2, compound 10 was obtained using 4,4,5,5,5-pentafluoropentanol as the starting material. LCMS (ESI+): 385.12 (M+H) + .

[0164] Example 11, Other Preparation Examples

[0165] Other specific examples of compound preparation include:

[0166] Table 1. Preparation and characterization of other compounds of the present invention.

[0167]

[0168] Example 12: JAK2 / JAK3 kinase inhibitory activity test

[0169] 1. Experimental consumables

[0170] JAK2: Carna 09-045 14CBS-0374H

[0171] JAK3: Carna 08-046 19CBS-0798B

[0172] ATP (10mM): CST 9804

[0173] DTT: 100mM

[0174] MgCl2: 1M

[0175] TK substrate-biotin (biotin-labeled tyrosine kinase substrate, hereinafter referred to as substrate): Cisbio, #61TK0BLC*

[0176] Streptavidin-XL665 (XL665 with streptavidin label): Cisbio, #610SAXLG*

[0177] HTRF Kinase-TK kit: Cisbio, #62TK0PEC

[0178] TK-Antibody-Eu 3 -Cryptate (Europium-labeled tyrosine kinase antibody): From kit Cisbio, #62TK0PEC

[0179] HTRF 96-well low volume plate: Cisbio, #66PL96001

[0180] Ruxolitinib: Chongqing Gongzhou Pharmaceutical Co., Ltd., CAS No.: 941678-49-5

[0181] 2. Test conditions

[0182] JAK2: 0.008 ng / μL, ATP 4 μM, substrate 1 μM, time 2 h

[0183] JAK3: 0.1 ng / μL, ATP 3 μM, substrate 1 μM, time 3 h

[0184] 3. JAK2 kinase inhibitory activity test

[0185] 3.1 Reagent Preparation

[0186] 1) Preparation of 1× kinase buffer: Dilute 5× kinase buffer with sterile water to 1× kinase buffer, then add 5mM MgCl2 and 1mM DTT;

[0187] 2) Preparation of 5× JAK2: The JAK2 concentration is 166 ng / μL. To prepare a final concentration of 5×, i.e. 0.04 ng / μL, first dilute to 1.66 ng / μL, then dilute 1.66 ng / μL 41.5 times to prepare 0.04 ng / μL.

[0188] 3) 5×ATP preparation: 4μM ATP is prepared to be 5×, i.e. 20μM. The required ATP concentration is obtained by directly diluting 10mM ATP 500 times.

[0189] 4) 5× substrate preparation: 1 μM of 5× is equivalent to 5 μM, and the substrate concentration is 500 μM. Diluting it by 100× will give you 5 μM of substrate.

[0190] 5) Preparation of 2.5× test compound: The concentration of the test compound buffer is 10 mM. The treatment concentration starts at 10 μM. First, prepare a 100× buffer, i.e., 1 mM. Dilute 10 mM to 10×, and then perform serial dilutions at a ratio of 1:3, for a total of 10 concentrations. Take 2 μL to 78 μL of the diluted test compound solution and add it to 1× kinase buffer, which is the 2.5× test compound.

[0191] In addition, add 2 μL of DMSO to 78 μL of 1× kinase buffer to obtain 2.5× DMSO;

[0192] 6) Preparation of 1μM Streptavidin-XL665: The concentration of Streptavidin-XL665 is 16.67μM. When using, dilute it 16.67 times with detection buffer.

[0193] 7) 1×TK-Antibody-Eu 3 -Cryptate formulation: TK-Antibody-Eu 3 -Cryptate stock solution is a 100× solution; dilute to 1× with detection buffer before use.

[0194] 3.2 Test Method

[0195] 1) T-compound preparation: Add 4 μL of the above 2.5× analyte to the HTRF 96-well microplate, then add 2 μL of 5× substrate to one side of the well and 2 μL of 5× JAK2 to the other side of the well;

[0196] DMSO control wells without analyte (T-enzyme): Add 4 μL of the above 2.5×DMSO to an HTRF 96-well microplate, then add 2 μL of 5×substrate to one side of the well and 2 μL of 5×JAK2 to the other side of the well;

[0197] Enzyme-free blank control (T-without enzyme): Add 4 μL of the above 2.5×DMSO to an HTRF 96-well microplate, then add 2 μL of 5× substrate to one side of the well and 2 μL of 1× kinase buffer to the other side of the well.

