Heteroaromatic ring-containing alkynyl compounds, methods for their preparation and uses thereof

Abstract

The present application relates to a class of heteroaromatic ring-containing alkynyl compounds and preparation methods and uses thereof, the heteroaromatic ring-containing alkynyl compounds are compounds of formula (I), or deuterated compounds, pharmaceutically acceptable salts, solvates, esters, acids, metabolites or prodrugs thereof. The compound of formula (I) has a strong killing effect on tumor cells carrying primary KIT mutations and drug-resistant KIT mutations.

Description

Technical Field

[0001] This invention relates to KIT mutation inhibitors, specifically to a class of heterocyclic alkyne compounds, their preparation methods, and uses. More specifically, this invention relates to a compound capable of inhibiting the proliferation of KIT mutant cell lines, a method for preparing such compounds, and their pharmaceutical uses for KIT-related diseases. Background Technology

[0002] KIT (also known as CD117 or stem cell factor receptor) is a 145 kDa transmembrane receptor tyrosine kinase (RTK). KIT belongs to type III RTKs and consists of a cytoplasmic region containing five immunoglobulin-like domains (D1–D5), one transmembrane domain, and one cytoplasmic region containing a juxtamembrane domain (JMD) and a tyrosine kinase (TK) domain. Kinases in the same family include PDGFRα (platelet-derived growth factor receptor α), PDGFRβ (platelet-derived growth factor receptor β), FLT3 (FMS-like tyrosine kinase 3), and CSF-1R (colony-stimulating factor 1 receptor). Under normal circumstances, the binding of ligand stem cell factor (SCF) to the extracellular domain causes receptor dimerization, leading to autophosphorylation of tyrosine residues in the TK domain of the cytoplasm. This further induces the transduction of downstream signaling pathways (such as PI3K, JAK-STAT, Ras-Erk, Src family kinases, and PLC signaling pathways), and triggers various physiological processes, such as cell proliferation, division, and tissue growth and survival.

[0003] KIT gain-of-function mutations are associated with a variety of diseases, including tumors, inflammation, and autoimmune diseases. Numerous studies have shown that KIT mutations are closely related to the occurrence and development of gastrointestinal stromal tumors (GIST), systemic mastocytosis, mast cell leukemia, a small number of acute myeloid leukemias, gliomas, and lung cancer. Among these, the role of mutated KIT in GIST has been studied most extensively and in-depth, with approximately 85% of GIST cases caused by KIT mutations. GIST is the most common mesenchymal tumor of the gastrointestinal tract, with an incidence of approximately 1 / 100,000 to 2 / 100,000, accounting for 1-3% of all gastrointestinal tumors. About 80% of GIST patients have primary KIT mutations located in the juxtamembranous domain (exon 11) and extracellular domain (exon 9). Among these primary mutations, mutations occurring in the JM region (e.g., V560D) are the most common and can lead to ligand-independent constitutive activation of KIT, thereby inducing GIST (Chen, LL et al. Clin. Cancer Res. 2005, 11, pg. 3668-3677; Mol, CD, et al. J. Biol. Chem. 2004, 279, pg. 31655-31663). Imatinib, as a first-line treatment for GIST, is effective against most GISTs with primary KIT mutations, but 90% of patients eventually develop resistance due to secondary mutations in KIT, leading to tumor recurrence. These resistance mutations primarily occur in the ATP-binding pocket of KIT (exon 13, such as K642E, V654A; exon 14, such as T670I) and the activation loop (A-loop; exon 17, such as D816H, D816V, D820A, N822K, N822H, N822V). Sunitinib and Regorafenib, as second- and third-line drugs for GIST, can only overcome a few Imatinib resistance mutations (such as V654A and T670I mutations occurring in the ATP-binding pocket), and their clinical response rates are low. It has been reported that 50% of patients with imatinib resistance have secondary A-loop mutations (Demetri GD, et al. Lancet. 2006, 368:1329-1338; Nishida T, et al. Int. J. Clin. Oncol. 2009, 14:143-149). Therefore, developing novel KIT inhibitors that can reverse resistance caused by A-loop (exon 17) mutations has significant clinical implications.

[0004] Currently, progress in developing KIT inhibitors effective against drug-resistant GIST is slow, especially for small-molecule KIT inhibitors with potent inhibitory activity against resistance mutations occurring in the A-loop. Ripretinib, approved by the FDA in 2020 for the treatment of GIST, can overcome multiple KIT resistance mutations, including exon 17, providing a new treatment option for drug-resistant GIST patients. However, the efficacy of ribretinib still needs improvement; its use as a second-line treatment for GIST failed due to its inability to outperform sunitinib. Furthermore, ribretinib has serious adverse reactions such as hypertension and abdominal pain, which are related to its poor kinase selectivity, particularly its strong inhibitory activity against VEGFR2. Therefore, developing drugs with stronger activity, better KIT selectivity, and greater effectiveness against drug-resistant KIT mutations is of great significance and urgent need for the effective treatment of GIST.

