Pyrimidopyrrole compounds

Abstract

The present application provides a pyrimidine five-membered ring compound represented by formula (Ib) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition and a preparation method thereof, and use thereof as a JAK3 and / or BTK inhibitor.

Description

[0001] This application claims the priority of two prior applications, patent application number 202010071941.X, filed with the State Intellectual Property Office of China on January 21, 2020, with the invention name “Pyrimido-pyrrole compounds” and patent application number 202011003941.2, filed with the State Intellectual Property Office of China on September 22, 2020, with the invention name “Pyrimido-pyrrole compounds”. The full texts of the two applications are incorporated into this application by reference. Technical Field

[0002] The present invention relates to a novel pyrimidopyrrole compound or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the same and use thereof in preventing or treating kinase-related diseases such as Janus kinase (JAK, especially JAK3) and / or Bruton's tyrosine kinase (BTK)-related diseases. Background Art

[0003] Autoimmune diseases are a type of disease caused by abnormal immune function that attacks one's own cells or tissues, leading to inflammation and tissue damage, including rheumatoid arthritis (RA), inflammatory bowel disease (IBD) and systemic lupus erythematosus (SLE). BTK and JAK3 are two important targets for autoimmune diseases.

[0004] BTK is a member of the non-receptor tyrosine kinase TEC family, and its structure includes PH domain, TH domain, SH3 domain, SH2 domain and SH1 domain. BTK plays a key role in the activation of the B cell antigen receptor (BCR) signaling pathway, regulates the development and activation of B cells, and plays an important role in the proliferation of B cells, the expression of proinflammatory cytokines and the secretion of antibodies (Targeting Bruton's tyrosine kinase in B cell malignancies. Nat Rev Cancer. 2014 Apr; 14(4): 219-32). Therefore, BTK has become one of the important targets for the treatment of diseases related to abnormal B cell activation, including autoimmune diseases and B cell lymphoma. Ibrutinib, Acalabrutinib and Zanubrutinib are three approved BTK inhibitors, which are mainly used to treat B cell lymphoma. They have obvious efficacy in some patients, but serious side effects and drug-resistant mutations have also been observed in clinical practice. In 2017, ibrutinib was approved by the US FDA for the treatment of graft-versus-host disease (GVHD), and other BTK inhibitors are currently being actively explored in the clinic for the treatment of autoimmune diseases, including RA, SLE, and multiple sclerosis (MS).

[0005] JAK3 is a member of the JAK family of non-receptor tyrosine kinases. The JAK kinase family has four members: JAK-1, JAK-2, JAK-3, and TYK-2. Signal transducers and activators of transcription (STAT) are downstream substrates of JAK3. JAK3 activates STAT to form dimers and enter the cell nucleus to regulate the transcriptional expression of specific genes. The JAK-STAT signaling pathway plays an important role in lymphocyte proliferation, differentiation, and the expression of proinflammatory cytokines (JAK inhibition as a therapeutic strategy for immune and inflammatory diseases. Nat Rev Drug Discov. 2017 December 28; 17(1): 78; The JAK-STAT Pathway: Impact on Human Disease and Therapeutic Intervention. Annual Review of Medicine. Vol. 66: 311-328), so JAK3 has become one of the targets of autoimmune diseases and malignant tumors. Tofacitinib is an FDA-approved JAK3 inhibitor that has shown good clinical efficacy in RA and IBD. However, there are certain adverse reactions, including severe infection and liver damage, which are believed to be related to the insufficient selectivity of Tofacitinib for JAK1 / 2 (JAK inhibition as a therapeutic strategy for immune and inflammatory diseases. Nat Rev Drug Discov. 2017 December 28; 17(1): 78; JAK-inhibitors. New players in the field of immune-mediated diseases, beyond rheumatoid arthritis. Rheumatology (Oxford). 2019 Feb 1; 58(Suppl 1): i43-i54).

[0006] In addition to the individual clinical effects of BTK and JAK3 inhibitors, simultaneous inhibition of the BTK / JAK3 signaling pathway will show synergistic therapeutic effects. Several studies have shown that simultaneous inhibition of BTK and JAK in the collagen-induced rat arthritis model (CIA) showed significant relief of joint swelling, a decrease in the number of osteoclasts, and a significant improvement in pathological scores, with efficacy superior to that of single drugs (2016 ACR / ARHP Annual Meeting.Abstract 484; 2013 ACR / ARHP AnnualMeeting.Abstract 2353). Abbvie launched Phase II clinical trials of ABBV599 (combination of BTK inhibitor and JAK inhibitor) for RA and SLE in September 2018 and June 2019, respectively. Another BTK / JAK3 dual-target inhibitor, DWP212525, also showed disease relief and joint protection in the mouse CIA model (2019 ACR / ARHP Annual Meeting.Abstract 965).

[0007] In view of the huge autoimmune disease market and unmet market demand, based on the functions of BTK and JAK3 in autoimmune diseases and existing clinical effects, it is necessary to develop dual-target small molecule inhibitors against BTK and JAK3 with good activity, good selectivity and low toxic side effects. Summary of the invention

[0008] The present invention provides a compound represented by formula (Ib) or a pharmaceutically acceptable salt thereof:

[0009]

[0010] in:

[0011] X is selected from O or NH;

[0012] R 1 Selected from H or optionally R a Substituted with the following groups: C1-C 10 Alkyl, C3-C 14 Cycloalkyl, 3-14 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl;

[0013] R a Selected from F, Cl, Br, I, OH, CN, =O, NO2 or optionally R b Substituted with the following groups: NH2, SH, S(O)NH2, S(O)(C1-C 10 Alkyl), S(O)2(C1-C 10 Alkyl), P(O)(C1-C10 Alkyl), C1-C 10 Alkyl, C3-C 14 Cycloalkyl, 3-14 membered heterocyclic group, C1-C 10 Alkoxy, C3-C 14 Cycloalkyloxy, 3-14 membered heterocyclyloxy, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C6-C 10 Aryl, 5-10 membered heteroaryl, C6-C 10 Aryloxy or 5-10 membered heteroaryloxy;

[0014] R b Selected from F, Cl, Br, I, OH, CN, =O, NO2, NH2, SH, C1-C 10 Alkyl, C3-C 14 Cycloalkyl, 3-14 membered heterocyclic group, C1-C 10 Alkoxy, C3-C 14 Cycloalkyloxy, 3-14 membered heterocyclyloxy, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C6-C 10 Aryl, 5-10 membered heteroaryl, C6-C 10 Aryloxy or 5-10 membered heteroaryloxy;

[0015] R 2 Selected from H, F, Cl, Br, I, C3-C 14 Cycloalkyl or phenyl, the C3-C 14 Cycloalkyl or phenyl is optionally replaced by R d replace;

[0016] R d C1-C4 alkyl selected from F, Cl, Br, I, OH, CN or optionally substituted by a group selected from F, Cl, Br, I, OH;

[0017] R 3 is selected from H, F, Cl, Br, I or a C1-C ... 10 alkyl;

[0018] R 4 is selected from H, F, Cl, Br, I, OH, CN or optionally R c Substituted with the following groups: C1-C 10 Alkyl, C1-C 10 Alkoxy;

[0019] R 5 , R 6 , R7 independently selected from H, F, Cl, Br, I, CN or optionally R e Substituted with the following groups: C1-C 10 Alkyl, C3-C 10 Cycloalkyl or 3-10 membered heterocyclic group;

[0020] R c , R e Independently selected from F, Cl, Br, I, OH;

[0021] n is selected from 0 or 1;

[0022] m is selected from 1 or 2;

[0023] The condition is: when R 2 =Cl, X = O, n = 1 and R 1 is not methyl or cyclopropyl; when R 2 =H, X = O and R 1 Not ethyl, CH2CHF2, CH2CH2OH, cyclopropyl, When R 2 =F, R 1 Not ethyl or CH2CHF2.

[0024] In some embodiments, X is selected from NH.

[0025] In some embodiments, X is selected from O.

[0026] In some embodiments, R 2 Selected from C3-C 14 Cycloalkyl or phenyl, the C3-C 14 Cycloalkyl or phenyl is optionally replaced by R d replace.

[0027] In some embodiments, R 2 Selected from C3-C 10 Cycloalkyl or phenyl, the C3-C 10 Cycloalkyl or phenyl is optionally replaced by R d replace.

[0028] In some embodiments, R 2 is selected from C3-C6 cycloalkyl or phenyl, wherein the C3-C6 cycloalkyl or phenyl is optionally replaced by R d replace.

[0029] In some embodiments, R 2 is selected from cyclopropyl or phenyl, wherein the cyclopropyl or phenyl is optionally replaced by R d replace.

[0030] In some embodiments, R 2Selected from C3-C6 cycloalkyl or phenyl.

[0031] In some embodiments, R 2 Selected from cyclopropyl or phenyl.

[0032] In some embodiments, R 2 is selected from H, F, Cl, Br or I.

[0033] In some embodiments, R 2 is selected from H, F or Cl.

[0034] In some embodiments, R 3 C1-C6 alkyl is selected from H, F, Cl, Br, I or optionally substituted by a group selected from F, Cl, Br, I, OH.

[0035] In some embodiments, R 3 is selected from H, F, Cl, Br, I or a C1-C4 alkyl group optionally substituted by a group selected from F, Cl, Br, I, OH.

[0036] In some embodiments, R 3 is selected from H, F, Cl, Br or I.

[0037] In some embodiments, R 3 Select from H or F.

[0038] In some embodiments, R 1 Selected from H or optionally R a Substituted with the following groups: C1-C 10 Alkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl.

[0039] In some embodiments, R 1 Selected from C1-C 10 Alkyl or 3-10 membered heterocyclic group, the C1-C 10 The alkyl group or the 3-10 membered heterocyclic group is optionally replaced by R a replace.

[0040] In some embodiments, R 1 is selected from C1-C6 alkyl or 4-6 membered heterocyclic group, wherein the C1-C6 alkyl or 4-6 membered heterocyclic group is optionally replaced by R a replace.

[0041] In some embodiments, R 1 is selected from C1-C6 alkyl or 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group contains O atoms and / or N atoms, and the C1-C6 alkyl or 4-6 membered heterocyclic group is optionally replaced by Ra replace.

[0042] In some embodiments, R 1 is selected from C1-C6 alkyl or 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group contains O and / or N as ring atoms, and the C1-C6 alkyl or 4-6 membered heterocyclic group is optionally replaced by R a replace.

[0043] In some embodiments, R 1 is selected from C1-C6 alkyl or 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group contains one O atom or one N atom, and the C1-C6 alkyl or 4-6 membered heterocyclic group is optionally replaced by R a replace.

[0044] In some embodiments, R 1 is selected from C1-C6 alkyl or 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group contains one O or one N as a ring atom, and the C1-C6 alkyl or 4-6 membered heterocyclic group is optionally replaced by R a replace.

[0045] In some embodiments, R a Selected from F, Cl, Br, I, OH, CN, =O, NO2 or optionally R b Substituted with the following groups: NH2, SH, C1-C 10 Alkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C1-C 10 Alkoxy, C3-C 10 Cycloalkyloxy, 3-10 membered heterocyclyloxy, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C6-C 10 Aryl, 5-10 membered heteroaryl, C6-C 10 aryloxy or 5-10 membered heteroaryloxy.

[0046] In some embodiments, R b Selected from F, Cl, Br, I, OH, CN, =O, NO2, NH2, SH, C1-C 10 Alkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C1-C 10 Alkoxy, C3-C 10 Cycloalkyloxy, 3-10 membered heterocyclyloxy, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C6-C 10 Aryl, 5-10 membered heteroaryl, C6-C 10 aryloxy or 5-10 membered heteroaryloxy.

[0047] In some embodiments, R a Selected from F, Cl, Br, I, OH, CN, C1-C 10 Alkyl or C3-C 10 Cycloalkyl.

[0048] In some embodiments, R a Selected from F, Cl, Br, I or C1-C6 alkyl.

[0049] In some embodiments, R a Selected from F or C1-C6 alkyl.

[0050] In some embodiments, R a Selected from C1-C6 alkyl.

[0051] In some embodiments, R a Selected from F or methyl.

[0052] In some embodiments, R a Selected from F.

[0053] In some embodiments, R a Selected from methyl.

[0054] In some embodiments, R 1 is selected from methyl, trifluoromethyl, ethyl, oxetanyl, tetrahydrofuranyl, N-methylpyrrolidinyl, tetrahydropyranyl or N-methylpiperidinyl.

[0055] In some embodiments, R 1 is selected from methyl, ethyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl or N-methylpiperidinyl.

[0056] In some embodiments, R 1 Selected from ethyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl or N-methylpiperidinyl.

[0057] In some embodiments, R 1 Selected from methyl, trifluoromethyl, ethyl,

[0058] In some embodiments, R 1 Selected from methyl, ethyl,

[0059] In some embodiments, R 1 Selected from ethyl,

[0060] In some embodiments, R 4is selected from H, F, Cl, Br, I, OH, CN or optionally R c Substituted with the following groups: C1-C 10 alkyl.

[0061] In some embodiments, R 4 Selected from H, F, Cl, Br, I, OH, CN or C1-C6 alkyl.

[0062] In some embodiments, R 4 Selected from H or C1-C6 alkyl.

[0063] In some embodiments, R 4 Selected from H or methyl.

[0064] In some embodiments, R 5 is selected from H, F, Cl, Br, I, CN or optionally R e Substituted C1-C 10 alkyl.

[0065] In some embodiments, R 5 Selected from H, F, Cl, Br, I, CN.

[0066] In some embodiments, R 5 Selected from H or CN.

[0067] In some embodiments, R 6 , R 7 independently selected from H, F, Cl, Br, I, CN or optionally R e Substituted C1-C 10 alkyl.

[0068] In some embodiments, R 6 , R 7 independently selected from H, CN or optionally R e Substituted C1-C 10 alkyl.

[0069] In some embodiments, R 6 , R 7 Independently selected from H, CN or C1-C6 alkyl.

[0070] In some embodiments, R 6 , R 7 Independently selected from H or C1-C6 alkyl.

[0071] In some embodiments, R 6 , R 7 are independently selected from H or tert-butyl.

[0072] In some embodiments, n is selected from 1.

[0073] In some embodiments, n is selected from 0.

[0074] In some embodiments, said m is selected from 1.

[0075] In some embodiments, m is selected from 2.

[0076] In some embodiments, the compound of formula (Ib) or a pharmaceutically acceptable salt thereof is selected from a compound of formula (Ia) or a pharmaceutically acceptable salt thereof:

[0077]

[0078] in:

[0079] X is selected from O or NH;

[0080] R 1 Selected from H or optionally R a Substituted with the following groups: C1-C 10 Alkyl, C3-C 14 Cycloalkyl, 3-14 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl;

[0081] R a Selected from F, Cl, Br, I, OH, CN, =O, NO2 or optionally R b Substituted with the following groups: NH2, SH, S(O)NH2, S(O)(C1-C 10 Alkyl), S(O)2(C1-C 10 Alkyl), P(O)(C1-C 10 Alkyl), C1-C 10 Alkyl, C3-C 14 Cycloalkyl, 3-14 membered heterocyclic group, C1-C 10 Alkoxy, C3-C 14 Cycloalkyloxy, 3-14 membered heterocyclyloxy, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C6-C 10 Aryl, 5-10 membered heteroaryl, C6-C 10 Aryloxy or 5-10 membered heteroaryloxy;

[0082] R b Selected from F, Cl, Br, I, OH, CN, =O, NO2, NH2, SH, C1-C 10 Alkyl, C3-C 14 Cycloalkyl, 3-14 membered heterocyclic group, C1-C 10 Alkoxy, C3-C 14Cycloalkyloxy, 3-14 membered heterocyclyloxy, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C6-C 10 Aryl, 5-10 membered heteroaryl, C6-C 10 Aryloxy or 5-10 membered heteroaryloxy;

[0083] R 2 Selected from C3-C 14 Cycloalkyl or phenyl, the C3-C 14 Cycloalkyl or phenyl is optionally replaced by R d replace;

[0084] R d C1-C4 alkyl selected from F, Cl, Br, I, OH, CN or optionally substituted by a group selected from F, Cl, Br, I, OH;

[0085] R 3 is selected from H, F, Cl, Br, I or a C1-C ... 10 alkyl;

[0086] R 4 is selected from H, F, Cl, Br, I, OH, CN or optionally R c Substituted with the following groups: C1-C 10 Alkyl, C1-C 10 Alkoxy;

[0087] R c Selected from F, Cl, Br, I, OH;

[0088] n is selected from 0 or 1.