[0198] 2) Seal the plate with sealing film, put it in a centrifuge, and centrifuge at 1000 rpm for 2 minutes;

[0199] 3) Add 2 μL of 5×ATP to each well, seal with sealing film, centrifuge at 1000 rpm for 1 minute, and incubate the plate in a 30℃ incubator for 2 hours;

[0200] 4) After incubation, combine the above-mentioned Streptavidin-XL665 and 1×TK-Antibody-Eu 3 Mix the Cryptotate at a 1:1 ratio, add 10 μL to each well, and centrifuge at 1000 rpm for 1 minute;

[0201] 5) Place the plate back into the incubator and continue incubation for 1 hour. After incubation, read the HTRF 620 / 665 signal on a multi-functional microplate reader.

[0202] 4. JAK3 kinase inhibitory activity test

[0203] 4.1 Reagent Preparation

[0204] 1) Preparation of 1× kinase buffer: Dilute 5× kinase buffer with sterile water to 1× kinase buffer, then add 5mM MgCl2 and 1mM DTT;

[0205] 2) Preparation of 5× JAK3: The JAK3 concentration is 124 ng / μL. To prepare it to the final concentration of 5×, which is 0.5 ng / μL, dilute it 248 times to get the required concentration.

[0206] 3) 5×ATP preparation: 3μM ATP is prepared to be 5×, i.e. 15μM. The required ATP concentration is obtained by directly diluting 10mM ATP by 666.67 times.

[0207] 4) 5× substrate preparation: 1 μM of 5× is equivalent to 5 μM, and the substrate concentration is 500 μM. Diluting it by 100× will give you 5 μM of substrate.

[0208] 5) Preparation of 2.5× test compound: The concentration of the test compound buffer is 10 mM. The treatment concentration starts at 10 μM. First, prepare a 100× buffer, i.e., 1 mM. Dilute 10 mM to 10×, and then perform serial dilutions at a ratio of 1:3, for a total of 10 concentrations. Take 2 μL to 78 μL of the diluted test compound solution and add it to 1× kinase buffer, which is the 2.5× test compound.

[0209] In addition, add 2 μL of DMSO to 78 μL of 1× kinase buffer to obtain 2.5× DMSO;

[0210] 6) Preparation of 1μM Streptavidin-XL665: The concentration of Streptavidin-XL665 is 16.67μM. When using, dilute it 16.67 times with detection buffer.

[0211] 7) 1×TK-Antibody-Eu 3 -Cryptate formulation: TK-Antibody-Eu 3-Cryptate stock solution is a 100× solution; dilute to 1× with detection buffer before use.

[0212] 4.2 Test Methods

[0213] 1) T-compound preparation: Add 4 μL of the above 2.5× analyte to the HTRF 96-well microplate, then add 2 μL of 5× substrate to one side of the well and 2 μL of 5× JAK3 to the other side of the well;

[0214] DMSO control wells without analyte (T-enzyme): Add 4 μL of the above 2.5×DMSO to an HTRF 96-well microplate, then add 2 μL of 5×substrate to one side of the well and 2 μL of 5×JAK3 to the other side of the well;

[0215] Enzyme-free blank control (T-without enzyme): Add 4 μL of the above 2.5×DMSO to an HTRF 96-well microplate, then add 2 μL of 5× substrate to one side of the well and 2 μL of 1× kinase buffer to the other side of the well.

[0216] 2) Seal the plate with sealing film, put it in a centrifuge, and centrifuge at 1000 rpm for 2 minutes;

[0217] 3) Add 2 μL of 5×ATP to each well, seal the plate with the sealing film, centrifuge at 1000 rpm for 1 minute, and incubate the plate in a 30℃ incubator for 3 hours;

[0218] 4) After incubation, combine the above-mentioned Streptavidin-XL665 and 1×TK-Antibody-Eu 3 Mix the Cryptotate at a 1:1 ratio, add 10 μL to each well, and centrifuge at 1000 rpm for 1 minute;

[0219] 5) Place the plate back into the incubator and continue incubation for 1 hour. After incubation, read the HTRF 620 / 665 signal on a multi-functional microplate reader.

[0220] 5. Inhibition rate calculation and IC50 fitting

[0221] Inhibition rate=(T-enzyme-T-compound) / (T-enzyme-T-without enzyme)×100%

[0222] Based on the inhibition rate of the test compound on the kinase at different concentrations, the half-maximal inhibitory concentration (IC50) was fitted using GraphPad Prism 6.

[0223] The inhibitory activity of the representative compound of this invention and the positive control drug ruxotinib against JAK2 and JAK3 kinases was tested using the above experiments, and the measured IC50 values ​​are shown in Table 2 below.

[0224] Table 2. IC50 values ​​of representative compounds of this invention and ruxolitinib for inhibiting the activity of JAK2 and JAK3 kinases.