[0005] CN104211639A, CN108456163A, and CN111662227A all relate to heterocyclic alkynyl compounds, including compounds with a linker chain L of -C(O)NH-, -NHC(O)-, ether chain, or amino chain. While the heterocyclic alkynyl compounds disclosed in WO2015089210A1 include -C(O)NHCH2- as the linker chain, their meta-pyridine ring must have a sulfone imine as the dominant structure, and the intermediate benzene ring must be unsubstituented. Furthermore, their focus is on VEGFR and PDGFR, not on inhibitory activity against KIT mutant cell lines. Summary of the Invention

[0006] Based on CN104211639A, CN108456163A, and CN111662227A, the inventors of this application, in their research on the linker chain L being -C(O)NH-, -NHC(O)-, ether chains, or amino chains, discovered that modifying L to form -C(O)NHC(Ra)(Rb)- significantly enhances the inhibitory activity of the compound against KIT mutant cell lines (as shown in Table 1). Based on Example 1, this application found that this novel core structure exhibits excellent inhibitory activity against KIT mutant cell lines with selectivity.

[0007] Table 1. Effect of structure L on activity

[0008]

[0009] This invention provides a novel class of heterocyclic alkyne-based small molecule kinase inhibitors. These compounds exhibit highly potent inhibitory activity against the proliferation of KIT mutant cell lines (including various mutant forms such as KIT D816V). Furthermore, these compounds demonstrate high selectivity, showing no significant cytotoxicity against non-tumor cells (32D cells) or KIT wild-type cells (NCI-H526, Moe7), and exhibiting weak inhibitory activity against EGFR- and PDGFR-dependent cells, reflecting their high selectivity. In addition, representative compounds show significant in vivo tumor-suppressive effects and low toxicity.

[0010] This invention provides a compound of formula (I), or a deuterated compound thereof, a pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug:

[0011]

[0012] in:

[0013] X is selected from -(C(R) a (R) b )) q -;

[0014] Where R a R b Each is independently selected from hydrogen, deuterium, halogen, C1-4 alkyl; or R attached to the same carbon atom. a R b Together with this carbon atom, it forms a 3-5 membered carbon ring;

[0015] q is selected from 1 and 2;

[0016] R 1 Selected from hydrogen, halogens, and C1-6 alkyl groups;

[0017] Ring M 1 Selected from the following structures:

[0018]

[0019] in:

[0020] A 1 A 2 Each independent CR H4 Or N atoms;

[0021] A 3 For CR H2 Or N atoms;

[0022] A 4 For CR H3 NR H3 Or N atoms;

[0023] Z is selected from unsubstituted or substituted 5-10 membered heteroaryl groups and unsubstituted or substituted 5-10 membered heterocyclic groups; the substitution refers to the above groups being independently substituted by 1-5 R groups. 2 Group substitution; or Z and the carbon atom attached to it, as well as the adjacent A. 1 Or A 2 Together they form substituted or unsubstituted rings W 1 The replacement refers to ring W 1 1-5 R 2 Group substitution; in heteroaryl, heterocyclic, or cyclic W 1 In this process, one or more ring C atoms are optionally replaced by a corresponding number of C (=O) groups, and one or more ring S or N atoms are optionally oxidized to form S-oxides or N-oxides;

[0024] W is an unsubstituted or substituted C6-10 aryl group, an unsubstituted or substituted 5-7 membered heteroaryl group, or an unsubstituted or substituted 5-7 membered heterocyclic group; the substitution refers to being replaced by 1-5 R groups. 2 Group substitution; in aryl, heteroaryl, or heterocyclic groups, one or more ring C atoms are optionally replaced by a corresponding number of C(=O) groups, and one or more ring S or N atoms are optionally oxidized to form S-oxides or N-oxides;

[0025] The R 2 Each group is independently selected from halogen, cyano, C1-6 alkyl, C1-6 heteroalkyl, C1-6 alkoxycarbonyl, C2-6 alkenyl, C2-6 alkynyl, C3-6 cycloalkyl, 3-6 membered heterocyclic alkyl, C4-8 cycloalkenyl, 4-8 membered heterocyclic alkenyl, C6-10 aryl, 5-7 membered heteroaryl, 5-7 membered heterocyclic, -C(=O)(C1-6 alkyl), -C(=O)(C3-6 cycloalkyl), -OR 3 -N(R) 3 2, -(C1-6 alkyl)OR 3 -C(=O)N(R) 3 )2、-(C1-6 alkyl)-C(=O)N(R 3 )2、-SR 3 -S(=O)R 3 -S(=O)2R 3 -N(R) 3 )C(=O)R 3 -(C1-6 alkyl)-N(R) 3 )C(=O)R 3 -N(R) 3 )S(=O)2R 3 -P(=O)(R 3 (R) 3); wherein the C1-6 alkyl, C1-6 heteroalkyl, C1-6 alkoxycarbonyl, C2-6 alkenyl, C2-6 alkynyl, C3-6 cycloalkyl, 3-6 heterocyclic alkyl, 4-8 heterocyclic alkenyl, 4-8 heterocyclic alkenyl, C6-10 aryl, 5-7 heteroaryl, 5-7 heterocyclic are optionally surrounded by 1-5 R 3 Replace; or

[0026] Two Rs 2 It forms substituted or unsubstituted 3-7 membered carbon rings or heterocycles with the atoms attached to it, wherein substitution refers to the presence of 1-5 R atoms. 3 replace;

[0027] The R 3 Each is independently selected from hydroxyl, cyano, amino, halogen, C1-6 alkyl, C1-6 alkoxycarbonyl, C3-C6 cycloalkyl, C1-6 heteroalkyl, C1-6 haloalkyl, hydroxyC1-6 alkyl, -NH(C1-6 alkyl), -NH(C3-6 cycloalkyl), -NHC(=O)(C1-6 alkyl), and cyano-substituted 5-7 membered heterocyclic methyl groups selected from one or more of O, N, and S;

[0028] R H1 R H2 and R H4 Each group is independently selected from hydrogen, halogen, amino, cyano, hydroxyl, -N(R) 4 (R) 5 );

[0029] The R 4 R 5 Each is independently selected from hydrogen, C1-6 alkyl, C1-6 heteroalkyl, C3-6 cycloalkyl, C3-6 heterocycloalkyl, C1-6 alkylacyl, and C3-6 cycloalkylacyl.