[0089] In some embodiments, X is selected from NH.

[0090] In some embodiments, R 2 Selected from C3-C 10 Cycloalkyl or phenyl, the C3-C 10 Cycloalkyl or phenyl is optionally replaced by R d replace.

[0091] In some embodiments, R 2 is selected from C3-C6 cycloalkyl or phenyl, wherein the C3-C6 cycloalkyl or phenyl is optionally replaced by R d replace.

[0092] In some embodiments, R 2 is selected from cyclopropyl or phenyl, wherein the cyclopropyl or phenyl is optionally replaced by R d replace.

[0093] In some embodiments, R 2 Selected from C3-C6 cycloalkyl or phenyl.

[0094] In some embodiments, R 2 Selected from cyclopropyl or phenyl.

[0095] In some embodiments, R 3 C1-C6 alkyl is selected from H, F, Cl, Br, I or optionally substituted by a group selected from F, Cl, Br, I, OH.

[0096] In some embodiments, R 3 is selected from H, F, Cl, Br, I or a C1-C4 alkyl group optionally substituted by a group selected from F, Cl, Br, I, OH.

[0097] In some embodiments, R 3 is selected from H, F, Cl, Br or I.

[0098] In some embodiments, R 3 Select from H or F.

[0099] In some embodiments, R 1 Selected from H or optionally R a Substituted with the following groups: C1-C 10 Alkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl.

[0100] In some embodiments, R 1 Selected from C1-C 10 Alkyl or 3-10 membered heterocyclic group, the C1-C 10 The alkyl group or the 3-10 membered heterocyclic group is optionally replaced by R a replace.

[0101] In some embodiments, R 1 is selected from C1-C6 alkyl or 4-6 membered heterocyclic group, wherein the C1-C6 alkyl or 4-6 membered heterocyclic group is optionally replaced by R a replace.

[0102] In some embodiments, R 1 is selected from C1-C6 alkyl or 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group contains O atoms and / or N atoms, and the C1-C6 alkyl or 4-6 membered heterocyclic group is optionally replaced by R a replace.

[0103] In some embodiments, R 1is selected from C1-C6 alkyl or 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group contains O and / or N as ring atoms, and the C1-C6 alkyl or 4-6 membered heterocyclic group is optionally replaced by R a replace.

[0104] In some embodiments, R 1 is selected from C1-C6 alkyl or 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group contains one O atom or one N atom, and the C1-C6 alkyl or 4-6 membered heterocyclic group is optionally replaced by R a replace.

[0105] In some embodiments, R 1 is selected from C1-C6 alkyl or 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group contains one O or one N as a ring atom, and the C1-C6 alkyl or 4-6 membered heterocyclic group is optionally replaced by R a replace.

[0106] In some embodiments, R a Selected from F, Cl, Br, I, OH, CN, =O, NO2 or optionally R b Substituted with the following groups: NH2, SH, C1-C 10 Alkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C1-C 10 Alkoxy, C3-C 10 Cycloalkyloxy, 3-10 membered heterocyclyloxy, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C6-C 10 Aryl, 5-10 membered heteroaryl, C6-C 10 aryloxy or 5-10 membered heteroaryloxy.

[0107] In some embodiments, R b Selected from F, Cl, Br, I, OH, CN, =O, NO2, NH2, SH, C1-C 10 Alkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C1-C 10 Alkoxy, C3-C 10 Cycloalkyloxy, 3-10 membered heterocyclyloxy, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C6-C 10 Aryl, 5-10 membered heteroaryl, C6-C 10 aryloxy or 5-10 membered heteroaryloxy.

[0108] In some embodiments, R aSelected from F, Cl, Br, I, OH, CN, C1-C 10 Alkyl or C3-C 10 Cycloalkyl.

[0109] In some embodiments, R a Selected from C1-C6 alkyl.

[0110] In some embodiments, R a In some embodiments, R 1 is selected from methyl, ethyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl or N-methylpiperidinyl.

[0111] In some embodiments, R 1 Selected from ethyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl or N-methylpiperidinyl.

[0112] In some embodiments, R 1 Selected from methyl, ethyl,

[0113] In some embodiments, R 1 Selected from ethyl,

[0114] In some embodiments, R 4 is selected from H, F, Cl, Br, I, OH, CN or optionally R c Substituted with the following groups: C1-C 10 alkyl.

[0115] In some embodiments, R 4 Selected from H, F, Cl, Br, I, OH, CN or C1-C6 alkyl.

[0116] In some embodiments, R 4 Selected from H or C1-C6 alkyl.

[0117] In some embodiments, R 4 Selected from H or methyl.

[0118] In some embodiments, n is selected from 1.

[0119] In some embodiments, n is selected from 0. In some embodiments, the compound of formula (Ib) or a pharmaceutically acceptable salt thereof is selected from a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0120]

[0121] Where: R 1 , R 2 , R 3As defined in formula (Ib).

[0122] In some embodiments, the compound of formula (Ib) or a pharmaceutically acceptable salt thereof is selected from a compound of formula (IIb) or a pharmaceutically acceptable salt thereof:

[0123]

[0124] Where: R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , X, m and n are as defined in formula (Ib).

[0125] In some embodiments, the compound of formula (Ib) or a pharmaceutically acceptable salt thereof is selected from a compound of formula (IIa) or a pharmaceutically acceptable salt thereof:

[0126]

[0127] Among them, X, R 1 , R 2 , R 3 , R 4 and n are as defined above.

[0128] In some embodiments, the compound of formula (Ib) or a pharmaceutically acceptable salt thereof is selected from a compound of formula (II) or a pharmaceutically acceptable salt thereof:

[0129]

[0130] Among them, R 1 , R 2 , R 3 As defined above.

[0131] In some embodiments, the compound represented by formula (Ib) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0132]

[0133] In some embodiments, the compound represented by formula (Ib) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts:

[0134]

[0135] The present invention also provides a pharmaceutical composition comprising a compound represented by formula (Ib) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0136] The present invention also provides a pharmaceutical composition comprising a compound represented by formula (Ib) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient.

[0137] Furthermore, the present invention relates to the use of a compound represented by formula (Ib) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a drug for preventing or treating Janus kinase (JAK, especially JAK3) and / or Bruton's tyrosine kinase (BTK) related diseases.

[0138] Furthermore, the present invention relates to the use of a compound represented by formula (Ib) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in preventing or treating diseases related to Janus kinase (JAK, especially JAK3) and / or Bruton's tyrosine kinase (BTK).

[0139] Furthermore, the present invention relates to a compound of formula (Ib) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for preventing or treating diseases related to Janus kinase (JAK, especially JAK3) and / or Bruton's tyrosine kinase (BTK).

[0140] The present invention also relates to a method for treating Janus kinase (JAK, especially JAK3) and / or Bruton's tyrosine kinase (BTK) related diseases, which comprises administering to a patient a therapeutically effective dose of a pharmaceutical preparation comprising a compound of formula (Ib) or a pharmaceutically acceptable salt thereof according to the present invention.

[0141] In a preferred embodiment of the present invention, the Janus kinase (JAK, especially JAK3) and / or Bruton's tyrosine kinase (BTK) related diseases include but are not limited to tumors (such as B cell lymphoma) and autoimmune diseases (such as rheumatoid arthritis, inflammatory bowel inflammation and systemic lupus erythematosus), etc.

[0142] Definitions and explanations of terms

[0143] Unless otherwise specified, the definitions of groups and terms recorded in the specification and claims of the present invention, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be arbitrarily combined and coupled with each other. The group definitions and compound structures after such combinations and couplings shall fall within the scope of the description of the present invention.

[0144] In the present invention The ligation sites are indicated.

[0145] The term "pharmaceutically acceptable salt" refers to salts of pharmaceutically acceptable non-toxic acids or bases, including salts of inorganic acids and bases, and organic acids and bases.

[0146] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, diastereomers and conformational isomers.

[0147] The compounds of the present invention may have asymmetric carbon atoms (optical centers) or double bonds. Racemates, enantiomers, diastereomers, geometric isomers and individual isomers are all included within the scope of the present invention.

[0148] The graphic representations of racemates or enantiomerically pure compounds herein are from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise indicated, the wedge and dashed wedge keys are used. To indicate the absolute configuration of a stereocenter, use black real and imaginary bonds. represents the cis-trans configuration of alicyclic compounds. When the compounds described herein contain olefinic double bonds or other geometric asymmetric centers, unless specified otherwise, they include E and Z geometric isomers. Likewise, all tautomeric forms are included within the scope of the present invention.

[0149] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as mixtures enriched in enantiomers or diastereomers, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present invention. The compounds containing asymmetric atoms of the present application can be isolated in optically pure form or in racemic form. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral raw materials or chiral reagents.

[0150] The term "tautomer" refers to functional group isomers resulting from the rapid movement of an atom in a molecule between two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Prototropic tautomers arise from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to separate a single tautomer usually produce a mixture whose physicochemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical characteristics within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; while in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.

[0151] The term "pharmaceutical composition" refers to a mixture of one or more compounds described herein or their physiologically / pharmaceutically acceptable salts or prodrugs with other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.

[0152] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are replaced, and oxo will not occur on an aromatic group.

[0153] The term "optional" or "optionally" means that the event or situation described subsequently may or may not occur, and the description includes the occurrence of the event or situation and the non-occurrence of the event or situation. For example, an ethyl group is "optionally" substituted with a halogen, meaning that the ethyl group may be unsubstituted (CH2CH3), monosubstituted (such as CH2CH2F), polysubstituted (such as CHFCH2F, CH2CHF2, etc.), or fully substituted (CF2CF3). It will be appreciated by those skilled in the art that for any group containing one or more substituents, no substitution or substitution pattern that is sterically impossible to exist and / or cannot be synthesized will be introduced.

[0154] When any variable (eg, R) occurs more than one time in a compound composition or structure, its definition at each occurrence is independent.

[0155] The term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

[0156] The term "C1-C 10 “Alkyl” is understood as meaning a straight-chain or branched, saturated, monovalent hydrocarbon radical having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. The alkyl radical is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc.; "C1-C6 alkyl" should be understood to mean a straight chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5, or 6 carbon atoms. "C1-C4 alkyl" should be understood to mean a straight chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, or 4 carbon atoms. The term "alkoxy" can be understood as "alkyloxy" or "alkyl-O", preferably, "C1-C 10The "alkoxy group" may include a "C1-C6 alkoxy group" and a "C1-C4 alkoxy group".

[0157] The term "C2-C 10 "Alkenyl" is understood to mean preferably a linear or branched monovalent hydrocarbon group containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9, 10 carbon atoms, preferably "C2-C6 alkenyl", further preferably "C2-C4 alkenyl", and further preferably C2 or C3 alkenyl. It should be understood that in the case where the alkenyl contains more than one double bond, the double bonds may be separated from each other or conjugated. The alkenyl group is, for example, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl, (Z)-1-methylprop-1-enyl.

[0158] The term "C2-C 10 "Alkynyl" is understood to mean a linear or branched monovalent hydrocarbon radical containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9, 10 carbon atoms, preferably "C2-C6 alkynyl", further preferably "C2-C4 alkynyl", further preferably C2 or C3 alkynyl. The alkynyl radical is, for example, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, 1-methylprop-2-ynyl.

[0159] The term "C3-C 14 "Cycloalkyl" is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 14 carbon atoms. 10 "Cycloalkyl" is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 10 carbon atoms. The term "C3-C6 cycloalkyl" is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3, 4, 5 or 6 carbon atoms. Examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon ring such as a decalin ring. According to the present invention, the bicyclic hydrocarbon ring includes a bridged ring, a spiro ring or a paracyclic ring structure.

[0160] The term "cycloalkyloxy" may be understood as "cycloalkyl-O", preferably, "C3-C 14 The cycloalkyloxy group may contain a C3-C 10 "Cycloalkyloxy" and "C3-C6 cycloalkyloxy".

[0161] The term "3-14 membered heterocyclyl" is understood as a saturated or partially saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3-14 ring atoms, which contains 1-5, preferably 1-3 heteroatoms selected from N, O and S. The term "3-10 membered heterocyclyl" means a saturated or partially saturated monovalent monocyclic or bicyclic hydrocarbon ring containing 1-5, preferably 1-3 heteroatoms selected from N, O and S. The term "4-6 membered heterocyclyl" is understood as a saturated or partially saturated monovalent monocyclic or bicyclic hydrocarbon ring having 4, 5, 6 ring atoms, which contains 1-5, preferably 1-3 heteroatoms selected from N, O and S. In particular, the heterocyclic group may include, but is not limited to, a 4-membered ring, such as azetidinyl, oxetanyl; a 5-membered ring, such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or a partially saturated 6-membered ring such as tetrahydropyridinyl; or a 7-membered ring, such as diazepanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclic group may be bicyclic, such as, but not limited to, a 5,5-membered ring, such as a hexahydrocyclopenta [c] pyrrole-2 (1H) -yl ring, or a 5,6-membered bicyclic ring, such as a hexahydropyrrolo [1,2-a] pyrazine-2 (1H) -yl ring. The ring containing the nitrogen atom may be partially unsaturated, i.e. it may contain one or more double bonds, such as but not limited to 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl, 4,5-dihydrooxazolyl or 4H-[1,4]thiazinyl, or it may be benzo-fused, such as but not limited to dihydroisoquinolinyl. According to the present invention, the heterocyclic group is non-aromatic. The bicyclic hydrocarbon ring includes a bridged ring, a spiro ring or a cyclic structure.

[0162] The term “3-14-membered heterocyclyloxy” may be understood as “3-14-membered heterocyclyl-O”, and preferably, “3-14-membered heterocyclyloxy” may include “3-10-membered heterocyclyloxy”.

[0163] The term "C6-C 10 The term "aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic or bicyclic hydrocarbon ring having 6, 7, 8, 9 or 10 carbon atoms. In particular, a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9 aryl"), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C 10 "aryl"), for example tetrahydronaphthyl, dihydronaphthyl or naphthyl.

[0164] The term "C6-C 10 "Aryloxy" can be understood as "C6-C 10 Aryl-O".

[0165] “5-10 membered heteroaryl” is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10 ring atoms, in particular 5 or 6 or 9 or 10 ring atoms, and which contain 1 to 5, preferably 1 to 3 heteroatoms independently selected from N, O and S and, in addition, in each case may be benzo-fused. In particular, the heteroaryl group is selected from thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl and the like and benzo derivatives thereof, such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl and the like and benzo derivatives thereof, such as quinolyl, quinazolinyl, isoquinolyl and the like; or azinyl, indolizinyl, purinyl and the like and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl and the like.

[0166] The term "5-10 membered heteroaryloxy" may be understood as "5-10 membered heteroaryl-O".

[0167] The present application also includes isotopically labeled compounds of the present application that are identical to those described herein, but in which one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 Cl et al.

[0168] Certain isotope-labeled compounds of the present application (e.g. 3 H and 14 C-labeled) can be used in compound and / or substrate tissue distribution assays. 3 H) and carbon-14 (i.e. 14 C) isotopes are particularly preferred due to their ease of preparation and detectability. Positron emitting isotopes such as 15 O.13 N. 11 C and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the present application can generally be prepared by following procedures analogous to those disclosed in the Schemes and / or Examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0169] In addition, the use of heavier isotopes such as deuterium (i.e. 2 H)) substitution may offer certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and may therefore be preferred in certain circumstances, wherein deuterium substitution may be partial or complete, partial deuterium substitution meaning that at least one hydrogen is replaced by at least one deuterium.

[0170] All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.

[0171] The term "treatment" means administering the compound or formulation described herein to prevent, improve or eliminate a disease or one or more symptoms associated with the disease, and includes:

[0172] (i) preventing a disease or disease state from occurring in a mammal, particularly where such mammal is susceptible to such disease state but has not yet been diagnosed as having such disease state;

[0173] (ii) inhibiting a disease or disease state, i.e., arresting its development;

[0174] (iii) ameliorating a disease or condition, even if causing regression of the disease or condition.

[0175] The term "therapeutically effective amount" means an amount of a compound of the present invention that (i) treats or prevents a specific disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a specific disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a specific disease, condition, or disorder described herein. The amount of a compound of the present invention that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by those skilled in the art based on their own knowledge and this disclosure.

[0176] The term "excipient" refers to a pharmaceutically acceptable inert ingredient. Examples of the term "excipient" include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, and diluents. Excipients can enhance the handling characteristics of a pharmaceutical preparation, i.e., make the preparation more suitable for direct compression by increasing fluidity and / or adhesion. Examples of typical "pharmaceutically acceptable carriers" suitable for the above-mentioned preparations are: sugars, starches, cellulose and its derivatives, etc., which are commonly used excipients in pharmaceutical preparations.