[0225]

[0226] As shown in Table 2, the compounds of the present invention have good JAK2 kinase inhibitory activity, and the selectivity of the compounds of the present invention for JAK2 is equal to or better than that of the positive control drug ruxolitinib.

[0227] Example 13, Pharmacokinetic Test

[0228] 1. Laboratory animals

[0229] Three healthy male C57 mice, aged 6-8 weeks, were purchased from Shanghai Sippr / BKLab Animal Co., Ltd.

[0230] 2. Experimental Methods

[0231] Mice were fasted overnight before oral administration and given food and water freely 4 hours after administration. Mice were administered the compound at 10 mg / kg body weight via gavage. Whole blood samples were collected using a semi-continuous facial vein sampling method. Approximately 30 μL of blood was collected at 0.125 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, and 24 h after administration. The samples were placed in test tubes containing the anticoagulant sodium heparin and kept on ice until centrifugation. The plasma was centrifuged at 6800 g for 6 min at 6-8 °C within 15 min of the centrifuge. Within 1 hour of blood collection / centrifugation, the plasma was transferred to appropriately labeled test tubes and frozen at approximately -80 °C.

[0232] 3. Chromatographic and mass spectrometric conditions

[0233] chromatographic column is Omega ACQUITY UPLC BEH C18 (2.1×50mm, 1.7μm); Mobile phase A: H2O-0.1%FA, Mobile phase B: ACN-0.1%FA, Flow rate: 0.80mL / min; Gradient elution program: Initial, 10%B; 0.6min, 10%B; 1.0min, 90%B; 1.11min, 90%B; 1.40min, 10%B; Column temperature: 40℃, Injection volume: 2μL.

[0234] Mass spectrometry method: LC-MS / MS-19 (TQ5500) (SCIEX, USA), ion source: ESI source, detection mode: positive ion detection, scanning mode: multiple reaction monitoring (MRM), m / z: 271.10 / 172.00 Da (tolbutamide, internal standard).

[0235] 4. Plasma Sample Preparation

[0236] Take 10 μL of plasma sample, add 200 μL of internal standard working solution (tolbutamide, 100 ng / mL), vortex for 1 min, centrifuge at 18000 g for 10 min, transfer 200 μL of supernatant to a 96-well microplate, and inject 1 μL of supernatant into LC-MS / MS for analysis.

[0237] 5. Results Analysis

[0238] Pharmacokinetic (PK) parameters were calculated using Phoenix WinNonlin 7.0; oral pharmacokinetic parameters in mice, including AUC and C, were estimated using a non-compartmental model. max T max Table 3 shows the oral pharmacokinetic parameters of the representative compounds of the present invention (prepared from the above examples, wherein compound 2 is a mixture of compound 2-1 and compound 2-2) and the positive control drug ruxotinib in mice.

[0239] Table 3. Oral pharmacokinetic parameters of representative compounds of this invention and ruxotinib

[0240]

[0241] *dn refers to dose normalization.

[0242] Table 3 shows the half-life (T1 / 2) and exposure per unit dose (AUC) of the compounds of this invention. (0-∞) The dn values ​​were significantly higher than those of the positive control drug ruxotinib, thus demonstrating a clear pharmacokinetic advantage.

[0243] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A compound, a chiral enantiomer thereof, or a pharmaceutically acceptable salt thereof, characterized by, has a structure as shown in formula (II): (II) In formula (II), Y1 is CR1, wherein R1 is a bond; Z is a bond or (CH2) m wherein m = 1, 2 or 3; Y2 is alkylene; X is (CH2) n wherein n = 1, 2, 3, 4 or 5; and X is connected with Y1 to form C6 cyclic alkylene.

2. A compound, a chiral enantiomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, has a structure as shown in formula (II): (II) In formula (II), Y1 is CR1, wherein R1 is a bond; Z is a bond or (CH2) m wherein m = 1, 2 or 3; Y2 is C1-C3 alkylene; X is H.

3. A compound, a chiral enantiomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, has a structure as shown in formula (II): (II) In formula (II), Y1 is CR1, wherein R1 is a bond; Z is a bond or (CH2) m wherein m = 1, 2 or 3; Y2 is alkylene; X is (CH2) n wherein n = 1, 2, 3, 4 or 5; and X is connected with Z to form C6 cyclic alkylene.

4. A compound, a chiral enantiomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, has a structure as shown in formula (II): (II) In formula (II), Y1 is CR1, wherein R1 is a bond; Z is a bond or (CH2) m wherein m = 1, 2 or 3; Y2 is alkylene, and Y2 is connected with Y1 to form C4 cyclic alkylene; X is H.