[0030] R H3 Selected from hydrogen, unsubstituted or substituted C1-6 alkyl, unsubstituted or substituted C1-6 heteroalkyl, -C(=O)OC1-6 alkyl, -C(=O)OC3-6 cycloalkyl, -C(=O)C1-6 alkyl, -C(=O)C3-6 cycloalkyl, unsubstituted or substituted C3-6 cycloalkyl, unsubstituted or substituted C2-6 alkenyl, unsubstituted or substituted C4-6 cycloalkenyl, unsubstituted or substituted C6-10 aryl, unsubstituted or substituted 5-7 membered heteroaryl, and unsubstituted or substituted 4-7 membered heterocyclic groups; wherein the substitution refers to each being independently substituted by 1-5 R groups. 6 The substituted rings are: in cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic groups, one or more cyclic C atoms are optionally replaced by a corresponding number of C (=O) groups, and one or more cyclic S or N atoms are optionally oxidized to form S-oxides or N-oxides;

[0031] The R 6 Each group is independently selected from halogen, cyano, C1-6 alkyl, C1-6 heteroalkyl, C2-6 alkenyl, C2-6 ynyl, C3-6 cycloalkyl, C4-8 cycloalkenyl, C6-10 aryl, 5-7 membered heteroaryl, 5-7 membered heterocyclic, -C(=O)R 7 OR 7 -N(R) 7 2, -(C1-6 alkyl)OR 7 -C(=O)N(R) 7 )2、-(C1-6 alkyl)-C(=O)N(R 7 )2、-SR 7 -S(=O)R 7 -S(=O)2R 7 -N(R) 7 )C(=O)R 7 -(C1-6 alkyl)-N(R) 7 )C(=O)R 7 -N(R) 7 )S(=O)2(R 7 -P(=O)(R) 7 (R) 7 ); wherein the C1-6 alkyl, C1-6 heteroalkyl, C2-6 alkenyl, C2-6 ynyl, C3-6 cycloalkyl, C4-8 cycloalkenyl, C6-10 aryl, 5-7 heteroaryl, and 5-7 heterocyclic groups are optionally surrounded by 1-5 R groups. 7 Replace; or

[0032] Two Rs 6 Together with the atoms attached thereto, they form substituted or unsubstituted 5-7 membered carbon rings or 5-7 membered heterocycles, wherein substitution refers to the presence of 1-5 R atoms. 7 Group substitution;

[0033] The R 7 Each group is independently selected from halogens, hydroxyl groups, cyano groups, amino groups, C1-6 alkyl groups, C2-6 alkenyl groups, C2-6 alkynyl groups, C3-6 cycloalkyl groups, C1-6 heteroalkyl groups, C4-6 heterocycloalkyl groups, C1-6 haloalkyl groups, hydroxyl C1-6 alkyl groups, NH (C1-6 cycloalkyl groups), -NHC (=O) (C1-6 alkyl groups), C (=O) (C1-6 alkyl groups), and C (=O) (C3-6 cycloalkyl groups).

[0034] Ring M 2 Selected from unsubstituted or substituted C6-10 aryl groups, unsubstituted or substituted 5-7 membered heteroaryl groups, and unsubstituted or substituted 5-7 heterocyclic groups; wherein substitution refers to each being independently replaced by 1-5 R groups. M Replaced;

[0035] The R M Each is independently selected from halogen, cyano, C1-6 heteroalkyl, C2-6 alkenyl, C2-6 ynyl, C3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C4-6 cycloalkenyl, C6-10 aryl, 5-7 membered heteroaryl, -C(=O)(C1-6 alkyl), -C(=O)(C3-6 cycloalkyl), -(C1-6 alkyl)-R 8 -OR 8 -N(R) 8 2, -NH(C1-6 alkyl)-R 8 -O(C1-6 alkyl)-R 8 -C(=O)N(R) 8 )2、-SR 8 -S(=O)R 8 -S(=O)2R 8 The C1-6 alkyl, C3-6 cycloalkyl, 3-6 heterocyclic alkyl, C4-6 cycloalkenyl, 4-6 heterocyclic alkenyl, C6-10 aryl, and 5-7 heteroaryl groups are each optionally independently bound by 1-5 R groups. 8 Replace; or

[0036] Two Rs M Together with the atoms attached to it, it forms a substituted or unsubstituted 3-7 membered carbon ring or heterocycle, wherein substitution refers to being formed by 1-5 R atoms. 8 Group substitution;

[0037] The R 8 Each is independently selected from hydroxyl, cyano, amino, halogen, -NH (C1-6 alkyl), C1-6 alkyl, C1-6 heteroalkyl, C1-6 haloalkyl, C1-6 hydroxyalkyl, C3-6 cycloalkyl, and C3-6 heterocycloalkyl.

[0038] In a preferred embodiment, the compound has the structure of the following formula (II):

[0039]

[0040] in,

[0041] R 1 Selected from halogens and C1-3 alkyl groups; preferably selected from methyl, F, and Cl;

[0042] R a R b Each element is independently selected from hydrogen, deuterium, halogen, methyl; or R a and R b Together with the carbon atoms connected to them, they form a ternary carbon ring;

[0043] Ring M 1 Selected from the following structures:

[0044]

[0045] Where A 1 A 2 A 3 A 4 Rings W, Z and R H1 The definitions are as described above; and

[0046] Ring M 2 The definition is as described above.