[0177] The term "pharmaceutically acceptable excipients" refers to those excipients that have no significant irritation to the organism and do not impair the biological activity and performance of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

[0178] The words “comprise”, “comprise” or “comprises” and their English variations such as comprises or comprising should be construed in an open, non-exclusive sense, ie, “including but not limited to”.

[0179] The pharmaceutical composition of the present application can be prepared by combining the compound of the present application with suitable pharmaceutically acceptable excipients, for example, it can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols, etc.

[0180] Typical routes of administration of the compounds of the present application or their pharmaceutically acceptable salts or their pharmaceutical compositions include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0181] The pharmaceutical composition of the present application can be manufactured by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, sugar-coated pill making methods, grinding methods, emulsification methods, freeze-drying methods, etc.

[0182] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be prepared by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present application to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, etc., for oral administration to patients.

[0183] Solid oral compositions can be prepared by conventional mixing, filling or tableting methods. For example, they can be obtained by mixing the active compound with a solid excipient, optionally grinding the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into particles to obtain a tablet or dragee core. Suitable excipients include, but are not limited to, adhesives, diluents, disintegrants, lubricants, glidants, sweeteners or flavoring agents, etc.

[0184] The pharmaceutical composition may also be suitable for parenteral administration, such as sterile solutions, suspensions or lyophilized products in appropriate unit dosage forms.

[0185] In all administration methods of the compounds of general formula (Ib) described herein, the daily dosage is 0.01 to 100 mg / kg body weight, preferably 0.05 to 50 mg / kg body weight, more preferably 0.1 to 30 mg / kg body weight, in the form of single or divided doses. The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitution methods well known to those skilled in the art, and preferred embodiments include but are not limited to the embodiments of the present invention.

[0186] The chemical reactions of the specific embodiments of the present invention are carried out in a suitable solvent, which must be suitable for the chemical changes of the present invention and the reagents and materials required. In order to obtain the compounds of the present invention, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction processes based on the existing embodiments. DETAILED DESCRIPTION

[0187] The following examples illustrate the technical solutions of the invention in detail, but the protection scope of the present invention includes but is not limited to them.

[0188] The solvents used in the present invention are commercially available. Commercially available compounds are referred to by their supplier catalog names.

[0189] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The units of NMR shifts are 10 -6 (ppm). The solvents for NMR determination are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is tetramethylsilane (TMS); "IC 50 " refers to the half inhibition concentration, which refers to the concentration when the maximum inhibition effect is half achieved. The eluent below can be formed by two or more solvents to form a mixed eluent, and the percentage is the volume ratio of each solvent. For example, "methanol / dichloromethane: 0-8%" means that during the gradient elution process, the volume ratio of methanol:dichloromethane in the mixed eluent is 0:100 to 8:100.

[0190] The present invention uses the following abbreviations:

[0191]

[0192] Example 1 Preparation of Compound 001

[0193] (R)-1-(3-((5-cyclopropyl-2-((1-ethyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one

[0194]

[0195] Synthesis route and specific synthesis steps:

[0196]

[0197] Step 1: Synthesis of 2,4-dichloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (1b)

[0198] 2,4-Dichloro-7H-pyrrolo[2,3-d]pyrimidine 1a (10 g, 53.2 mmol), N-iodosuccinimide (14.4 g, 79.8 mmol) and 2,4,6-trimethylaniline (718 mg, 0.53 mmol) were dissolved in dichloromethane (100 mL) at room temperature, and the reaction solution was stirred at 110°C for 16 hours until the reaction was complete. The mixture was poured into water and extracted with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column purification (EA:PE = 0-20%) to obtain the title product 1b (13.0 g).

[0199] LCMS:Rt:1.708min; MS m / z(ESI):313.8[M+H] + .

[0200] Step 2: Synthesis of 2,4-dichloro-5-iodo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine (1c)

[0201] 2,4-Dichloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine 1b (7 g, 22.4 mmol) was dissolved in tetrahydrofuran (70 mL) at room temperature, sodium hydride (3.58 g, 89.6 mmol, 60%) and 2-(trimethylsilyl)ethoxymethyl chloride (5.6 g, 33.6 mmol) were added in sequence at 0°C, and the reaction solution was stirred at 0°C for 0.5 hours until the reaction was complete. 50 mL of water was slowly added to the reaction solution to quench the reaction, the mixture was poured into water and extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column purification (EA:PE = 0-20%) to obtain the title product 1c (3.5 g).

[0202] LCMS:Rt:1.230min;MS m / z(ESI):444.0[M+H] + .

[0203] Step 3: Synthesis of 2,4-dichloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine (1e)

[0204] 2,4-Dichloro-5-iodo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine 1c (1.1 g, 2.5 mmol), cyclopropylboronic acid 1d (341 mg, 2.8 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (176 mg, 0.25 mmol), silver oxide (286 mg, 1.25 mmol) and potassium phosphate (1.6 g, 7.5 mmol) were added to a dioxane (12 mL) solution at room temperature, and then stirred at 90°C for 2 hours under a nitrogen atmosphere until the reaction was complete. The mixture was filtered, the liquid phase was poured into water and extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column purification (EA:PE = 0-20%) to obtain the title product 1e (500 mg).

[0205] LCMS:Rt:1.983min;MS m / z(ESI):358.0[M+H] + .

[0206] Step 4: Synthesis of tert-butyl (R)-3-((2-chloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate (1 g)

[0207] 2,4-Dichloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine 1e (480 mg, 1.3 mmol), (R)-3-aminopiperidine-1-carboxylic acid tert-butyl ester 1f (528 mg, 2.6 mmol) and N,N-diisopropylethylamine (521 mg, 3.9 mmol) were dissolved in isopropanol (5 mL) at room temperature, and the reaction solution was placed in a sealed tube and stirred at 110°C for 16 hours until the reaction was complete. The reaction solution was poured into water and extracted with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column purification (EA:PE = 0-20%) to obtain the title product 1g (580 mg).

[0208] LCMS:Rt:2.251min; MS m / z(ESI):522.2[M+H] + .

[0209] Step 5: Synthesis of tert-butyl (R)-3-((2-((1-ethyl-1H-pyrazol-4-yl)amino)-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate (1i)

[0210] Tert-butyl (R)-3-((2-chloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate 1g (560mg, 1.1mmol), 1-ethyl-1H-pyrazol-4-amine 1h (238mg, 2.2mmol) and trifluoroacetic acid (123mg, 1.1mmol) were dissolved in isopropanol (6mL) at room temperature, and the reaction solution was placed in a sealed tube and stirred at 110°C for 16 hours until the reaction was complete. The reaction solution was adjusted to pH = 9 with a saturated solution of NaHCO3, poured into water and extracted with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column purification (EA:PE = 0-10%) to obtain the title product 1i (100mg).

[0211] LCMS:Rt:1.440min;MS m / z(ESI):597.3[M+H] + .

[0212] Step 6: (R)-5-cyclopropyl-N 2 -(1-ethyl-1H-pyrazol-4-yl)-N 4 Synthesis of -(piperidin-3-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine-2,4-diamine (1j)

[0213] Tert-butyl (R)-3-((2-((1-ethyl-1H-pyrazol-4-yl)amino)-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate 1i (100 mg, 0.2 mmol) was dissolved in dichloromethane (1 mL) and dioxane hydrochloride (1 mL, 4 M) at room temperature, and the reaction solution was stirred at room temperature for 2 hours until the reaction was complete. The reaction solution was adjusted to pH = 9 with a saturated NaHCO3 solution, poured into water and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the title product 1j (80 mg, crude product).

[0214] LCMS:Rt:1.201min; MS m / z(ESI):497.3[M+H] + .

[0215] Step 7: Synthesis of (R)-1-(3-((5-cyclopropyl-2-((1-ethyl-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one (11)

[0216] At room temperature, (R)-5-cyclopropyl-N2-(1-ethyl-1H-pyrazol-4-yl)-N4-(piperidin-3-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine-2,4-diamine 1j (80 mg, 0.2 mmol) was dissolved in dichloromethane (2 mL), and triethylamine (64 mg, 0.6 mmol) and acryloyl chloride 1k (38 mg, 0.4 mmol) were added at -10°C. The reaction solution was continued to react at -10°C for 30 minutes until the reaction was complete. The reaction solution was diluted with clean water (20 mL), extracted with dichloromethane (20 mL*2), and the organic phases were combined and dried to obtain the title product 1l (100 mg).

[0217] LCMS:Rt:1.796min; MS m / z(ESI):551.3[M+H] + .

[0218] Step 8: Synthesis of (R)-1-(3-((5-cyclopropyl-2-((1-ethyl-1H-pyrazol-4-yl)amino)-7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one (1m)

[0219] (R)-1-(3-((5-cyclopropyl-2-((1-ethyl-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one 1l (100 mg, 0.2 mmol) and trifluoroacetic acid (103 mg, 0.9 mmol) were dissolved in dichloromethane (5 mL) at room temperature, and the reaction solution was stirred at room temperature for 0.5 hours until the reaction was complete. The reaction solution was adjusted to pH = 9 with a saturated NaHCO3 solution, poured into water and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the title product 1m (80 mg, crude product).

[0220] LCMS:Rt:1.003min; MS m / z(ESI):451.2[M+H] + .

[0221] Step 9: Synthesis of (R)-1-(3-((5-cyclopropyl-2-((1-ethyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one (001)

[0222] At room temperature, (R)-1-(3-((5-cyclopropyl-2-((1-ethyl-1H-pyrazol-4-yl)amino)-7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one 1m (80 mg, 0.2 mmol) was dissolved in tetrahydrofuran (3 mL), and sodium hydroxide (2 mL) solution was added. The reaction solution was stirred at room temperature for 1 hour until the reaction was complete. The reaction solution was diluted with water (20 mL), extracted with ethyl acetate (30 mL*2), and the combined organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the crude product was filtered and dried. The title product 001 (15.82 mg) was purified by high performance liquid preparative chromatography.

[0223] Preparative high performance liquid chromatography: Waters 2767 preparative chromatograph, mobile phase A: 10 mmol% FA in water, mobile phase B: acetonitrile, chromatographic column: Sunfire (PrepC18 OBD 19*250mm 10μm), gradient: 10% ACN for 4 min, 45%-55% ACN for 14 min, 95% ACN for 14 min, and 10% ACN for 17 min to complete purification and obtain the target compound.

[0224] LCMS:Rt:0.933min; MS m / z(ESI):421.2[M+H] + .

[0225] 1 H NMR (400MHz, DMSO-d6) δ10.59(d,J=8.3Hz,1H),8.43(s,1H),7.85(d,J=13.2Hz,1H),7.43(d,J=22.1H z,1H),6.89(d,J=5.7Hz,1H),6.45(s,1H),6.07(dd,J=38.5,16.3Hz,1H),5.70(ddd,J=91.0,51.4,9.1 Hz,2H),4.30(s,1H),4.03(q,J=7.2Hz,3H),3.80(d,J=10.9Hz,1H),3.68(d,J=14.3Hz,2H),1.97(s,1H ),1.85(s,2H),1.73(s,1H),1.57(s,1H),1.39–1.28(m,3H),0.76(d,J=9.4Hz,2H),0.59–0.41(m,2H).

[0226] Example 2 Preparation of Compound 002

[0227] (R)-1-(3-((2-((1-ethyl-1H-pyrazol-4-yl)amino)-5-phenyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one

[0228]

[0229] Synthesis route and specific synthesis steps:

[0230]

[0231] Step 1: Synthesis of 2,4-dichloro-5-phenyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine (2c)

[0232] 2,4-dichloro-5-iodo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine 2a (1.7 g, 3.8 mmol), phenylboronic acid 2b (466 mg, 3.8 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (112 mg, 0.15 mmol) and sodium carbonate (1.2 g, 11.4 mmol) were added to a solution of dioxane (20 mL) and water (4 mL) at room temperature, and then stirred at 85°C for 2 hours under nitrogen until the reaction was complete. The mixture was filtered, the liquid phase was poured into water and extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column purification (EA:PE = 0-20%) to obtain the title product 2c (900 mg).

[0233] LCMS:Rt:1.994min;MS m / z(ESI):394.0[M+H] + .

[0234] Step 2: Synthesis of tert-butyl (R)-3-((2-chloro-5-phenyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate (2e)

[0235] 2,4-Dichloro-5-phenyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine 2c (900 mg, 2.3 mmol), (R)-3-aminopiperidine-1-carboxylic acid tert-butyl ester 2d (930 mg, 4.6 mmol) and N,N-diisopropylethylamine (900 mg, 6.9 mmol) were dissolved in isopropanol (10 mL) at room temperature, and the reaction solution was stirred at 110°C for 16 hours until the reaction was complete. The reaction solution was poured into water and extracted with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column purification (EA:PE = 0-20%) to obtain the title product 2e (230 mg).

[0236] LCMS:Rt:2.467min; MS m / z(ESI):558.2[M+H] + .

[0237] Step 3: Synthesis of tert-butyl (R)-3-((2-((1-ethyl-1H-pyrazol-4-yl)amino)-5-phenyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate (2 g)

[0238] Tert-butyl (R)-3-((2-chloro-5-phenyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate 2e (230 mg, 0.4 mmol), 1-ethyl-1H-pyrazol-4-amine 2f (89 mg, 0.8 mmol) and trifluoroacetic acid (46 mg, 0.4 mmol) were dissolved in isopropanol (3 mL) at room temperature, and the reaction solution was stirred at 110°C for 16 hours until the reaction was complete. The reaction solution was adjusted to pH = 9 with a saturated solution of NaHCO3, poured into water and extracted with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column purification (EA:PE = 0-10%) to obtain the title product 2g (130 mg).

[0239] LCMS:Rt:1.749min; MS m / z(ESI):633.3[M+H] + .

[0240] Step 4: Synthesis of (R)-(2-((1-ethyl-1H-pyrazol-4-yl)amino)-5-phenyl-4-(piperidin-3-ylamino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methanol (2h)

[0241] Tert-butyl (R)-3-((2-((1-ethyl-1H-pyrazol-4-yl)amino)-5-phenyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate 2g (130mg, 0.2mmol) was dissolved in dichloromethane (1mL) and dioxane hydrochloride (1mL) solution at room temperature, and the reaction solution was stirred at room temperature for 16 hours until the reaction was complete. The reaction solution was adjusted to pH=9 with a saturated solution of NaHCO3, poured into water and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the title product 2h (100mg, crude product).

[0242] LCMS:Rt:0.682min;MS m / z(ESI):433.3[M+H] + .

[0243] Step 5: Synthesis of (R)-1-(3-((2-((1-ethyl-1H-pyrazol-4-yl)amino)-7-(hydroxymethyl)-5-phenyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one (2j)

[0244] (R)-(2-((1-ethyl-1H-pyrazol-4-yl)amino)-5-phenyl-4-(piperidin-3-ylamino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methanol 2h (90 mg, 0.2 mmol) was dissolved in dichloromethane (2 mL) at room temperature, and triethylamine (64 mg, 0.6 mmol) and acryloyl chloride 2i (38 mg, 0.4 mmol) were added at -10°C. The reaction solution was continued to react at -10°C for 30 minutes until the reaction was complete. The reaction solution was diluted with clean water (20 mL), extracted with dichloromethane (20 mL*2), and the organic phases were combined and dried to obtain the title product 2j (90 mg).

[0245] LCMS:Rt:1.253min;MS m / z(ESI):487.2[M+H] + .

[0246] Step 6: Synthesis of (R)-1-(3-((2-((1-ethyl-1H-pyrazol-4-yl)amino)-5-phenyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one (002)

[0247] At room temperature, (R)-1-(3-((2-((1-ethyl-1H-pyrazol-4-yl)amino)-7-(hydroxymethyl)-5-phenyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one 2j (90 mg, 0.2 mmol) was dissolved in tetrahydrofuran (3 mL), and NaOH (2 mL) solution was added. The reaction solution was stirred at room temperature for 30 minutes until the reaction was complete. The reaction solution was diluted with clean water (20 mL), extracted with ethyl acetate (30 mL*2), and the combined organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the crude product was filtered and dried. The title compound 002 (29.31 mg) was purified by high performance liquid preparative chromatography.