5. A compound, a chiral enantiomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, has a structure as shown in formula (II): (II) In formula (II), Y1 is CR1, wherein R1 is a bond; Z is (CH2) m wherein m = 1, 2 or 3; Y2 is alkylene, and Y2 is connected with Z to form C4-C6 cyclic alkylene; X is H.

6. A compound of the following formula, a chiral enantiomer thereof, or a pharmaceutically acceptable salt thereof: 。 7. A method for preparing the compound, a chiral enantiomer thereof, or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, comprising the following steps: (1) compound III-1 is subjected to an ene reaction with cyanomethyl diethyl phosphate in the presence of a base in a solvent to generate compound III-2; (2) compound III-2 is subjected to a McCCaddition reaction with compound III-3 in the presence of a base and under heating conditions in a solvent to generate compound III; (3) compound III is deprotected in the presence of a base to generate compound I; ； wherein, A is Y1, Z, Y2 are as defined in any one of claims 1 to 6; and wherein X is defined as in any one of claims 1-6.

8. The method for preparing according to claim 7, wherein, in step (1), the base is selected from any one or several of sodium hydride, sodium tert-butoxide, potassium tert-butoxide, lithium tert-butoxide, lithium bromide, lithium chloride, triethylamine, 4-dimethylaminopyridine, cesium carbonate, potassium carbonate, and sodium carbonate; and the solvent is an aprotic solvent; in step (2), the base is selected from any one or several of 1,8-diazabicycloundec-7-ene, sodium tert-butoxide, potassium tert-butoxide, triethylamine, 4-dimethylaminopyridine, cesium carbonate, potassium carbonate, sodium carbonate, potassium phosphate, and sodium phosphate; the solvent is a protic or aprotic solvent; and the heating temperature of the heating conditions is 52-82°C; in step (3), the base is selected from any one or several of sodium hydroxide, lithium hydroxide, and potassium hydroxide.

9. The production method according to claim 8, characterized by, in step (1), the solvent is selected from any one or several of tetrahydrofuran, dimethylformamide, and dimethyl sulfoxide; in step (2), the solvent is selected from any one or several of acetonitrile, N-methylpyrrolidone, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, methanol, and ethanol.

10. An intermediate for preparing the compound, a chiral enantiomer thereof, or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, having a structure as shown in general formula (III): (III) in the formula (III), A is ; wherein, Y1, Z, Y2are as defined in any one of claims 1-6; and wherein X is as defined in any one of claims 1-6.

11. The method for preparing the intermediate according to claim 10, comprising the following steps: (1) compound III-1 is reacted with cyanomethyl diethyl phosphate in the presence of a base in a solvent to form compound III-2 by olefination; (2) compound III-2 is subjected to a McCCaddition reaction with compound III-3 in the presence of a base and under heating conditions in a solvent to form compound III; 。 12. The method for preparing according to claim 11, characterized in that, the base in step (1) is selected from any one or several of sodium hydride, sodium tert-butoxide, potassium tert-butoxide, lithium tert-butoxide, lithium bromide, lithium chloride, triethylamine, 4-dimethylaminopyridine, cesium carbonate, potassium carbonate and sodium carbonate; the solvent in step (1) is an aprotic solvent; the base in step (2) is selected from any one or several of 1,8-diazabicycloundec-7-ene, sodium tert-butoxide, potassium tert-butoxide, triethylamine, 4-dimethylaminopyridine, cesium carbonate, potassium carbonate, sodium carbonate, potassium phosphate and sodium phosphate; the solvent in step (2) is a protic or aprotic solvent; the heating temperature of the heating conditions in step (2) is 52-82°C.

13. The method of claim 12, wherein, the solvent in step (1) is selected from any one or several of tetrahydrofuran, dimethylformamide and dimethyl sulfoxide; the solvent in step (2) is selected from any one or several of acetonitrile, N-methylpyrrolidone, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, methanol and ethanol.

14. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1-6, a chiral enantiomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

15. The pharmaceutical composition of claim 14, wherein, The carrier is an excipient.

16. Use of a compound according to any one of claims 1-6, a chiral enantiomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to any one of claims 14-15 in the preparation of a medicament for preventing and / or treating a disease related to abnormal JAK signaling pathway; wherein, The disease is an autoimmune disease, a myeloproliferative neoplasm disease or a graft-versus-host disease.

17. Use according to claim 16, characterized in that, The autoimmune disease is selected from rheumatoid arthritis, ulcerative colitis, systemic lupus erythematosus, atopic dermatitis or multiple sclerosis; The myeloproliferative neoplasm disease is selected from essential thrombocythemia, myelofibrosis or polycythemia vera; The graft-versus-host disease is selected from acute graft-versus-host disease or chronic graft-versus-host disease.

Images