[0047] In a preferred embodiment, ring M 1 Selected from the following structures:

[0048]

[0049] In this context, "C,N" on the ring atom indicates that the atom is CH or N.

[0050] R 2 R H1 R H2 and R H3 The definitions of each are as described above.

[0051] In a preferred embodiment,

[0052] Ring M 2 Selected from unsubstituted or substituted phenyl, unsubstituted or substituted thiophene, unsubstituted or substituted pyrazolyl, unsubstituted or substituted thiazolyl, unsubstituted or substituted pyridyl, unsubstituted or substituted oxazolyl, unsubstituted or substituted isoxazolyl, unsubstituted or substituted benzothiophene; wherein the substitution refers to each being independently substituted by 1-5 R... M Replaced;

[0053] The R M Each is independently selected from halogen, hydroxyl, amino, cyano, C1-6 alkyl, C1-6 heteroalkyl, C2-6 alkenyl, C2-6 ynyl, C3-6 cycloalkyl, 3-6 membered heterocyclic alkyl, C4-6 cycloalkenyl, 4-6 membered heterocyclic alkenyl, C6-10 aryl, 5-7 membered heteroaryl, -C(=O)(C1-6 alkyl), -C(=O)(C3-6 cycloalkyl), -(C1-6 alkyl)-R 8 -OR 8 -N(R) 8 2, -NH(C1-6 alkyl)-R 8 -O(C1-6 alkyl)-R 8 -C(=O)N(R) 8 )2、-SR 8 -S(=O)R 8-S(=O)2R 8 The C1-6 alkyl, C3-6 cycloalkyl, 3-6 heterocyclic alkyl, C4-6 cycloalkenyl, 4-6 heterocyclic alkenyl, C6-10 aryl, and 5-7 heteroaryl groups are each optionally independently bound by 1-5 R groups. 8 Replace; or

[0054] Two Rs M It forms substituted or unsubstituted 3-7 membered carbon rings or heterocycles with the atoms attached to it, wherein substitution refers to the presence of 1-5 R atoms. 8 Group substitution;

[0055] The R 8 Each is independently selected from hydroxyl, cyano, amino, halogen, -NH (C1-6 alkyl), C1-6 alkyl, C1-6 heteroalkyl, C1-6 haloalkyl, C1-6 hydroxyalkyl, C3-6 cycloalkyl, and C3-6 heterocycloalkyl.

[0056] In this invention,

[0057] The alkyl group is a saturated aliphatic straight-chain or branched alkyl group having 1-8 carbon atoms; typical alkyl groups include, but are not limited to: methyl, ethyl, propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, tert-butyl, etc.

[0058] Each of the halogens is independently selected from F, Cl, Br, and I;

[0059] The haloalkyl group is an alkyl group in which at least one hydrogen atom is replaced by a halogen atom. In some embodiments, if two or more hydrogen atoms are replaced by halogen atoms, the halogen atoms may be the same or different from each other;

[0060] The heteroalkyl group is an alkyl group in which at least one skeletal carbon atom is replaced by a heteroatom (N, O, S). Examples of heteroalkyl groups include alkoxy, alkylamino, alkylthio, etc. In some embodiments, if two or more carbon atoms are replaced by heteroatoms, the heteroatoms may be the same or different from each other.

[0061] The cycloalkyl group is a saturated or partially unsaturated 3-10 member monocyclic or polycyclic alicyclic group, and can be a monovalent group or a bivalent group (i.e., a cycloalkylene group);

[0062] The heterocyclic alkyl group is a saturated 3-10 member monocyclic or polycyclic alicyclic heterocycle containing one or more heteroatoms selected from N, O, and S, and can be a monovalent group or a bivalent group (i.e., a heterocyclic alkylene group).

[0063] The aryl group refers to an aromatic ring in which each constituent atom is a carbon atom, including monocyclic or fused-ring polycyclic rings, and can be a monovalent group or a divalent group (i.e., arylene). In this invention, the aryl ring preferably has 5-10 carbon atoms, and more preferably an aryl group having 5-7 carbon atoms.

[0064] The heteroaryl group is an aromatic group containing one or more heteroatoms selected from N, O, and S on the ring. Depending on the structure, the heteroaryl group can be a monovalent group or a bivalent group (i.e., a heteroaryl group).

[0065] The heterocyclic group is monocyclic or polycyclic, and at least one of them is a non-aromatic cyclic group containing one or more heteroatoms selected from N, O, and S. Depending on the structure, the heterocyclic group can be a monovalent group or a bivalent group (i.e., a hypoheterocyclic group).

[0066] Preferably, the compound of formula (I) is selected from the following compounds:

[0067]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

[0074]

[0075]

[0076] The present invention also provides a method for preparing the above-mentioned compound (I), comprising the following steps: a coupling reaction of compound (A) and compound (B) to obtain compound (I).

[0077]

[0078] Among them, ring M 1 Ring M 2 X and R 1 The definitions are as described above; TMS is -Si(CH3)3;

[0079] Preferably, the coupling reaction is carried out in the presence of a base in the presence of a palladium metal catalyst and a copper metal catalyst, wherein the palladium metal catalyst comprises one or more of Pd(PPh3)2Cl2, Pd(OAc)2, and Pd(PPh3)4; preferably, the copper metal catalyst comprises CuI and / or CuCl; preferably, the base comprises CsF, Cs2CO3, KF, K2CO3, NaHCO3, Na2CO3, Et3N, ( i One or more of Pr)2EtN and DMAP;

[0080] Preferably, the coupling reaction is carried out in the presence of a solvent, which includes one or more of acetonitrile, 1,4-dioxane, and DMF.