[0248] Preparative high performance liquid chromatography: Waters (waters) 2767 preparative chromatograph, mobile phase A (Mobile phase A): 10mmol% FA in water, mobile phase B (Mobile phase B): acetonitrile, chromatographic column: Sunfire (PrepC18 OBD 19*250mm 10μm), gradient (Gradient): 10% ACN run for 3min, 40%-45% ACN run to 13min, 95% ACN run to 15min, 10% ACN run to 19min to end purification and obtain the target compound.

[0249] LCMS:Rt:1.043min; MS m / z(ESI):457.2[M+H] + .

[0250] 1H NMR (400MHz, DMSO) δ11.19(s,1H),8.57(s,1H),7.88(d,J=9.7Hz,1H),7.44(dd,J=20.4,13 .3Hz,5H),7.30(dd,J=9.5,4.4Hz,1H),6.92–6.40(m,2H),6.03(dd,J=38.0,16.7Hz,1H),5 .58(dd,J=99.3,10.1Hz,1H),5.06(d,J=7.7Hz,1H),4.26(s,1H),4.05(q,J=6.9Hz,2H),3. 69(dd,J=35.0,14.2Hz,2H),3.52(s,2H),1.92(s,1H),1.49(s,2H),1.34(d,J=6.9Hz,4H).

[0251] Example 3 Preparation of Compound 003

[0252] 1-Acryloyl-(3S,4R)-3-((5-cyclopropyl-2-((1-ethyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-4-fluoropiperidine

[0253]

[0254] Synthesis route and specific synthesis steps:

[0255]

[0256] The preparation method was the same as that of Example 2, except that cyclopropylboronic acid (3b) was used instead of phenylboronic acid (2b) in Example 2, and (3S,4R)-3-amino-4-fluoropiperidine-1-carboxylic acid tert-butyl ester (3d) was used instead of (R)-3-aminopiperidine-1-carboxylic acid tert-butyl ester (2d) in Example 2, and the title compound 003 (8.84 mg) was purified by preparative HPLC.

[0257] Preparative high performance liquid chromatography: Waters 2767 preparative chromatograph, mobile phase A: 10 mmol% FA in water, mobile phase B: acetonitrile, chromatographic column: Sunfire (PrepC18 OBD 19*250mm 10μm), gradient: 10% ACN for 4 min, 45%-55% ACN for 14 min, 95% ACN for 14 min, and 10% ACN for 17 min to complete purification and obtain the target compound.

[0258] LCMS:Rt:0.869min;MS m / z(ESI):439.1[M+H] + .

[0259] 1 H NMR (400MHz, DMSO) δ10.68(s,1H),8.48(s,1H),7.94-7.78(m,1H),7.48-7.34(m,1H),6.95-6.8 4(m,0.5H),6.72-6.61(m,0.5H),6.51(s,1H),6.21-5.85(m,2H),5.78-5.52(m,1H),5.25-5.00 (m,1H),4.58-4.42(m,1H),4.06-3.88(m,3H),3.64-3.54(m,1H),3.42-3.28(s,1.5H),3.14-3. 06(m,0.5H),2.15-1.75(m,3H),1.32(d,J=7.6Hz,3H),0.80(d,J=7.2Hz,2H),0.64-0.48(m,2H).

[0260] Example 4 Preparation of Compound 004

[0261] (R)-1-(3-((5-cyclopropyl-2-((1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one

[0262]

[0263] Synthesis route and specific synthesis steps:

[0264]

[0265] Step 1: Synthesis of 4-nitro-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole (4i)

[0266] At room temperature, 4-nitro-1H-pyrazole 4g ​​(500mg, 4.4mmol), tetrahydro-2H-pyran-4-ol 4h (451mg, 4.4mmol), triphenylphosphine (1.4g, 5.3mmol) were dissolved in anhydrous tetrahydrofuran (20mL) and di-tert-butyl azodicarboxylate (1.32g, 5.7mmol) was added. The system was stirred at room temperature for 16 hours under nitrogen. Water (30mL) was added to quench the mixture and extracted with ethyl acetate (30mL*3). The organic layer was collected, dried, filtered, and dried. The residue was purified by thin layer column chromatography (ethyl acetate / petroleum ether: 10-50%) to obtain the title product 4i (600mg).

[0267] LCMS:Rt:1.26min;MS m / z(ESI):198.1[M+H] + .

[0268] Step 2: Synthesis of 1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-amine (4b)

[0269] 4-Nitro-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole 4i (600 mg, 3.2 mmol) was dissolved in methanol (10 mL) and wet palladium carbon (30 mg) was added at room temperature. The mixture was reacted at room temperature for 3 hours under a hydrogen atmosphere. It was then filtered and the filtrate was dried to give a crude product 4b (480 mg).

[0270] LCMS:Rt:0.376min;MS m / z(ESI):168.4[M+H] + .

[0271] Step 3: Synthesis of tert-butyl (R)-3-((5-cyclopropyl-2-(((1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate (4c)

[0272] At room temperature, (R)-tert-butyl 3-((2-chloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)aminopiperidine-1-carboxylate 4a (i.e., 1g in Example 1) (180 mg, 0.34 mmol), 1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-amine 4b (170 mg, 1.02 mmol), 1,1'-binaphthyl-2, 2'-Diphenylphosphine (21 mg, 0.034 mmol), cesium carbonate (326 mg, 1 mmol), tris(dibenzylideneacetone)dipalladium (31 mg, 0.034 mmol) and 1,4-dioxane (10 mL) were added to the reaction bottle, and the mixture was reacted under nitrogen at 120°C for 16 h. After the reaction was completed, the solvent was dried by spin drying, and the residue was purified by thin layer column chromatography (methanol / dichloromethane: 0-8%) to obtain the target product 4c (260 mg).

[0273] LCMS:Rt:1.306min; MS m / z(ESI):653.1[M+H] + .

[0274] Step 4: Synthesis of (R)-(5-cyclopropyl-4-(piperidin-3-ylamino)-2-((1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methanol (4d)

[0275] (3R)-tert-butyl 3-((5-cyclopropyl-2-((1-(tetrahydrofuran-3-yl)-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate 4c (200 mg, 0.31 mmol) was dissolved in dichloromethane (3 mL) at room temperature, 4M hydrochloric acid·dioxane (6 mL) was added and stirred for 1 hour, then dried by rotation, and the residue was dissolved in dichloromethane (3 mL) at room temperature, trifluoroacetic acid (3 mL) was added and stirred for 1 hour. The mixture was dried by rotation to give a crude product 4d (140 mg).

[0276] LCMS:Rt:0.875min; MS m / z(ESI):453.1[M+H] + .

[0277] Step 5: Synthesis of (R)-1-(3-((5-cyclopropyl-7-(hydroxymethyl)-2-((1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one (4f)

[0278] (R)-(5-cyclopropyl-4-(piperidin-3-ylamino)-2-((1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methanol 4d (140 mg, 0.31 mmol) and triethylamine (94 mg, 0.93 mmol) were dissolved in dichloromethane (10 mL) at room temperature, and the mixture was cooled to 0°C and propylene acid chloride 4e (35 mg, 0.6031 mmol) dissolved in dichloromethane (1 mL) was added and stirred for 2 hours. After the reaction was quenched with water, the solvent was dried by spin drying, and the residue was extracted with dichloromethane. The organic layer was collected, dried, and spin dried to give the crude product 4f (270 mg).

[0279] LCMS:Rt:1.127min;MS m / z(ESI):507.1[M+H] + .

[0280] Step 6: Synthesis of (R)-1-(3-((5-cyclopropyl-2-((1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one (004)

[0281] (R)-1-(3-((5-cyclopropyl-7-(hydroxymethyl)-2-((1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one 4f (157 mg, 0.31 mmol) was dissolved in tetrahydrofuran (2 mL) at room temperature and 2M sodium hydroxide (2 mL) was added. The mixture was stirred for 2 hours, the tetrahydrofuran was dried by spin drying, and then extracted with dichloromethane. The organic layer was dried by spin drying, dissolved in acetonitrile and purified by high performance liquid preparative chromatography to give the title compound 004 (58.0 mg).

[0282] Preparative high performance liquid chromatography: Waters (waters) 2767 preparative chromatograph, mobile phase A (Mobile phase A): 0.1% FA in water, mobile phase B (Mobile phase B): acetonitrile, chromatographic column: Sunfire (PrepC18 OBD 19*250mm 10μm), gradient (Gradient): 10% ACN run for 3min, 30%-35% ACN run to 14min, 95% ACN run to 17min, 10% ACN run to 21min to end purification and obtain the target compound.

[0283] LCMS:Rt:0.830min;MS m / z(ESI):477.2[M+H] + .

[0284] 1 H NMR(400MHz,DMSO-d6)δ10.60(br s,1H),8.44(s,1H),7.92(br s,1H),7.46(d,J=16.0Hz,1H),6.95-6.54(m,1H),6.45(s,1H),6.20-5.94(m, 1H),5.86-5.74(m,1H),5.74-5.45(m,1H),4.36-4.19(m,2H),3.96,3.93(two s,3H),3.85–3.51(m,5H),2.01–1.80(m,7H),1.75-1.47(m,2H),0.88-0.67(m,2H),0.58-0.36(m,2H).

[0285] Example 5 Preparation of Compound 005

[0286] 1-((3R)-3-((5-cyclopropyl-2-((1-(tetrahydrofuran-3-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one

[0287]

[0288] Synthesis route and specific synthesis steps:

[0289]

[0290] The preparation method was the same as that of Example 4, except that tetrahydrofuran-3-ol (5h) was used instead of tetrahydro-2H-pyran-4-ol (4h). The title compound 005 (57.98 mg) was purified by HPLC preparative chromatography.

[0291] Preparative high performance liquid chromatography: Waters 2767 preparative chromatograph, mobile phase A: 0.1% FA in water, mobile phase B: acetonitrile, chromatographic column: Sunfire (PrepC18 OBD 19*250mm 10μm), gradient: 10% ACN for 3 min, 30%-35% ACN for 14 min, 95% ACN for 17 min, and 10% ACN for 21 min to complete purification and obtain the target compound.

[0292] LCMS:Rt:1.09min;MS m / z(ESI):463.1[M+H] + .

[0293] 1H NMR (400MHz, DMSO-d6) δ10.62(d,J=7.2Hz,1H),8.48(s,1H),7.92(d,J=8.8H z,1H),7.47(d,J=15.2Hz,1H),6.92-6.58(m,1H),6.46(s,1H),6.13-5.99(m, 1H),5.84-5.49(m,2H),4.94-4.88(m,1H),4.32(s,1H),3.96–3.51(m,8H),2. 34–2.18(m,2H),1.96–1.59(m,5H),0.76(d,J=8.8Hz,2H),0.52–0.48(m,2H).

[0294] Example 6 Preparation of Compound 006

[0295] (R)-1-Acryloyl-3-((5-cyclopropyl-2-((1-oxetane-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine

[0296]

[0297] Synthesis route and specific synthesis steps:

[0298]

[0299] The preparation method was the same as that of Example 4, except that 3-oxetane alcohol (6b) replaced tetrahydro-2H-pyran-4-ol (4h) in Example 4, and purified by HPLC preparative chromatography to give the title compound 006 (24.48 mg).

[0300] Preparative high performance liquid chromatography: Waters 2767 preparative chromatograph, mobile phase A: 10 mmol% TFA in water, mobile phase B: acetonitrile, chromatographic column: Sunfire (Prep C18 OBD 19*250mm 10μm), gradient: 10% ACN for 4 min, 40%-45% ACN for 14 min, 95% ACN for 14 min, and 10% ACN for 17 min to complete purification and obtain the target compound.

[0301] LCMS:Rt:0.809min; MS m / z(ESI):449.1[M+H] + .

[0302] 1 H NMR(400MHz,DMSO)δ11.52-11.16(m,1H),9.89-9.51(m,1H),8.09-8.04(m,1 H),7.72-7.65(m,1H),6.97-6.89(m,1H),6.79-6.51(m,2H),6.20-5.70(m,2H ),5.54-5.51(m,1H),4.99-4.83(m,3H),4.39-4.01(m,3H),4.04-3.40(m,4H) ,2.10-1.88(m,2H),1.88-1.45(m,2H),1.00-0.79(m,2H),0.62-0.43(m,2H).

[0303] Example 7 Preparation of Compound 007

[0304] (R)-1-(3-((5-cyclopropyl-2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one

[0305]

[0306] Synthesis route and specific synthesis steps:

[0307]

[0308] The preparation method was the same as that of Example 4, except that 1-methylpiperidin-4-ol (7h) replaced tetrahydro-2H-pyran-4-ol (4h) in Example 4, and purified by HPLC preparative chromatography to give the title compound 007 (13.03 mg).

[0309] Preparative high performance liquid chromatography: Waters 2767 preparative chromatograph, mobile phase A: 0.1% FA in water, mobile phase B: acetonitrile, chromatographic column: Sunfire (PrepC18 OBD 19*250mm 10μm), gradient: 10% ACN for 3 min, 35%-40% ACN for 15 min, 95% ACN for 19 min, and 10% ACN for 22 min to complete purification and obtain the target compound.

[0310] LCMS:Rt:0.728min; MS m / z(ESI):490.2[M+H] + .

[0311] 1 H NMR (400MHz, DMSO-d6) δ10.61(s,1H),8.43(s,1H),7.91(s,1H),7.45(d,J=14.2Hz,1H),6.88(s,1H),6.45(s,1H),6.06(m,1H ),5.84–5.35(m,2H),4.43–3.34(m,8H),2.87(d,J=10.8Hz,2H),2.29–1.41(m,12H),0.76(d,J=7.8Hz,2H),0.58–0.37(m,2H).

[0312] Example 8 Preparation of Compound 008

[0313] 1-((3S,4R)-3-((5-cyclopropyl-2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-4-fluoropiperidin-1-yl)prop-2-en-1-one

[0314]

[0315] Synthesis route and specific synthesis steps:

[0316]

[0317] Step 1: Synthesis of 1-methyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine (8c)

[0318] Dissolve triphenylphosphine (34.8 g, 132.8 mmol) in anhydrous tetrahydrofuran (300 mL) and cool to 0°C. Then slowly inject DIAD (26.8 g, 132.8 mmol) and stir at 0°C for 10 minutes. A large amount of white precipitate is generated in the solution.

[0319] Compound 4-nitro-1H-pyrazole 8a (10.0 g, 88.5 mmol) and compound 1-methylpiperidin-4-ol 8b (11.2 g, 97.4 mmol) were dissolved in anhydrous tetrahydrofuran (100 mL) and then slowly added dropwise to the above solution. After the addition was complete, the ice bath was removed and the reaction solution was stirred at room temperature overnight.

[0320] Post-treatment: The reaction solution was concentrated under reduced pressure, and the obtained crude product was dissolved in ethyl acetate (150 mL), and then extracted with 3 mol / L hydrochloric acid solution (100 mL). The collected aqueous phase was adjusted to pH 9-10 with saturated potassium carbonate solution, and then extracted with ethyl acetate (200 mL x 2). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title product 8c (16.5 g).

[0321] LCMS:Rt:0.405min; MS m / z(ESI):211.1[M+H] + .

[0322] Step 2: Synthesis of 1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-amine (8d)

[0323] The compound 1-methyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine 8c (16.5 g, 78.6 mmol) was dissolved in ethanol (200 mL) at room temperature, and then 10% Pd / C (3.3 g) was added, and the reaction solution was stirred overnight under a hydrogen atmosphere. After the reaction was completed, the reaction solution was filtered and the filtrate was dried to obtain the title product 8d (12.8 g)

[0324] LCMS:Rt:1.47min;MS m / z(ESI):181.1[M+H] + .

[0325] Step 3: Synthesis of tert-butyl (3S, 4R)-3-((2-chloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-4-fluoropiperidine-1-carboxylate (8 g)

[0326] At room temperature, the compound 2,4-dichloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine 8e (357.0 mg, 1.0 mmol), the compound tert-butyl (3S,4R)-3-amino-4-fluoropiperidine-1-carboxylate 8f (545.0 mg, 2.5 mmol) and DIPEA (387.0 mg) were dissolved in ethanol (8 mL), and then the mixture was added to a jar and stirred at 120°C overnight. After the reaction was completed, the reaction solution was concentrated and dried. The crude product was separated and purified by normal phase column to obtain the title product 8g (256 mg). LCMS: Rt: 1.941 min; MS m / z (ESI): 540.2 [M+H] + .

[0327] Step 4: Synthesis of tert-butyl (3S,4R)-3-((5-cyclopropyl-2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-4-fluoropiperidine-1-carboxylate (8h)

[0328] The intermediate 1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-amine 8d (254 mg, 1.41 mmol), the intermediate tert-butyl (3S, 4R)-3-((2-chloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-4-fluoropiperidine-1-carboxylate 8g (256 mg, 0.47 mmol), Pd2(dba)3 (0.1 eq), BINAP (0.1 eq) and Cs2CO3 (3 eq) were added to dioxane (8 mL), and heated to 110° C. and stirred overnight under nitrogen protection. After the reaction was completed, the reaction solution was cooled to room temperature and filtered. The filtrate was directly purified by reverse phase column (Prep-HPLC (Boston Prime C18 150*30mm*5μm; A%: water (containing 0.225% FA); B%: ACN 10%-40%, 8-20min) to give the title product 8h (153.0 mg).

[0329] LCMS:Rt:1.279min;MS m / z(ESI):684.4[M+H] + .

[0330] Step 5: Synthesis of (5-cyclopropyl-4-(((3S,4R)-4-fluoropiperidin-3-yl)amino)-2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methanol (8i)

[0331] The intermediate tert-butyl (3S, 4R)-3-((5-cyclopropyl-2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-4-fluoropiperidine-1-carboxylate 8h (153 mg, 0.22 mmol) was dissolved in TFA (3 mL) and then stirred at room temperature for 1 h. After the reaction was completed, the mixture was dried by rotation, and the title product 8i (185 mg, purity: 66.8%) was directly used for the next step reaction.

[0332] LCMS:Rt:1.171min;MS m / z(ESI):484.1[M+H]+ .

[0333] Step 6: 5-Cyclopropyl-N 4 -((3S,4R)-4-Fluoropiperidin-3-yl)-N 2 Synthesis of -(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine-2,4-diamine (8j)

[0334] (5-cyclopropyl-4-(((3S,4R)-4-fluoropiperidin-3-yl)amino)-2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methanol 8i (185 mg) was dissolved in a mixture of acetone (3 mL) and H2O (3 mL), solid potassium carbonate was added to adjust the pH to 9-10, and then the reaction solution was heated to 40°C and stirred overnight. LCMS detected that the intermediate 8i was completely converted to the title product 8j, and the reaction solution was directly subjected to the next step reaction.

[0335] LCMS:Rt:1.45min;MS m / z(ESI):454.1[M+H] + .

[0336] Step 7: Synthesis of 1-((3S,4R)-3-((5-cyclopropyl-2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-4-fluoropiperidin-1-yl)prop-2-en-1-one (008)

[0337] The reaction solution of the previous step was cooled to 0°C, and potassium carbonate solid was added to adjust the pH to 9-10, and then 8k acrylate chloride (91 mg, 0.66 mmol) was dissolved in acetone (1 mL) and added dropwise to the reaction solution. After the addition was completed, the reaction solution was stirred at 0°C for 1 h, and LCMS detected that the raw material disappeared. After the reaction was completed, the reaction solution was filtered and directly sent to the preparation (Prep-HPLC (Boston Prime C18 150*30mm*5μm; A%: water (containing 0.225% FA); B%: ACN 13%-43%, 8-15min) for purification to obtain the target product 008 (9.58 mg).

[0338] LCMS:Rt:5.287min; MS m / z(ESI):508.7[M+H] + .

[0339] 1H NMR (400MHz, DMSO-d6) δ10.66(s,1H),8.50(s,1H),7.91(s,1H),7.43(d,J=6.0Hz,1H),6.95-6.76(m,1H ),6.47(s,1H),6.16-6.10(m,1H),5.88-5.61(m,2H),5.13-4.94(m,1H),4.89(s,1H),3.99-3.46(m,5H), 2.84(d,J=10.8Hz,2H),2.19(s,3H),2.04-1.72(m,9H),0.87-0.69(m,2H),0.60-0.51(m,1H),0.46-0.40(m,1H).

[0340] Example 9 Preparation of Compound 009

[0341] 1-((2S,5R)-5-((5-cyclopropyl-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one

[0342]

[0343] Synthesis route and specific synthesis steps:

[0344]

[0345] Step 1: Synthesis of benzyl (2S,5R)-5-((2-chloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidine-1-carboxylate (9c)

[0346] 2,4-Dichloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine (9a, 500 mg, 1.39 mmol), (2S,5R)-5-amino-2-methylpiperidine-1-carboxylic acid benzyl ester (9b, 346 mg, 1.39 mmol) and DIPEA (448 mg, 3.48 mmol) were added to an isopropanol (10.0 mL) solution at room temperature, and the reaction system was heated to 110°C and stirred at this temperature for 16 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by column chromatography (ethyl acetate / petroleum ether = 1 / 10) to obtain 9c (385 mg, yield: 48.6%).

[0347] LCMS: Rt: 2.157min; MS m / z (ESI): 570.2[M+H];

[0348] 1 H NMR(400M Hz,DMSO-d6)δ7.45-7.39(m,4H),7.35-7.32(m,1H),6.77(s,1H),5.72(d,J=8.0Hz,1H),5.46(s ,2H),5.20(dd,J=17.6,12.4Hz,2H),4.60-4.55(m,2H),4.26-4.20(m,1H),3.54(t,J=8.0Hz,2H) ,2.73(t,J=11.6Hz,1H),2.21(s,1H),2.12-2.08(m,1H),1.93-1.88(m,1H),1.70-1.65(m,2H), 1.31-1.28(m,1H),1.25(d,J=6.8Hz,3H),0.93(t,J=8.0Hz,3H),0.72-0.71(m,2H),0.03(s,9H).

[0349] Step 2: Synthesis of benzyl (2S,5R)-5-((5-cyclopropyl-2-((1-methyl-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidine-1-carboxylate (9e)

[0350] Under nitrogen protection at room temperature, Pd2(dba)3 (32.0 mg, 0.035 mmol) and BINAP (21.8 mg, 0.035 mmol) were added to a solution of 9c (200 mg, 0.351 mmol), 1-methyl-1H-pyrazole-4-amino (9d, 40.8 mg, 0.421 mmol) and Cs2CO3 (285.9 mg, 0.878 mmol) in 1,4-dioxane (10.0 mL), and the reaction system was heated to 100°C and stirred for 16 hours, then cooled to room temperature. The mixture was filtered and the filter cake was washed with ethyl acetate (30.0 mL). The solution was concentrated under reduced pressure and purified by Chem-flash (acetonitrile / 0.1% trifluoroacetic acid) to obtain 9e (195 mg, yield: 88.0%).

[0351] LCMS: Rt: 2.148min; MS m / z (ESI): 631.3[M+H].

[0352] Step 3: 5-Cyclopropyl-N 2 -(1-methyl-1H-pyrazol-4-yl)-N 4Synthesis of -((3R,6S)-6-methylpiperidin-3-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine-2,4-diamine (9f)

[0353] Under nitrogen protection at room temperature, Pd / C (5 mg) was added to 9e (195 mg, 0.309 mmol) in methanol (5.0 mL), and the reaction system was stirred at room temperature for 16 hours under hydrogen. The reaction mixture was filtered under reduced pressure, the filter cake was washed with methanol (20.0 mL), the filtrate was combined, evaporated to dryness under reduced pressure, and the residue was purified by Chem-flash (acetonitrile / 0.1% trifluoroacetic acid) to obtain 9f (136 mg, yield: 88.3%).

[0354] LCMS: Rt: 1.212min; MS m / z (ESI): 497.3 [M+H].

[0355] Step 4: 5-cyclopropyl-N 2 -(1-methyl-1H-pyrazol-4-yl)-N 4 Synthesis of -((3R,6S)-6-methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidine-2,4-diamine (9 g)

[0356] At 0°C, trifluoroacetic acid (1 mL) was added to a dichloromethane solution (5.0 mL) of 9f (136 mg, 0.273 mmol), and the reaction solution was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting yellow oil was added to a methanol solution (5.0 mL) of DIPEA (1.0 mL), and the reaction solution was stirred at 50°C for 6 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by Chem-flash (acetonitrile / 0.1% trifluoroacetic acid) to obtain 9g (95 mg, yield: 94.8%).

[0357] LCMS: Rt: 1.124min; MS m / z (ESI): 367.2[M+H].

[0358] Step 5: Synthesis of 1-((2S,5R)-5-((5-cyclopropyl-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (009)

[0359] 9g (95mg, 0.259mmol) was dissolved in a mixture of tetrahydrofuran (2.0mL) and H2O (3.0mL), and potassium phosphate solid (137.5mg, 0.648mmol) was added. Acryloyl chloride (9h, 28.2mg, 0.311mmol) was added dropwise to the reaction solution under stirring at 0℃, and stirred at room temperature for 2 hours. The reaction solution was filtered, and the filtrate was prepared and purified by Prep-HPLC (formic acid system) (A%: water (containing 0.225% FA); B%: ACN 13%-43%, 8-10min) to obtain 009 (18.35mg, yield: 16.8%).

[0360] LCMS: Rt: 4.416min; MS m / z (ESI): 421.5[M+H];

[0361] 1 H NMR(400M Hz,DMSO-d6)δ10.59(s,1H),8.40(s,1H),7.81(s,1H),7.38(s,1H),6.92(brs,1H),6.4 3(s,1H),6.11-6.07(m,1H),5.87(d,J=8.4Hz,1H),5.64(brs,1H),4.79-4.28(m,2H),4. 13(brs,2H),3.71(s,3H),2.76-2.70(m,1H),2.03-1.99(m,1H),1.87-1.85(m,2H),1.72 -1.67(m,2H),1.23(s,3H),0.83(d,J=8.0Hz,1H),0.61-0.57(m,1H),0.47-0.43(m,1H).

[0362] Example 10 Preparation of Compound 010

[0363] (R)-1-(3-((5-cyclopropyl-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one

[0364]

[0365] Synthesis route and specific synthesis steps:

[0366]

[0367] Step 1: Synthesis of (R)-tert-butyl 3-((2-chloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidine-1-carboxylate (10c)

[0368] 10a (500 mg, 1.4 mmol), (R)-3-aminopyrrolidine-1-carboxylic acid tert-butyl ester (10b, 313 mg, 1.68 mmol) and DIPEA (543 mL, 4.2 mmol) were added to an isopropanol (10.0 mL) solution at room temperature. The reaction system was sealed and heated to 110°C and stirred at this temperature for 16 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by column (petroleum ether / ethyl acetate = 4 / 1) to obtain 10c (450 mg, yield: 49.3%). LCMS: Rt: 1.320 min; MS m / z (ESI): 508.3 [M+H].

[0369] Step 2: Synthesis of tert-butyl ((R)-3-((5-cyclopropyl-2-((1-methyl-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidine-1-carboxylate (10e)

[0370] At room temperature, under nitrogen protection, Pd2(dba)3 (72.3 mg, 0.079 mmol) and BINAP (49.1 mg, 0.079 mmol) were added to a solution of 10c (400 mg, 0.79 mmol), 1-methyl-1H-pyrazol-4-amine (10d, 92 mg, 0.94 mmol) and cesium carbonate (772 mg, 2.37 mmol) in 1,4-dioxane (30 mL). The reaction system was heated to 100°C and stirred for 16 hours, then cooled to room temperature. Water (30 ml) was added to quench the reaction, and the mixture was extracted with ethyl acetate (50.0 ml*2). The organic phase was concentrated and purified by column (petroleum ether / ethyl acetate = 2 / 1) to obtain 10e (361 mg, yield: 80.4%).

[0371] LCMS: Rt: 1.474min; MS m / z (ESI): 569.3[M+H];

[0372] 1H NMR (400MHz, DMSO-d6) δ8.82 (s, 1H), 8.00 (s, 1H), 7.60 (s, 2H), 6.74 (d, J = 0. 8Hz,1H),6.07(s,1H),5.45(s,2H),4.82-4.75(m,1H),3.90(s,3H),3.79-3. 76(m,1H),3.59(t,J=8.0Hz,3H),3.48(s,1H),3.43(s,1H),2.34(s,1H),2.1 4(s,1H),1.51(d,J=8.8Hz,9H),0.94-0.91(m,4H),0.62(s,2H),0.00(s,9H).

[0373] Step 3: (R)-5-cyclopropyl-N 2 -(1-methyl-1H-pyrazol-4-yl)-N 4 Synthesis of -(pyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidine-2,4-diamine (10f)

[0374] 10e (361 mg, 0.63 mmol) was added to a dichloromethane solution (3.0 mL) of trifluoroacetic acid (1.0 mL) at room temperature, and the reaction solution was stirred at room temperature for 2 hours to obtain a single product. The reaction solution was concentrated under reduced pressure and added to a lithium hydroxide solution (100 mg, 2.3 mmol) of tetrahydrofuran / water (4.0 mL / 2 mL), and the reaction solution was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure to obtain 10f (190 mg, yield: 88.8%).

[0375] LCMS: Rt: 1.245min; MS m / z (ESI): 339.5 [M+H].

[0376] Step 4: Synthesis of (R)-1-(3-((5-cyclopropyl-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one (010)

[0377] 10f (190 mg, 0.56 mmol) was dissolved in a mixture of tetrahydrofuran (4.0 mL) and H2O (2.0 mL), potassium phosphate solid (297 mg, 1.4 mmol) was added, and the mixture was stirred at room temperature. Acryloyl chloride 10 g (71 mg, 0.78 mmol) was added dropwise, and the mixture was stirred at 0°C for 2 hours. The mixture was filtered and purified (Prep-HPLC (Boston Prime C18 150*30mm*5μm; A%: water (containing 0.225% FA); B%: ACN 13%-43%, 8-10 min) to obtain 010 (25.59 mg, yield: 11.2%).

[0378] LCMS: Rt: 6.005min; MS m / z (ESI): 393.2[M+H].

[0379] 1 H NMR(400M Hz, DMSO-d6) δ10.83(brs,1H),8.88(brs,1H),7.84(d,J=2.4Hz,1H),7.49(d,J=3.6H z,1H),6.67-6.54(m,1H),6.51(s,1H),6.18-6.12(m,1H),5.68(t,J=12.8Hz,1H),4. 81-4.69(m,1H),4.03-3.99(m,1H),3.79(s,3H),3.74-3.59(m,2H),3.54-3.44(m,2H ),2.34-2.17(m,1H),2.07-1.99(m,2H),0.79(t,J=2.8Hz,2H),0.50(d,J=2.4Hz,2H).

[0380] Example 11 Preparation of Compound 011

[0381] (E)-2-((2S,5R)-5-((5-cyclopropyl-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidine-1-carbonyl)-4,4-dimethylpent-2-enenitrile

[0382]

[0383] Synthesis route and specific synthesis steps:

[0384]

[0385] Step 1: Synthesis of 2-cyano-4,4-dimethyl-2-pentenoic acid (11a)

[0386] Dissolve cyanoacetic acid (5.00 g, 58.0 mmol) in anhydrous methanol (50.0 mL), add trimethylacetaldehyde (5.00 g, 58.0 mmol) and sodium hydroxide (4.64 g, 116 mmol), and stir at 40°C for 1 hour. The reaction solution is concentrated under reduced pressure, and the residue is purified by column chromatography (methanol / dichloromethane = 1 / 10) to obtain intermediate 11a (4.2 g, 65.0%).

[0387] 1 H NMR (400MHz, CDCl3) δ = 7.74 (d, J = 0.8Hz, 1H), 1.40-1.30 (s, 9H).

[0388] Step 2: Synthesis of (E)-2-((2S,5R)-5-((5-cyclopropyl-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidine-1-carbonyl)-4,4-dimethylpent-2-enenitrile (011)

[0389] The intermediate 009 (250 mg, 682 μmol) and 2-cyano-4,4-dimethyl-2-pentenoic acid 11a (115 mg, 750 μmol) were dissolved in N,N-dimethylformamide (4.0 mL), triethylamine (207 mg, 2.1 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (259 mg, 682 μmol) were added, and stirred at 25°C for 16 hours. The reaction solution was filtered and separated and purified by Prep-HPLC (Boston Prime C18150*30mm*5μm; mobile phase: A%: water (containing 0.225% FA); B%: ACN22%-52%, 9 min) to obtain 011 (5.9 mg, yield: 1.7%).

[0390] LCMS: Rt: 3.573min; MS m / z(ESI): 502.5[M+H].