[0081] More preferably, the method includes the following steps: the compound of formula (A) and the compound of formula (B) are coupled in an acetonitrile solvent in the presence of cesium fluoride, Pd(PPh3)2Cl2, CuI and triethylamine to obtain the compound of formula (I).

[0082] More preferably, the present invention provides a guiding synthetic scheme (as shown in synthetic route I). It should be understood that the reagents / reaction conditions shown in the synthetic scheme can be modified or optimized using general knowledge of organic chemistry to prepare different compounds of the present invention.

[0083]

[0084] Synthetic Route I

[0085] The A 1 A 2 Ring M 2 X, Z, R H1 and R 1 As described above, each synthetic route I, independently, includes the following steps:

[0086] Step 1: Mix compounds I-1, I-2 and Et3N, add palladium metal catalyst and copper metal catalyst, and react under argon protection (e.g. at room temperature) to obtain compound I-3;

[0087] Step 2: Add compounds I-3 and I-4 to a suitable solvent, add palladium metal catalyst and K2CO3, and heat the reaction under argon protection to obtain important intermediate A; in addition, a suitable ligand may also be added.

[0088] Step 3: Compounds I-5 and I-6 undergo a condensation reaction, wherein X is as described above. Compound I-5, HATU, DIPEA, and DMF are mixed (e.g., stirred at room temperature for 30–60 minutes), then compound I-6 is added, and the reaction is carried out (e.g., at room temperature for 6–12 hours) to obtain intermediate B;

[0089] Step 4: Mix intermediates A and B, base and MeCN, add palladium metal catalyst and copper metal catalyst, and react under argon protection (e.g., in the case of I substitution, react at room temperature, while in the case of Br substitution, react at 80°C, reaction time for example 2 to 6 hours) to obtain compound I-7.

[0090] Preferably,

[0091] The palladium metal catalyst mentioned in steps 1 and 4 is Pd(PPh3)2Cl2, and the copper metal catalyst is CuI;

[0092] The solvent in step 2 is one or more of toluene, ethanol, ethylene glycol dimethyl ether, tetrahydrofuran, 1,4-dioxane, and water; the palladium metal catalyst is any one of Pd(PPh3)4, Pd(OAC)2, and (dppf)PdCl2; the base is any one of K2CO3, Cs2CO3, NaHCO3, and Na2CO3; and the ligand is any one of X-Phos and PPh3.

[0093] The base mentioned in step 4 is cesium fluoride and / or triethylamine;

[0094] More preferably, synthetic route I includes the following steps:

[0095] Step 1: Add compound I-1 and Et3N to a round-bottom flask, replace the oxygen with argon, add Pd(PPh3)2Cl2 and CuI, repeat the oxygen removal process, react at room temperature for about 15 minutes, add I-2, and continue the reaction at room temperature for 3 hours. Purify to obtain compound I-3; wherein, the equivalent amounts of compounds I-1, I-2, Pd(PPh3)2Cl2 and CuI can be approximately 1.0, 1.0~1.5, 0.05~0.1, and 0.1~0.2, respectively.

[0096] Step 2: Add compounds I-3, I-4, K2CO3, THF, and water to a round-bottom flask. Replace the oxygen with argon gas. Add Pd(OAc)2 and X-Phos, and repeat the oxygen removal process. Heat to 80°C and react for 4-8 hours. Purify to obtain compound A. The equivalents of I-3, I-4, K2CO3, Pd(OAc)2, and X-Phos can be 1.0, 1.0-1.5, 2.0-3.0, 0.05-0.1, and 0.1-0.2, respectively. The volume ratio of THF / H2O is 4 / 1.

[0097] Step 3: Add compound I-5, HATU, DIPEA and DMF to a round-bottom flask, stir at room temperature for 30 minutes, then add compound I-6 and react at room temperature; the reaction ends after 12 hours, and compound B can be obtained by purification; the equivalents of compounds I-5, I-6, HATU and DIPEA can be approximately 1.0, 1.0-1.2, 1.0-1.5 and 2.0-4.0, respectively.

[0098] Step 4: Add compounds A, B, Et3N, CsF, and MeCN to a round-bottom flask, replace the oxygen with argon, add Pd(PPh3)2Cl2 and CuI, repeat the oxygen removal process, react at room temperature or heated to 80℃ for 3-6 hours, and then purify to obtain compound I-7; the equivalents of compounds A, B, cesium fluoride, Pd(PPh3)2Cl2, CuI, and Et3N can be approximately 1.0, 1.0-1.5, 2.5-3.0, 0.05-0.1, 0.1-0.2, and 2.5-3.0, respectively.

[0099] The present invention also provides a pharmaceutical composition comprising: one or more selected from compounds of formula (I) above, deuterated compounds thereof, pharmaceutically acceptable salts, solvates, esters, acids, metabolites and prodrugs, and pharmaceutically acceptable excipients.

[0100] The present invention also provides the use of the above-described compound of formula (I) or its deuterated compound, pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug or the above-described pharmaceutical composition in the preparation of KIT (mutant) inhibitors.