[0391] 1 H NMR (400MHz, METHANOL-d4) δ = 7.89 (s, 1H), 7.49 (s, 1H), 6.79 (br s, 1H), 6.47 (d, J = 1.2Hz, 1H), 4.22 (br s,1H),3.85(s,3H),3.23-2.87(m,1H),2.11-1.78(m,5H),1.45-1.10(m,12H),0.96-0.82(m,2H),0.71-0.49(m,1H),0.59(br d,J=5.3Hz,1H)

[0392] Example 12 Preparation of Compound 012

[0393] 1-((2S,5R)-5-((5-cyclopropyl-2-((1-(trifluoromethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one

[0394]

[0395] Synthesis route and specific synthesis steps:

[0396]

[0397] Step 1: Synthesis of (2S,5R)-benzyl 5-((2-chloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidine-1-carboxylate (12c)

[0398] 2,4-Dichloro-5-cyclopropyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine 12a (2.0 g, 5.6 mmol) was dissolved in anhydrous isopropanol (30.0 mL), diisopropylethylamine (1.4 g, 11.2 mmol) and (2S,5R)-5-amino-2-methylpiperidine-1-carboxylic acid benzyl ester 12b (1.52 g, 6.1 mmol) were added, and stirred at 80°C for 16 hours. After cooling, the reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate / petroleum ether = 1 / 4) to obtain intermediate 12c (1.3 g, 40.9%).

[0399] LCMS: Rt: 1.116min; MS m / z(ESI): 570.2[M+H].

[0400] Step 2: Synthesis of (2S,5R)-benzyl-5-((5-cyclopropyl-2-((1-(trifluoromethyl)-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidine-1-carboxylate (12e)

[0401] Intermediate 12c (700 mg, 1.2 mmol) and 1-(trifluoromethyl)-1H-pyrazol-4-amine 12d (253 mg, 1.4 mmol) were dissolved in anhydrous dioxane (30.0 mL), tri(dibenzylideneacetone)dipalladium (112 mg, 123 μmol), dinaphthyl diphenylphosphine (76 mg, 123 μmol) and cesium carbonate (1.0 g, 3.1 mmol) were added, and stirred at 100 ° C for 16 hours under nitrogen atmosphere. After cooling, water (10.0 mL) was added for dilution, and ethyl acetate (20.0 mL*2) was used for extraction. The organic phase was washed with saturated sodium chloride aqueous solution (20.0 mL), dried, filtered and concentrated. The residue was purified by column chromatography (ethyl acetate / petroleum ether = 1 / 2) to obtain intermediate 12e (720 mg, 85.6%).

[0402] LCMS: Rt: 1.240min; MS m / z(ESI): 685.3[M+H].

[0403] Step 3: 5-Cyclopropyl-N 4 -((3R,6S)-6-methylpiperidin-3-yl)-N 2 Synthesis of -(1-(trifluoromethyl)-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine-2,4-diamine (12f)

[0404] Intermediate 12e (370 mg, 540 μmol) was dissolved in anhydrous methanol (15.0 mL), and wet palladium carbon (40 mg, 10% Pd) was added. The mixture was stirred at 25°C for 16 hours under a hydrogen atmosphere (15 psi). The reaction solution was filtered and concentrated to obtain a crude product 12f (270 mg), which was used directly in the next step without purification.

[0405] LCMS: Rt: 1.239min; MS m / z(ESI): 551.3[M+H].

[0406] Step 4: 5-Cyclopropyl-N 4 -((3R,6S)-6-methylpiperidin-3-yl)-N 2 Synthesis of -(1-(trifluoromethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine-2,4-diamine (12 g)

[0407] The crude product 12f (270 mg, 490 μmol) was dissolved in anhydrous dichloromethane (10.0 mL). Trifluoroacetic acid (2.0 mL) was added, and the mixture was stirred at 25 °C for 6 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was dissolved in methanol (6.0 mL) and water (3.0 mL). Lithium hydroxide (21 mg, 490 μmol) was added, and the mixture was heated to 50 °C and stirred for 2 hours. The reaction solution was cooled, filtered, and concentrated. After adding water to the residue, a solid precipitated. The green solid crude product 12 g (260 mg) was obtained by filtration and used directly in the next step without purification.

[0408] LCMS: Rt: 2.543min; MS m / z(ESI): 421.1[M+H].

[0409] Step 5: Synthesis of 1-((2S,5R)-5-((5-cyclopropyl-2-((1-(trifluoromethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (012)

[0410] At 0°C, 12 g (260 mg, 495 μmol) of crude product was dissolved in tetrahydrofuran (4.0 mL) and water (2.0 mL). Potassium phosphate (228 mg, 990 μmol) and acryloyl chloride (45 mg, 495 μmol) were added, and the temperature was raised to 25°C and stirred for 2 hours. The mixture was diluted with water (10.0 mL), extracted with ethyl acetate (15.0 mL*2), and the organic phase was washed with saturated sodium chloride aqueous solution (15.0 mL), dried, filtered, and concentrated. The residue was purified by preparative purification (Prep-HPLC (Boston Prime C18 150*30mm*5μm; A%: water (containing 0.225% FA); B%: ACN 22%-52%)) to obtain product 012 (25.4 mg, 10.8%).

[0411] LC-MS: Rt: 2.892min; MS m / z (ESI): 475.2[M+H];

[0412] 1H NMR (400MHz, DMSO-d6)δ=10.77(br s,1H),8.95(s,1H),8.39(br s,1H),7.89(br s,1H),6.78(br s,1H),6.52(s,1H),6.18-5.94(m,2H),5.63(br s,1H),4.92-3.93(m,3H),3.17-2.72(m,1H),2.04(br s,1H),1.88(br s,2H),1.70(br s,2H),1.22(br s,3H),0.85(br d,J=7.8Hz,2H),0.66-0.37(m,1H),0.66-0.37(m,2H).

[0413] Example 13 Preparation of Compound 013

[0414] (R)-1-(4-((5-chloro-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)nonyl-1-yl)prop-2-en-1-one

[0415]

[0416] Synthesis route and specific synthesis steps:

[0417]

[0418] Step 1: Synthesis of tert-butyl 4-((2,5-dichloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)azepane-1-carboxylate (13c)

[0419] At room temperature, NaH (60 mg, 1.5 mmol) was added in batches to a solution of tert-butyl 4-hydroxyazepane-1-carboxylate 13b (323 mg, 1.5 mmol) in anhydrous DMSO (10.0 mL). The mixture was stirred for 15 minutes after addition, and then 2,4,5-trichloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine 13a (527 mg, 1.5 mmol) was added. The reaction system was heated to 55°C and reacted for 15 minutes. The mixture was cooled to room temperature, and a saturated ammonium chloride solution (50.0 mL) was added to the reaction solution. The mixture was extracted with dichloromethane three times (50.0 mL*3). The organic phases were combined, washed once with water (50.0 mL) and saturated brine (5.0 mL), and the organic phase was dried and filtered over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure and purified by a silica gel column (ethyl acetate: petroleum ether = 1:10) to obtain 13c (620.4 mg, yield: 78%).

[0420] LCMS: Rt: 1.775min; MS m / z(ESI): 533.5[M+H].

[0421] Step 2: Synthesis of tert-butyl 4-((5-chloro-2-((1-methyl-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)azepane-1-carboxylate (13e)

[0422] Under nitrogen protection at room temperature, 13c (620.4 mg, 1.17 mmol), 1-methyl-1H-pyrazol-4-amine 13d (170 mg, 1.76 mmol) and Cs2CO3 (763 mg, 2.34 mmol) were added to a solution of 1,4-dioxane (20.0 mL). The reaction system was heated to 100 °C and stirred for 16 hours, then cooled to room temperature. The filter cake was washed with tetrahydrofuran (50.0 mL), concentrated under reduced pressure, and purified to obtain the product 13e (595.8 mg, yield: 86.2%).

[0423] LCMS: Rt: 1.480min; MS m / z(ESI): 592.2[M+H].

[0424] 1 H NMR (400MHz, CDCl3) δ7.84(s,1H),7.60(s,1H),6.87(s,1H),6.64(d,J=7.6Hz,1H),5.59-5.54(m,1H),5.49(s,2H),3.97(s,3H),3.83-3.6 6(m,2H),3.60(t,J=8.0Hz,2H),3.49-3.39(m,2H),2.23-2.08(m,4H),1.93-1.82(m,2H),1.54(s,9H),0.98(t,J=8.4Hz,2H),0.00(s,9H).

[0425] Step 3: Synthesis of ((4-(azepan-4-oxy)-5-chloro-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methanol (13f)

[0426] At room temperature, TFA (12.0 mL) was added to a solution of intermediate 13e (6.0 mL). After stirring at room temperature for 2 hours, the reaction solution was concentrated to dryness under reduced pressure, and the obtained crude product 13f was directly used in the next reaction.

[0427] LCMS: Rt: 0.763min; MS m / z(ESI): 392.0[M+H].

[0428] Step 4: Synthesis of 1-(4-((5-chloro-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)azepan-1-yl)prop-2-en-1-one (013 and 013-1)

[0429] The crude product 13f (1 mmol) was dissolved in a mixed solvent of acetone (12.0 ml) and water (8.0 mL), and the pH value was adjusted to 10 with potassium carbonate, heated to 35°C, and stirred for 16 hours. The reaction solution was cooled to 0°C, and then acryloyl chloride (126 mg, 1.4 mmol) was dissolved in acetone (1.0 mL) and added dropwise to the reaction solution. After the addition was completed, the reaction solution was stirred at 0°C for 1 hour. The reaction solution was filtered and dried for purification (Prep-HPLC (Boston Prime C18 150*30mm*5μm; A%: water (containing 0.225% FA); B%: ACN 22%-52%)) to obtain the product.

[0430] 013 (14.28 mg, yield: 6.9%):

[0431] LCMS: Rt: 6.567min; MS m / z (ESI): 416.1[M+H];

[0432] 1 H NMR(400MHz,DMSO-d6)δ11.47(s,1H),9.01(s,1H),7.83(s,1H),7.49(s,1H),7. 03(d,J=1.6Hz,1H),6.84-6.74(m,1H),6.18-6.11(m,1H),5.66(dd,J=10.4,2.4H z,1H),5.47(d,J=4.8Hz,1H),3.81(s,3H),3.79-3.76(m,2H),3.63-3.57(m,1H) ,3.47-3.40(m,1H),2.04(t,J=4.8Hz,3H),1.98-1.91(m,1H),1.84-1.76(m,2H).

[0433] 013-1 (13.82 mg, yield: 6.7%):

[0434] LCMS: Rt: 6.567min; MS m / z (ESI): 416.1[M+H];

[0435] 1H NMR(400MHz,DMSO-d6)δ11.47(s,1H),9.01(s,1H),7.83(s,1H),7.49(s,1H),7 .03(d,J=2.0Hz,1H),6.84-6.74(m,1H),6.18-6.11(m,1H),5.66(dd,J=8.0,2.4 Hz,1H),5.50-5.46(m,1H),3.80(s,3H),3.76-3.71(m,2H),3.64-3.57(m,1H), 3.46-3.39(m,1H),2.04(t,J=5.2Hz,3H),1.98-1.91(m,1H),1.84-1.78(m,2H).

[0436] Example 14 Preparation of Compound 014

[0437] 1-((3S,4R)4-Fluoro-3-((2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)-piperidin-1-yl)prop-2-en-1-one

[0438] 1-((3S,4R)-4-fluoro-3-((2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)piperidin-1-yl)prop-2-en-1-one

[0439]

[0440] Synthesis route and specific synthesis steps:

[0441]

[0442] Step 1: Synthesis of tert-butyl 1-((3S,4R)4-fluoro-3-((2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7((2-(trimethylsilyl)ethoxy)methyl)-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)-piperidine-1-carboxylate (14c)

[0443] Under nitrogen protection at room temperature, tert-butyl 1-((3S,4R)4-fluoro-3-((2-chloro)-7((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)-tetrahydropyrrole-1-carboxylate 14a (125 mg, 0.25 mmol) and 1-(1-methylpiperidin-4-yl)-1H-pyrazole-4-amine 14b (67.5 mg, 0.38 mmol) were dissolved in 1,4-dioxane (10.0 mL). Then, Pd2(dba)3 (22.8 mg, 0.025 mmol), BINAP (31.1 mg, 0.2 mmol) and cesium carbonate (162.8 mg, 0.5 mmol) were added under stirring, and the reaction system was heated to 120°C. After stirring for 12 hours, it was cooled to room temperature and filtered. The filter cake was washed with ethyl acetate (60.0 mL), and the filtrate was concentrated under reduced pressure and purified (dichloromethane: methanol = 15:1) to obtain 14c (120 mg, yield: 74.5%).

[0444] LCMS: Rt: 1.307min; MS m / z (ESI): 645.3[M+H].

[0445] 1 H NMR(400MHz,DMSO-d6)δ9.24(s,1H),8.12(brs,1H),7.68(s,1H),7.21(d,J=3.6Hz,1H ),6.42(d,J=3.4Hz,1H),5.60(s,3H),5.18(d,J=46.3Hz,1H),4.48(s,1H),4.24(s,1H ),4.04(s,1H),3.63(t,J=8.0Hz,2H),3.04(d,J=28.0Hz,3H),2.41(s,3H),2.20(s,3H ),2.11(s,6H),1.47(s,2H),1.35(s,1H),1.08(s,6H),0.99–0.92(m,2H),0.00(s,9H).

[0446] Step 2: Synthesis of 1-((3S,4R)4-fluoro-3-((2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)-piperidine (14d)

[0447] Trifluoroacetic acid (5.0 mL) was added to a dichloromethane solution (5.0 mL) of 14c (125 mg, 0.19 mmol) at 0°C and stirred at room temperature for 16 hours. After concentration, the crude product was dissolved in a mixture of THF / H2O (6.0 mL, 2:1), and LiOH (125.0 mg, 5.2 mmol) was added and stirred at room temperature for 1 hour to obtain the crude product which was filtered and purified to give 14d (87 mg, yield: 99%).

[0448] LCMS: Rt: 0.427min; MS m / z(ESI): 415.2[M+H].

[0449] Step 3: Synthesis of 1-((3S,4R)4-fluoro-3-((2-((1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)-piperidin-1-yl)prop-2-en-1-one (014)

[0450] At 0°C, 14d (87 mg, 0.21 mmol) was added to a mixture of tetrahydrofuran (2.0 mL) and H2O (1.0 mL), and potassium phosphate (233.5 mg, 1.1 mmol) was added. After stirring for 10 minutes, a solution of acryloyl chloride (29.0 mg, 0.32 mmol) dissolved in tetrahydrofuran (1.0 mL) was slowly added dropwise. After stirring for 2 hours, the mixture was filtered and the filtrate was prepared and purified (Prep-HPLC (Boston Prime C18 150*30mm*5μm; A%: water (containing 0.225% FA); B%: ACN 20%-50%)) to obtain the product 014 (35.11 mg, yield: 35.7%).

[0451] LCMS: Rt: 5.937min; MS m / z (ESI): 469.3[M+H].

[0452] 1 H NMR(400MHz,DMSO-d6)δ11.32(s,1H),8.92(s,1H),8.23(s,1H),7.95(s,1H ),7.53(s,1H),7.02-6.77(m,1H),6.66-6.41(m,1H),6.27-5.90(m,1H),5. 78-5.37(m,1H),5.19(d,J=48.3Hz,1H),3.96(m,8H),3.72(d,J=32.5Hz,1H ),3.41(s,1H),2.97(d,J=8.2Hz,1H),2.29(d,J=24.4Hz,4H),2.00(s,4H).

[0453] Example 15 Preparation of Compound 015

[0454] 1-((3S,4R)-4-fluoro-3-((2-((1-((R)-tetrahydrofuran-3-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)piperidin-1-yl)prop-2-en-1-one

[0455]

[0456] Synthesis route and specific synthesis steps:

[0457]

[0458] Step 1: Synthesis of tert-butyl (3S,4R)4-fluoro-3-((2-(((1-((R)-tetrahydrofuran-3-yl)-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)piperidine-1-carboxylate (15 g)

[0459] Intermediate 15e (100 mg, 0.20 mmol), (R)-1-(tetrahydrofuran-3-yl)-1H-pyrazol-4-amine 15f (61 mg, 0.40 mmol), Pd(dppf)Cl2 (15 mg, 0.02 mmol), XantPhos (12 mg, 0.02 mmol) and cesium carbonate (130 mg, 0.40 mmol) were added to 1,4-dioxane (2.0 mL), and the reaction solution was stirred at 95 ° C for 16 hours under argon protection. After cooling, the reaction solution was filtered with diatomaceous earth, and the filter cake was washed with ethyl acetate (10.0 mL). The filtrate was concentrated and purified under reduced pressure (petroleum ether / ethyl acetate = 2:1) to obtain 15g (150 mg, yield: 90%).

[0460] LCMS: Rt: 0.73min; MS m / z(ESI): 618.4[M+H].