[0101] The present invention also provides the use of the above-described compound of formula (I) or its deuterated compound, pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug, or the above-described pharmaceutical composition in the preparation of a medicament for treating, preventing or improving one or more diseases or conditions selected from tumors, inflammation, autoimmune diseases and nervous system diseases.

[0102] Preferably, the tumor is a gastrointestinal stromal tumor, particularly a gastrointestinal stromal tumor involving a KIT mutation, and more particularly a gastrointestinal stromal tumor caused by a KIT mutation that is resistant to Imatinib and / or Sunitinib.

[0103] The compound of formula (I) described in this invention has a strong killing effect on tumor cells carrying primary KIT mutations and drug-resistant KIT mutations.

[0104] According to another specific embodiment of the present invention, the compounds containing heterocyclic alkyne structures of the present invention have shown strong inhibitory activity against tumor cell lines carrying different mutant forms of KIT, namely D816V, V559D, V559D-V654A, V559D / Y823D, and V559D / N822K.

[0105] According to another specific embodiment of the present invention, the compound containing a heterocyclic alkyne structure of the present invention exhibits low cytotoxicity to normal 32D cells and the KIT wild-type cell line NCI-H526, demonstrating the advantage of high toxicity selectivity of the compound.

[0106] According to another specific embodiment of the present invention, the compounds containing heterocyclic alkyne structures of the present invention exhibit low inhibitory activity against BCR-ABL, EGFR, and PDGFR-dependent cell lines, demonstrating the advantage of relative selectivity of such compounds.

[0107] According to another specific embodiment of the present invention, the compound containing a heterocyclic alkyne structure of the present invention exhibits significant inhibitory activity against the growth of related tumors in long-term animal pharmacodynamic models, which is significantly superior to the positive control drug Ripretinib.

[0108] According to another specific embodiment of the present invention, at the effective dosage, the animals are in good condition (including no significant weight loss) and there is no animal death. Attached Figure Description

[0109] Figure 1 The figure shows the effects of compounds 77, 85 and Ripretinib on KIT and its downstream signaling pathways in 32D KIT D816V cells.

[0110] Figure 2 The figure shows the effects of compounds 27, 48 and Ripretinib on KIT and its downstream signaling pathways in 32D KIT V559D cells.

[0111] Figure 3The figure shows the effects of compounds 27, 48 and Ripretinib on KIT and its downstream signaling pathways in 32D KIT V559D-V654A cells.

[0112] Figure 4 The figure shows the therapeutic effects of compounds 77, 85, and Ripretinib on subcutaneous xenografts in 32D KIT D816V nude mice.

[0113] Figure 5 The figure shows the effects of compounds 77, 85, and Ripretinib on the body weight of tumor-bearing (32D KIT D816V) nude mice. Detailed Implementation

[0114] The following provides a detailed description of specific embodiments of the present invention. It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the invention.

[0115] (I) Compound Preparation Examples

[0116] Example 1 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-benzylbenzamide

[0117]

[0118] Step 1: Add 2-amino-5-bromo-3-iodopyridine (1.0 g, 3.35 mmol), trimethylsilylacetylene (427.2 mg, 4.35 mmol), and Et3N (50 mL) to a round-bottom flask. Replace the oxygen with argon. Add Pd(PPh3)2Cl2 (117.4 mg, 0.17 mmol) and CuI (63.7 mg, 0.33 mmol). Repeat the deoxygenation process and react at room temperature for 6 hours. After the reaction is complete, add 50 mL of ethyl acetate to dilute the reaction solution. Filter to obtain 900 mg of the product 2-amino-5-bromo-3-trimethylacetylpyridine (yield: 99.9%).

[0119] Step 2: Add 2-amino-5-bromo-3-trimethylethynylpyridine (300 mg, 1.11 mmol), 1-methylpyrazole-4-boronic acid pinacol ester (348 mg, 1.67 mmol), K2CO3 (385 mg, 2.79 mmol), and THF / H2O (8 / 2 mL) to a round-bottom flask, replace the oxygen with argon, add Pd(OAc)2 (25 mg, 0.11 mmol) and X-Phos (106 mg, 0.22 mmol), and heat to 70 °C for 4 hours. After the reaction was completed, the reaction solution was extracted with ethyl acetate (15 mL × 3) and water (10 mL). The organic phase was washed with saturated NaCl aqueous solution (10 mL × 3), dried over anhydrous Na2SO4, and the solvent was evaporated under reduced pressure. Column chromatography was used to separate 245.0 mg of 5-(1-methyl-1H-pyrazole-4-yl)-3-((trimethylsilyl)ethynyl)-2-aminopyridine (yield: 81.31%).

[0120] Step 3: Add 4-methyl-3-iodobenzoic acid (2.0 g, 7.63 mmol), HATU (3.8 g, 9.92 mmol), DIPEA (2.5 g, 19.08 mmol), and DMF (40 mL) to a round-bottom flask. Stir at room temperature for 30 minutes, then add benzylamine (818 mg, 7.63 mmol) and react at room temperature for 12 hours. After the reaction is complete, extract the reaction solution with ethyl acetate (50 mL × 3) and water (40 mL). Wash the organic phase with tap water (30 mL × 3) and saturated NaCl solution (30 mL × 3), respectively. Dry with anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 2.5 g of N-benzyl-3-iodo-4-methylbenzamide (yield: 93%).