[0461] 1H NMR (400MHz, CDCl3) δ8.04(s,1H),7.65(s,1H),6.96(d,J=3.6Hz,1H),6.67(brs,1H),6.52(d,J= 3.6Hz,1H),5.57(s,2H),5.46-5.37(m,1H),5.19-5.03(m,2H),4.25-4.20(m,1H),4.17-4.11(m, 1H),4.05-4.01(m,3H),3.90-3.69(m,2H),3.63-3.59(m,3H),2.56-2.50(m,1H),2.43-2.38(m,1 H),2.31-2.21(m,1H),2.03-1.89(m,1H),1.59-1.35(m,9H),0.98(t,J=8.8Hz,2H),0.00(s,9H).

[0462] Step 2: Synthesis of 4-(((3S,4R)-4-fluoropiperidin-3-yl)oxy)-N-(1-((R)-tetrahydrofuran-3-yl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine (15h)

[0463] 15g (150mg, 0.24mmol) was added to trifluoroacetic acid (3.0mL) at 25°C, and the reaction solution was stirred at 25°C for 30 minutes. The reaction solution was concentrated under reduced pressure and added to a solution of lithium hydroxide monohydrate in tetrahydrofuran (3.0mL) / water (0.5mL) (pH=12), and the reaction solution was stirred at 25°C for 30 minutes. The mixture was concentrated under reduced pressure, diluted with 5.0mL of water, and extracted with ethyl acetate (5.0mL*3). The organic phase was concentrated to obtain 15h (120mg, yield:>99%).

[0464] LCMS: Rt: 1.15min; MS m / z(ESI): 388.2[M+H].

[0465] Step 3: Synthesis of 1-((3S,4R)-4-fluoro-3-((2-((1-((R)-tetrahydrofuran-3-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)piperidin-1-yl)prop-2-en-1-one (015)

[0466] 15h (100 mg, 0.26 mmol) was dissolved in a mixture of tetrahydrofuran (0.5 mL) and water (0.3 mL), potassium phosphate solid (110 mg, 0.52 mmol) was added, and the mixture was stirred at 25°C. A solution of acryloyl chloride (23 mg, 0.26 mmol) in tetrahydrofuran (0.5 mL) was added dropwise, and stirred for 1 hour. The reaction solution was concentrated under reduced pressure and purified (Prep-HPLC (Boston Prime C18 150*30mm*5μm; A%: water (containing 0.225% FA); B%: ACN 15%-40%)) to obtain 015 (8.5 mg, yield: 7%).

[0467] LCMS: Rt: 5.07min; MS m / z(ESI): 442.3[M+H].

[0468] 1 H NMR(400M Hz,DMSO-d6)δ11.31(s,1H),8.94(s,1H),7.94(s,1H),7.54-7.50(m,1H),6.94(s,1H),6.90-6.49(m,1H),6.21-6.16(m,1H),6.11-5.93(m ,1H),5.73-5.41(m,2H),5.24-4.94(m,2H),4.06-3.96(m,3H),3.88- 3.60(m,5H),2.39-2.32(m,1H),2.26-2.06(m,1H),2.08-1.76(m,2H).

[0469] Example 16 Preparation of Compound 016

[0470] 1-((3S,4R)-4-fluoro-3-((2-((1-((R)-1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)piperidin-1-yl)prop-2-en-1-one

[0471]

[0472] Synthesis route and specific synthesis steps:

[0473]

[0474] Step 1: Synthesis of tert-butyl (3S,4R)-3-((2-chloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)-4-fluoropyridine-1-carboxylate (16c)

[0475] At room temperature, NaH (91 mg, 2.27 mmol) was added in batches to a solution of (3S, 4R)-4-fluoro-3-hydroxypiperidine-1-carboxylic acid tert-butyl ester 16b (500 mg, 2.27 mmol) in DMSO (15.0 mL). The mixture was stirred for 15 minutes. Then, 2,4-dichloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine 16a (720 mg, 2.2 7mmol) was added, the reaction system was heated to 55°C, reacted for 15 minutes, 50.0mL of saturated NH4Cl solution was added to the reaction solution, extracted with dichloromethane three times (50.0mL*3), the organic phases were combined, washed once with water (50.0mL) and saturated brine (50mL), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and purified (ethyl acetate: petroleum ether = 1:8) to obtain 16c (748mg, yield: 58%).

[0476] LCMS: Rt: 1.893min; MS m / z (ESI): 501.2[M+H].

[0477] 1 H NMR (400MHz, CDCl3) δ7.20(d,J=3.6Hz,1H),6.63(d,J=3.6Hz,1H),5.61(s,2H),5.50-5.46(m,1H),5.12-5.00(m,1H),3.95 -3.62(m,4H),3.56(t,J=8.4Hz,2H),2.31-2.21(m,1H),2.03-1.95(m,1H),1.32(s,9H),0.95(t,J=5.2Hz,2H),0.00(s,9H).

[0478] Step 2: Synthesis of tert-butyl (3S,4R)4-fluoro-3-((2-((1-((R)-1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)piperidine-1-carboxylate (16e)

[0479] Under nitrogen protection at room temperature, Pd2(dba)3 (14.6 mg, 0.016 mmol) and BINAP (9.7 mg, 0.016 mmol) were added to a solution of 16c (81 mg, 0.162 mmol), (R)-1-(1-methylpyrrolidin-3-yl)-1H-pyrazol-4-amine 16d (53.5 mg, 0.324 mmol) and Cs2CO3 (105 mg, 0.324 mmol) in 1,4-dioxane (5.0 mL), and the reaction system was heated to 100°C and stirred for 16 hours, then cooled to room temperature. The filter cake was filtered and washed with 50.0 mL of tetrahydrofuran, and the solution was concentrated under reduced pressure to obtain the crude product purified to give 16e (67.6 mg, yield: 66.2%).

[0480] LCMS: Rt: 1.420min; MS m / z(ESI): 631.8[M+H].

[0481] Step 3: Synthesis of (4-((3S,4R)-4-fluoropiperidin-3-yl)oxy)-2-((1-((R)-1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methanol (16f)

[0482] At room temperature, TFA (8.0 mL) was added to a solution of 16e (595.8 mg, 1 mmol) in dichloromethane (4.0 mL), stirred at room temperature for 2 hours, concentrated under reduced pressure, added dichloromethane (12.0 mL), and concentrated under reduced pressure to remove excess trifluoroacetic acid. The crude product 16f was directly used in the next step reaction.

[0483] LCMS: Rt: 0.750min; MS m / z (ESI): 431.0 [M+H].

[0484] Step 4: Synthesis of 1-((3S,4R)-4-fluoro-3-((2-((1-((R)-1-methylpyrrolidin-3-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)piperidin-1-yl)prop-2-en-1-one (016)

[0485] The crude product 16f (0.11 mmol) was dissolved in a mixed solvent of acetone (6.0 ml) and water (4.0 mL), the pH value was adjusted to 10 with potassium carbonate, heated to 35 ° C, and stirred for 16 hours. The reaction solution was cooled to 0 ° C, and then acryloyl chloride (45 mg, 0.48 mmol) was dissolved in acetone (1.0 mL) and added dropwise to the reaction solution. After the addition was completed, the reaction solution was filtered and dried. The crude product was purified by Prep-HPLC (Boston Prime C18150*30mm*5μm; A%: water (containing 0.225% FA); B%: ACN15%-50%) to obtain the target product 016 (15.18 mg, yield: 31%).

[0486] LCMS: Rt: 3.617min; MS m / z (ESI): 455.2[M+H].

[0487] 1 H NMR(400M Hz, DMSO-d6) δ11.32(s,1H),9.02(s,1H),8.01(s,1H),7.65(d,J=10.8Hz,1H),6.95(s ,1H),6.92-6.49(m,1H),6.18(d,J=14.4Hz,1H),6.10-5.93(m,1H),5.73-5.41(m,2H) ,5.25-5.12(m,2H),4.09-3.95(m,2H),3.86-3.76(m,3H),3.68-3.51(m,2H),3.44-3. 31(m,2H),2.97-2.90(m,3H),2.33-2.25(m,1H),2.15-2.04(m,1H),1.98-1.93(m,1H).

[0488] Example 17 Preparation of Compound 017

[0489] 1-((3S,4R)-4-fluoro-3-((2-((1-(oxetan-3-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)piperidin-1-yl)prop-2-en-1-one

[0490]

[0491] Synthesis route and specific synthesis steps:

[0492]

[0493] Step 1: Synthesis of 2-chloro-4-(((3S,4R)-4-fluoropiperidin-3-yl)oxy)-7H-pyrrolo[2,3-d]pyrimidine (17b)

[0494] At 0°C, trifluoroacetic acid (0.5 mL) was added to a dichloromethane solution (5.0 mL) of (3S,4R)-3-((2-chloro-7-((2-trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)-4-fluoropiperidine-1-carboxylic acid tert-butyl ester 17a (180 mg, 0.359 mmol), and the reaction solution was stirred at room temperature for 16 hours, concentrated under reduced pressure, and the obtained crude product was added to a methanol solution (5.0 mL) of DIPEA (1.0 mL), and the reaction solution was stirred at 30°C for 2 hours. The solution was concentrated under reduced pressure and purified (acetonitrile / 0.1% trifluoroacetic acid) to obtain 17b (97.2 mg, yield: 99%).

[0495] LCMS: Rt:0.757min; MS m / z(ESI):271.1[M+H].

[0496] Step 2: Synthesis of 4-(((3S,4R)-4-fluoropiperidin-3-yl)oxy)-N-(1-(oxetanyl-3-yl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine (17d)

[0497] Under nitrogen protection, 17b (97.2 mg, 0.359 mmol) and 1-(oxetane-3-yl)-1H-pyrazole-4-amine 17c, 60.0 mg, 0.431 mmol, were dissolved in 1,4-dioxane (10.0 mL), and then Pd2(dba)3 (32.9 mg, 0.036 mmol), BINAP (22.4 mg, 0.036 mmol) and cesium carbonate (292.4 mg, 0.897 mmol) were added under stirring. The reaction system was heated to 100°C, stirred for 16 hours, cooled to room temperature, filtered, and the filter cake was washed with ethyl acetate (60.0 mL). The filtrate was concentrated under reduced pressure to obtain 17d (45.5 mg, yield: 33.9%).

[0498] LCMS: Rt: 0.490min; MS m / z(ESI): 374.4[M+H].

[0499] Step 3: Synthesis of 1-((3S,4R)-4-fluoro-3-((2-((1-(oxetanyl-3-yl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)oxy)piperidin-1-yl)prop-2-en-1-one (017)

[0500] Under stirring at 0℃, 17d (45.5 mg, 0.122 mmol) was dissolved in a mixture of tetrahydrofuran (5.0 mL) and water (2.0 mL), and potassium phosphate (64.7 mg, 0.305 mmol) was added and stirred. Acryloyl chloride (13.3 mg, 0.146 mmol) was added dropwise to the reaction solution and stirred at room temperature for 2 hours. The reaction solution was filtered, and the filtrate was prepared and purified (Prep-HPLC (Boston Prime C18 150*30mm*5μm; A%: water (containing 0.225% FA); B%: ACN 22%-52%)) to obtain 017 (2.15 mg, yield: 4.2%).

[0501] LCMS: Rt:5.527min; MS m / z(ESI):428.1[M+H].

[0502] 1 H NMR(400MHz,DMSO-d6)δ11.34(s,1H),9.00(s,1H),8.04(s,1H),7.65-7.62( m,1H),6.94(s,1H),6.56-6.17(m,1H),6.12-5.92(m,1H),5.74-5.59(m,1H) ,5.50-5.43(m,2H),5.25-5.12(m,1H),4.92-4.85(m,4H),4.04-4.00(m,1H) ,3.90-3.75(m,2H),3.70-3.62(m,1H),3.47-3.39(m,1H),2.14-1.90(m,2H).

[0503] Example 18 Preparation of Compound 018

[0504] 1-((2S,4R)-4-((5-fluoro-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpyrrolidin-1-yl)prop-2-en-1-one

[0505]

[0506] Synthesis route and specific synthesis steps:

[0507]

[0508] Step 1: Synthesis of tert-butyl (2S,4R)-4-((2-chloro-5-fluoro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpyrrolidine-1-carboxylate (18c)

[0509] 2,4-Dichloro-5-fluoro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine 18a (300 mg, 0.98 mmol) and (2S,4R)-4-amino-2-methylpyrrolidine-1-carboxylic acid tert-butyl ester 18b (197 mg, 0.98 mmol) were added to a solution of DIPEA (287 mg, 2.25 mmol) in isopropanol (10.0 mL) and stirred at 110°C for 16 hours. Water (100.0 mL) was added to terminate the reaction, and the mixture was extracted with ethyl acetate (100.0 mL*2). The organic phases were combined and washed with saturated brine (100.0 mL). The organic phases were concentrated under reduced pressure and purified (ethyl acetate: petroleum ether = 1:10) to obtain 18c (380 mg, yield: 85%).

[0510] LCMS: Rt: 1.997min; MS m / z(ESI): 500.1[M+H].

[0511] 1 H NMR (400MHz, DMSO-d6) δ7.64(s,1H),7.40(s,1H),5.47(d,J=9.6Hz,2H),4.59(s,1H),3.87(s,2H),3.55(t,J=8.0Hz,2H ),3.25(s,1H),2.48-2.45(m,1H),1.87(s,1H),1.48(s,9H),1.32(d,J=6.4Hz,3H),0.89(d,J=8.4Hz,2H),0.00(s,9H).

[0512] Step 2: Synthesis of tert-butyl (2S,4R)-4-((5-fluoro-2-((1-methyl-1H-pyrazol-4-yl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpyrrolidine-1-carboxylate (18e)

[0513] At room temperature under nitrogen protection, Pd2(dba)3 (84.2 mg, 0.092 mmol) and BINAP (57.2 mg, 0.092 mmol) were added to a solution of 18c (480 mg, 0.92 mmol), 1-methyl-1H-pyrazol-4-amine 18d (116 mg, 1.19 mmol) and cesium carbonate (749 mg, 2.3 mmol) in 1,4-dioxane (20.0 mL), and the reaction was heated to 120°C and stirred overnight, cooled to room temperature and filtered. The reaction solution was concentrated under reduced pressure and purified (ethyl acetate: petroleum ether = 1:1) to obtain 18e (480 mg, yield: 93%).

[0514] LCMS: Rt: 1.650min; MS m / z(ESI): 561.2[M+H].

[0515] Step 3: 5-Fluoro-N 2 -(1-methyl-1H-pyrazol-4-yl)-N 4 Synthesis of -((3R,5S)-5-methylpyrrolidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidine-2,4-diamine (18f)

[0516] 18e (480 mg, 0.85 mmol) was added to a dichloromethane solution (6.0 mL) of trifluoroacetic acid (3.0 mL) at room temperature, and the reaction solution was stirred at 0°C for 6 hours. The reaction solution was quenched with saturated sodium bicarbonate (50.0 ml), extracted with ethyl acetate (20.0 mL*2), and the organic phase was concentrated under reduced pressure. The crude product was added to a tetrahydrofuran / water solution (4.0 mL / 1.0 mL) of LiOH (522 mg, 12.7 mmol) without purification, and the reaction solution was stirred at 25°C for 2 hours. The reaction solution was concentrated under reduced pressure and reversed to obtain intermediate 18f (80 mg, yield: 28%).

[0517] LCMS: Rt: 1.07min; MS m / z (ESI): 331.2[M+H].

[0518] Step 4: Synthesis of 1-((2S,4R)-4-((5-fluoro-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpyrrolidin-1-yl)prop-2-en-1-one (018)

[0519] 18f (80 mg, 0.24 mmol) was dissolved in a mixture of tetrahydrofuran (6.0 mL) and water (2.0 mL), potassium phosphate (127.0 mg, 0.60 mmol) was added and stirred, and a solution of acryloyl chloride (30.5 mg, 0.34 mmol) in tetrahydrofuran (0.5 mL) was added dropwise, and stirred at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure and purified by Prep-HPLC (acetonitrile / water / 1‰ TFA) to obtain 018 (15.7 mg, yield: 17.0%).

[0520] LCMS: Rt: 6.327min; MS m / z(ESI): 385.1[M+H].

[0521] 1H NMR (400MHz, DMSO-d6) δ10.69(s,1H),8.62(s,1H),7.80(s,1H),7.45(s,1H),6.68-6.53(m,3H),6.15(t,J=12.0Hz,1H),5.68-5.63(m, 1H),4.65-4.58(m,1H),4.22-4.05(m,2H),3.74(s,3H),3.48-3.25(m,1H),2.55-2.42(m,1H),1.94-1.83(m,1H),1.29(d,J=6.0Hz,3H).