[0121] Step 4: Add N-benzyl-3-iodo-4-methylbenzamide (100 mg, 0.28 mmol), 5-(1-methyl-1H-pyrazol-4-yl)-3-((trimethylsilyl)ethynyl)-2-aminopyridine (100 mg, 0.37 mmol), Et3N (86 mg, 0.85 mmol), CsF (108 mg, 0.71 mmol), and MeCN (20 mL) to a round-bottom flask. Replace the oxygen with argon, add Pd(PPh3)2Cl2 (10 mg, 0.014 mmol) and CuI (5.4 mg, 0.028 mmol), and repeat the deoxygenation process. React at room temperature for 3 hours. After the reaction was completed, the reaction solution was extracted with ethyl acetate (30 mL × 3) and water (20 mL). The organic phase was washed with saturated NaCl solution (10 mL × 3), dried with anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product 3-(2-aminopyridyl-5-bromo-3-ethynyl)-4-methyl-N-benzylbenzamide 78 mg (yield: 65%) was obtained by column chromatography.1 H NMR (400MHz, DMSO-d6) δ9.08(t,J=6.0Hz,1H),8.26(d,J=2.3Hz,1H),8.17(d,J=1.9Hz,1H),8.08(s,1H),7.84(d,J=2.4Hz,1H),7.83–7.79(m,2H ),7.43(d,J=8.0Hz,1H),7.35–7.30(m,4H),7.27–7.21(m,1H),6.28(s, 2H),4.48(d,J=5.9Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS422.1(M+1).

[0122] Example 2 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-phenylethylbenzamide

[0123] Except for replacing benzylamine with phenylethylamine, the synthesis method is as described in Example 1.

[0124] 1 H NMR(400MHz, DMSO-d6)δ8.60(t,J=5.8Hz,1H),8.26(d,J=2.4Hz,1H),8.09(d, J=4.0Hz,2H),7.85(d,J=2.3Hz,1H),7.82(s,1H),7.74(dd,J=1.8,8.0Hz,1H) ,7.41(d,J=8.1Hz,1H),7.33–7.23(m,4H),7.20(t,J=7.2Hz,1H),6.28(s,2H) ,3.84(s,3H),3.49(q,J=6.8Hz,2H),2.85(t,J=7.5Hz,2H),2.53(s,3H).LR-MS 436.1(M+1).

[0125] Example 3 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(1-phenylcyclopropyl)benzamide

[0126] Except for replacing benzylamine with 1-phenylcyclopropylamine, the synthesis method is as described in Example 1.

[0127] 1H NMR (400MHz, DMSO-d6) δ9.21(s,1H),8.26(d,J=2.4Hz,1H),8.16(d,J=1.8Hz,1H),8.08(s,1H),7.84(d,J=2.3Hz,1H),7.82(s,1H),7 .79(d,J=7.5Hz,1H),7.42(d,J=8.0Hz,1H),7.27(t,J=7.6Hz,2H),7.22–7.12(m,3H),6.28(s,2H),3.84(s,3H),2.54(s,3H),1.06(br s,4H).LR-MS 448.2(M+1).

[0128] Example 4 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-benzyl-5-fluorobenzamide

[0129] The synthesis method is as described in Example 1, except that 5-fluoro-3-iodobenzoic acid is used instead of 4-methyl-3-iodobenzoic acid.

[0130] 1 H NMR (400MHz, DMSO-d6) δ9.23(t,J=6.0Hz,1H),8.27(d,J=2.3Hz,1H),8.06(s,1H),8.01(s,1H),7.82(d,J=2.3Hz,1H),7.82–7.7 5(m,3H),7.71(d,J=9.1Hz,1H),7.34(d,J=4.5Hz,4H),7.30–7.22(m,1H),6.46(s,2H),4.50(d,J=5.9Hz,2H),3.84(s,3H).LR-MS 426.2(M+1).

[0131] Example 5 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-5-fluoro-N-(thiophen-3-ylmethyl)benzamide

[0132] Except for replacing 4-methyl-3-iodobenzoic acid with 5-fluoro-3-iodobenzoic acid and benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0133] 1H NMR(400MHz,DMSO-d6)δ9.25(t,J=5.9Hz,1H),8.18(d,J=1.3Hz,1H),8.13–8.06(m,1 H),8.01(d,J=1.1Hz,1H),7.96(t,J=2.0Hz,1H),7.75(d,J=1.5Hz,1H),7.62–7.53(m ,1H),7.36(d,J=7.5Hz,1H),7.05(dd,J=1.6,7.5Hz,1H),7.00(d,J=1.5Hz,1H),6.97 (d,J=1.6Hz,1H),6.29(s,2H),4.55(d,J=5.7Hz,2H),3.87(s,3H).LR-MS432.1(M+1).

[0134] Example 6 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-5-fluoro-N-(pyridin-3-ylmethyl)benzamide

[0135] Except for replacing 4-methyl-3-iodobenzoic acid with 5-fluoro-3-iodobenzoic acid and benzylamine with 3-aminomethylpyridine, the synthesis method is as described in Example 1.

[0136] 1 H NMR (400MHz, DMSO-d6) δ9.23(t,J=6.0Hz,1H),8.64(d,J=1.3Hz,1H),8.47(dd,J=1.3,5.0Hz,1H),8.13(d,J=1.3Hz,1H),8.12–8.07(m,2H),7. 90–7.83(m,3H),7.61–7.54(m,1H),7.50(dd,J=5.0,8.0Hz,1H),7.05(d,J=1.5Hz,1H),6.28(s,2H),4.50(d,J=5.8Hz,2H),3.85(s,3H).LR-MS 427.2(M+1).