[0522] Biological activity and related properties test examples

[0523] Test Example 1: BTK kinase activity inhibition experiment

[0524] Experimental principle: After BTK kinase is incubated with the compound, it reacts with the substrate under the action of ATP. The ADP produced by the reaction is quantified using Promega's ADP-GLO detection kit to reflect the enzyme activity.

[0525] Experimental instruments: Echo650 pipetting system from Labcyte; Envision microplate reader from Perkin Elmer; 5810 centrifuge from Eppendorf.

[0526] Experimental Materials:

[0527] Reagents brand Part Number Tris hydrochloride solution Sigma T2663 BRIJ 35 detergent(10%) Merck 203728 <![CDATA[MgCl2 solution]]> Sigma M1028 ADP-Glo ​​Kinase Assay Kit Promega V9102 BTK Carna bioscience 08-180 Poly(4:1Glu,Tyr) Sigma P0275 384-well plate PerkinElmer 6007290

[0528] Experimental method: Use the Echo pipetting system to transfer the test compound to a 384-well plate, add 2 μL / well of BTK, and incubate for 30 minutes. Then add 3 μL / well of a mixed solution of substrate Poly (4:1Glu, Tyr) and ATP to start the enzyme reaction. The final concentration of the compound starts at 3 μM, 300 nM, or 100 nM, respectively, and is diluted 3 times. The final concentration of the enzyme in the reaction is 1.7 ng / well, the final concentration of ATP is 36 μM, and the final concentration of the substrate is 0.1 mg / mL. After 1 hour of reaction, add 5 μL / well ADP-GLO reagent and incubate for 40 minutes. Then add 10 μL / well kinase reaction detection reagent and incubate for 30 minutes. Use an Envision microplate reader to read the fluorescence signal and calculate the inhibition rate and half inhibitory concentration (IC 50 ).

[0529] The biological activity of the compounds of the present invention was determined by the above test, and the IC 50 The values ​​are shown in Table 1 below.

[0530] Table 1 IC of the Example compounds for inhibition of BTK kinase activity 50

[0531] Example Compound No. <![CDATA[IC 50 (nM)]]> Example Compound No. <![CDATA[IC 50 (nM)]]> 001 6.53 010 3.92 002 7.41 011 5.35 003 3.26 012 9.07 004 3.83 013 4.00 005 4.83 014 0.97 006 6.60 015 2.14 007 1.77 016 1.90 008 2.48 017 3.63 009 3.43 018 1.39

[0532] Test Example 2: JAK3 kinase activity inhibition experiment

[0533] Experimental principle: After JAK3 kinase is incubated with the compound, it reacts with the substrate under the action of ATP. The ADP produced by the reaction is quantified using Promega's ADP-GLO detection kit to reflect the enzyme activity.

[0534] Experimental instruments:

[0535] Labcyte Echo650 Liquid Handling System

[0536] Perkin Elmer Envision Microplate Reader

[0537] Eppendorf 5810 centrifuge.

[0538] Experimental Materials:

[0539] Reagents brand Part Number Tris hydrochloride solution Sigma T2663 BRIJ 35 detergent(10%) Merck 203728 <![CDATA[MgCl2 solution]]> Sigma M1028 ADP-Glo ​​Kinase Assay Kit Promega V9102 JAK3 Carna bioscience 08-046 Poly(4:1Glu,Tyr) Sigma P0275 384-well plate PerkinElmer 6007290

[0540] Experimental methods:

[0541] Use the Echo pipetting system to transfer the test compound to a 384-well plate, add 2 μL / well of JAK3, and incubate for 30 minutes. Then add 3 μL / well of a mixed solution of substrate Poly (4:1Glu, Tyr) and ATP to start the enzyme reaction. The final concentration of the compound starts at 3 μM, 300 nM, or 100 nM, respectively, and is diluted 3 times. The final concentration of the enzyme in the reaction is 1.9 ng / well, the final concentration of ATP is 36 μM, and the final concentration of the substrate is 0.1 mg / mL. After 1 hour of reaction, add 5 μL / well ADP-GLO reagent and incubate for 40 minutes. Then add 10 μL / well kinase reaction detection reagent and incubate for 30 minutes. Use an Envision microplate reader to read the fluorescence signal and calculate the inhibition rate and half inhibitory concentration (IC 50 ).

[0542] The biological activity of the compounds of the present invention was determined by the above test, and the IC 50 The values ​​are shown in Table 2 below.

[0543] Table 2 IC of the compounds of the present invention for inhibition of JAK3 kinase activity 50

[0544] Example Compound No. <![CDATA[IC 50 (nM)]]> Example Compound No. <![CDATA[IC 50 (nM)]]> 001 1.66 010 1.27 002 0.61 011 7.23 003 0.55 012 1.74

[0545] 004 1.36 013 0.58 005 1.41 014 0.51 006 4.74 015 0.25 007 1.48 016 0.89 008 3.04 017 0.84 009 1.81 018 0.46

[0546] Test Example 3: Inhibitory effect on BTK phosphorylation in Ramos cells

[0547] Experimental principle: After incubating Ramos cells with compounds and stimulants, the BTK phosphorylation detection kit of Cisbio is used to detect the transfer of fluorescence energy by homogeneous time-resolved fluorescence (HTRF) method to reflect the inhibitory effect on phosphorylation.

[0548] Experimental instruments:

[0549] instrument brand model Biological Safety Cabinets ESCO CLASSⅡ BSC Centrifuge Eppendorf 5810 <![CDATA[CO2 Incubator]]> ESCO CCL-170B-8 Cell Counter CountStar IC1000 Envision PerkinElmer /

[0550] Experimental Materials:

[0551] Reagents Provider Part Number RPMI-1640 medium GIBCO A10491-01 Inactivated fetal bovine serum (HI-FBS) GIBCO 10100147 HBSS solution GIBCO 14025-092 Anti-human IgM antibody Jackson Immuno 109-006-129 BTK phospho-Y223 HTRF Detection Kit Cisbio 63ADK017PEH 384-well plate PerkinElmer 6007680 Ramos ATCC CRL-1596

[0552] Experimental methods:

[0553] The test compound was transferred to a 384-well plate using the Echo pipetting system and the density of Ramos cells was adjusted to 1×10 7 cells / mL, add 10μL / well cell suspension, and incubate in an incubator at 37°C, 5% CO2 for 1 hour. Then add 5μL / well of stimulator anti-human IgM antibody, the final concentration of the stimulator is 10μg / mL, and incubate for 10 minutes. The final concentration of the compound starts at 1μM and is diluted 4 times. Add 5μL / well of cell lysate and incubate at room temperature for 30 minutes. The phosphorylation degree of BTK was detected using the BTK phospho-Y223 kit from Cisbio, and finally the fluorescence signal at 665nm and 615nm of the emission light was read on the Envision microplate reader to calculate the inhibition rate and half inhibitory concentration (IC 50 ).

[0554] The biological activity of the compounds of the present invention was determined by the above test, and the IC 50 The values ​​are shown in Table 3 below.

[0555] Table 3 Inhibitory activity of the compounds of the present invention on BTK phosphorylation in Ramos cells

[0556] Example Compound No. <![CDATA[IC 50 (nM)]]> Example Compound No. <![CDATA[IC 50 (nM)]]> 001 23.76 010 53.49

[0557] 002 62.24 011 5.04 003 2.63 012 6.15 004 5.07 013 10.85 005 10.90 014 2.02 006 11.34 015 17.89 007 2.39 016 4.23 008 5.78 017 9.66 009 5.73 018 4.01

[0558] Test Example 4: Inhibitory effect on STAT5 phosphorylation in CTLL-2 cells

[0559] Experimental principle: This experiment is to evaluate the effect of compounds on the phosphorylation of STAT5, a downstream substrate of JAK3. After incubating CTLL-2 cells with compounds and stimulants, the p-STAT5 (Tyr694 / 699) detection kit of Perkin Elmer is used to detect the fluorescence energy transfer between donor microbeads and acceptor microbeads by time-resolved fluorescence method, thereby reflecting the inhibitory effect on phosphorylation.

[0560] Experimental instruments:

[0561] instrument brand model Biological Safety Cabinets Thermo Scientific 1300 Series A2 Centrifuge Eppendorf 5702 <![CDATA[CO2 Incubator]]> Thermo Scientific 371 Cell Counter Invitrogen C10281 Envision PerkinElmer / Echo Labcyte 655

[0562] Experimental Materials:

[0563] Reagents brand Part Number Fetal bovine serum (FBS) GIBCO 10099-141 Dimethyl sulfoxide (DMSO) Sigma D8418-1L IL-2 R&D 402-ML-020 CTLL-2 ATCC TIB-214 AlphaLISA SureFire Ultra p-STAT5 (Tyr694 / 699) Assay Kit PerkinElmer ALSU-PST5-B500

[0564] Experimental methods:

[0565] CTLL-2 cells were seeded in 384-well plates at 1.5 × 10 4 cells / 15μl / well, transfer the compound to a 384-well plate using Echo, and incubate in an incubator at 37°C, 5% CO2 for 30 minutes. Then add 5μL / well of the stimulator IL-2 to a final concentration of 1ng / mL and incubate for 30 minutes. The final concentration of the compound starts at 3μM and is diluted 3 times. Add 5μL / well of cell lysate and incubate at room temperature for 10 minutes. The phosphorylation degree of STAT5 was detected using the AlphaLISA p-STAT5 (Tyr694 / 699) detection kit from Perkin Elmer, and finally the AlphaLISA signal was read on the Envision microplate reader to calculate the inhibition rate and half inhibitory concentration (IC 50 ).

[0566] The biological activity of the compounds of the present invention was determined by the above test, and the IC 50 The values ​​are shown in Table 4 below.

[0567] Table 4 Inhibitory activity of the compounds of the present invention on STAT5 phosphorylation in CTLL-2 cells

[0568] Example Compound No. <![CDATA[IC 50 (nM)]]> Example Compound No. <![CDATA[IC 50 (nM)]]>

[0569] 001 11.31 009 46.85 003 19.85 010 143.82 004 44.57 012 13.29 005 18.03 013 17.20 006 13.92 015 47.08 007 185.27 017 76.98 008 200.28 018 27.20

[0570] Test Example 5: Occupancy of BTK target in mouse spleen

[0571] Experimental principle: This experiment is to evaluate the occupancy of the compound on the BTK target in the mouse spleen. The frozen spleen sample is homogenized and then incubated with a biotin-labeled probe compound. The BTK protein not occupied by the compound binds to the probe, while the BTK protein occupied by the compound cannot bind to the probe. The ELISA method is used to detect the occupancy of the compound on the BTK target, thereby reflecting the occupancy of the compound on the BTK target.

[0572] Experimental instruments:

[0573] instrument brand model Centrifuge Eppendorf 5810 Centrifuge Eppendorf 5430R Tissue grinder Beautiful Wall LD48 Envision PerkinElmer /

[0574] Experimental Materials:

[0575] Reagents brand Part Number Anti-BTK Antibody Cell signal 8547S Streptavidin coated plates R&D Systems CP004 CNX-500 MCE HY-100338 ELISA colorimetric solution R&D Systems DY999 ELISA stop solution R&D Systems DY994 RIPA Lysis Buffer Sigma R0278 Anti-rabbit IgG antibody Cell signal 7074S BCA Protein Assay Kit Pierce 23225

[0576] Experimental methods:

[0577] Using C57BL / 6N female mice, the test compound was prepared in 2% Tween80 / 0.5% methycellulose solution and administered by gavage at a dose of 10 mg / kg. The spleen was taken 0.5h or 24 hours after administration and stored in dry ice. The frozen spleen samples were homogenized and the protein concentration was detected using a BCA kit. The spleen homogenate adjusted to the same protein concentration was incubated with the probe compound CNX-500 for 1 hour, and the final concentration of CNX-500 was 1μM. Then 100μL / well was transferred to the streptavidin-coated plate and incubated overnight. Discard the supernatant and wash, add anti-BTK antibody and incubate for 2 hours. Discard the supernatant and wash, add HRP-labeled Anti-rabbit IgG antibody and incubate for 1 hour. Discard the supernatant and wash, use the color developing solution for 10-15 minutes, and after terminating the reaction, use Envision to read the absorbance value at a wavelength of 450nm. Calculate the occupancy ratio, and the occupancy ratio calculation formula is:

[0578]

[0579] in:

[0580] Signal max Indicates: the signal generated after adding the probe compound to the control sample;

[0581] Signal min Indicates: the signal generated by the control sample without adding the probe compound;

[0582] Signal 待测化合物 Indicates: The signal generated after adding the probe compound to the sample of the compound to be tested.

[0583] The experimental results are shown in Table 5.

[0584] Table 5 Occupancy of BTK target sites in mouse spleen by the compounds of the present invention

[0585]

[0586] Test Example 6: Inhibitory effect on IL-2-induced STAT5 phosphorylation in mouse whole blood

[0587] Experimental principle: This experiment is to evaluate the effect of the compound on the phosphorylation of STAT5, a downstream substrate of JAK3. After oral administration, whole blood was collected from mice, and the stimulator IL-2 was added and incubated for 15 minutes. The STAT5 phosphorylation level in lymphocytes was detected by flow cytometry to reflect the inhibitory effect of the compound on the JAK3 target.

[0588] Experimental instruments:

[0589] instrument brand model Biological Safety Cabinets ESCO CLASSⅡ BSC Centrifuge Eppendorf 5810 <![CDATA[CO2 Incubator]]> ESCO CCL-170B-8 Flow cytometer BD Biosciences Canto II -4℃ Refrigerator Haier HYC-650

[0590] Experimental Materials:

[0591] Reagents brand Part Number AF647 Mouse Anti-pStat5 Antibody BD Biosciences 562076 BV 421 anti-mouse CD8 antibody Biolegend 100738 BV421 Anti-Mouse CD3 Antibody BD Biosciences 740014 FITC Anti-Mouse CD4 Antibody BD Biosciences 553047 IL-2 R&D 402-ML-020 Mouse Fc blocking antibody Biolegend 156603 Lysis fixative BD Biosciences 558049 Cell staining solution Biolegend 420201 96-well plate Corning 3799 Perm Buffer III BD Biosciences 558050

[0592] 96-well deep well plate Axygen P-96-450V-C

[0593] Experimental methods:

[0594] Using C57BL / 6N female mice, the test compound was prepared in a 2% Tween80 / 0.5% methycellulose solution and administered by gavage at a dose of 10 mg / kg. After 0.5 hours of administration, whole blood was collected and placed in a sodium heparin anticoagulation tube. 80 μL / well of whole blood was seeded in a 96-well plate, mouse Fc blocking antibody was added, and then 5 μL / well of detection antibody was added. The detection antibody in different batches of experiments was CD8 antibody or CD3 antibody / CD4 antibody mixture. 10 μL / well stimulator IL-2 was added and incubated for 15 minutes. The final concentration of the stimulator was 200 ng / mL. 60 μL / well of whole blood was transferred to a 96-well deep-well plate, and 350 μL / well of lysis fixative was added and incubated for 10 minutes. After centrifugation and discarding the supernatant, the cells were resuspended in 100 μL / well Perm Buffer III and incubated for 30 minutes. Centrifuge and discard the supernatant, add 50 μL / well of pSTAT5 antibody, and incubate for 30 minutes. After centrifugation and discarding the supernatant, the cells were resuspended in the staining solution, the pSTAT5 signal was detected by flow cytometry, and the inhibition rate was calculated. The inhibition rate calculation formula is:

[0595]

[0596] in:

[0597] Signal max Indicates: the signal generated after adding the stimulator IL-2 to the control sample;

[0598] Signal min Indicates: the signal produced by the control sample without the addition of stimulatory agent IL-2;

[0599] Signal 待测化合物 Represents: the signal generated after adding the stimulator IL-2 to the sample of the test compound.

[0600] The experimental results are shown in Table 6.

[0601] Table 6 Inhibitory effect of the compounds of the present invention on IL-2-induced STAT5 phosphorylation in mouse whole blood

[0602]

[0603] The above is an explanation of the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A compound selected from one of the following structures or a pharmaceutically acceptable salt thereof:

2. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

3. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 2 in the preparation of a medicament for preventing or treating Janus kinase (JAK) and / or Bruton's tyrosine kinase (BTK) related diseases.

4. The use according to claim 3, wherein the Janus kinase (JAK) and / or Bruton's tyrosine kinase (BTK) related disease is selected from tumors or autoimmune diseases.

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