[0137] Example 7 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-5-fluoro-N-((6-fluoropyridin-3-yl)methyl)benzamide

[0138] Except for replacing 4-methyl-3-iodobenzoic acid with 5-fluoro-3-iodobenzoic acid and benzylamine with 6-fluoro-3-aminomethylpyridine, the synthesis method is as described in Example 1.

[0139] 1H NMR(400MHz, DMSO-d6)δ9.26(t,J=5.9Hz,1H),8.38(d,J=1.4Hz,1H),8.14(d, J=1.3Hz,1H),8.08(dd,J=1.6,8.7Hz,2H),8.04–7.99(m,1H),7.95(t,J=2.0Hz ,1H),7.85(d,J=1.6Hz,1H),7.53(dt,J=2.0,8.8Hz,1H),7.16(t,J=8.0Hz,1H) ,7.06(d,J=1.6Hz,1H),6.27(s,2H),4.50(d,J=5.6Hz,2H),3.83(s,3H).LR-MS 445.4(M+1).

[0140] Example 8 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-(4-chlorobenzyl)-4-methylbenzamide

[0141] Except for replacing benzylamine with 4-chlorobenzylamine, the synthesis method is as described in Example 1.

[0142] 1 H NMR (400MHz, DMSO-d6) δ9.10(t,J=6.3Hz,1H),8.26(d,J=2.2Hz,1H),8.17(s,1H),8.09(s,1H),7.88(d,J=2.3Hz,1H),7.81(d,J=9. 9Hz,2H),7.44(d,J=8.0Hz,1H),7.41–7.32(m,4H),6.38(s,2H),4.46(d,J=6.0Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS456.7(M+1).

[0143] Example 9 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-(2-trifluoromethylbenzyl)-4-methylbenzamide

[0144] Except for replacing benzylamine with 2-trifluoromethylbenzylamine, the synthesis method is as described in Example 1.

[0145] 1H NMR (400MHz, DMSO-d6) δ9.15(t,J=5.7Hz,1H),8.26(d,J=2.2Hz,1H),8.20(s,1H),8.08(s,1H),7.88–7.79(m,3H),7.74(d,J=7.8Hz,1H ),7.66(t,J=7.6Hz,1H),7.54(d,J=7.8Hz,1H),7.52–7.43(m,2H),6.27(s,2H),4.67(d,J=5.6Hz,2H),3.84(s,3H),2.55(s,3H).LR-MS 490.5(M+1).

[0146] Example 10 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-(2-methoxybenzyl)-4-methylbenzamide

[0147] Except for replacing benzylamine with 2-methoxybenzylamine, the synthesis method is as described in Example 1.

[0148] 1 H NMR (400MHz, DMSO-d6) δ8.88(t,J=5.9Hz,1H),8.26(d,J=2.3Hz,1H),8.18(s ,1H),8.08(s,1H),7.86–7.79(m,3H),7.43(d,J=8.0Hz,1H),7.24(t,J=7.8Hz ,1H),7.19(d,J=7.4Hz,1H),7.00(d,J=8.2Hz,1H),6.91(t,J=7.4Hz,1H),6.2 6(s,2H),4.45(d,J=5.7Hz,2H),3.84(s,3H),3.83(s,3H),2.54(s,3H).LR-MS 452.3(M+1).

[0149] Example 11 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-(3-methoxybenzyl)-4-methylbenzamide

[0150] Except for replacing benzylamine with 3-methoxybenzylamine, the synthesis method is as described in Example 1.

[0151] 1H NMR (400MHz, DMSO-d6) δ9.04(d,J=5.9Hz,1H),8.26(d,J=2.2Hz,1H),8.16(s,1H),8.08(s,1H),7.86–7.77(m,3H),7.43(d,J=8.0Hz,1H),7.2 4(t,J=8.0Hz,1H),6.93–6.86(m,2H),6.82(d,J=8.3Hz,1H),6.26(s,2H),4.45(d,J=5.9Hz,2H),3.84(s,3H),3.73(s,3H),2.53(s,3H).LR-MS 452.3(M+1).

[0152] Example 12 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-(3-methoxybenzyl)-4-methylbenzamide

[0153] Except for replacing benzylamine with 4-methoxybenzylamine, the synthesis method is as described in Example 1.

[0154] 1 H NMR (400MHz, DMSO-d6) δ9.01(t,J=6.1Hz,1H),8.27(d,J=2.3Hz,1H),8.17(s,1H),8.10(s,1H),7.90(s,1H),7.84(s,1H),7.81(d,J=8.3Hz,1H),7. 43(d,J=8.0Hz,1H),7.26(d,J=8.3Hz,2H),6.90(d,J=8.3Hz,2H),6.42(s ,2H),4.41(d,J=5.8Hz,2H),3.85(s,3H),3.73(s,3H),2.54(s,3H).LR-MS 452.3(M+1).

[0155] Example 13 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-(4-fluorobenzyl)-4-methylbenzamide

[0156] Except for replacing benzylamine with 4-fluorobenzylamine, the synthesis method is as described in Example 1.

[0157] 1H NMR (400MHz, DMSO-d6) δ9.08(t,J=6.0Hz,1H),8.26(s,1H),8.16(s,1H),8.09(s,1H),7.87(s,1H),7.84–7.78(m,2H),7.43(d ,J=8.0Hz,1H),7.40–7.32(m,2H),7.16(t,J=8.7Hz,2H),6.36(s,2H),4.46(d,J=5.9Hz,2H),3.84(s,4H),2.53(s,3H).LR-MS 440.2(M+1).

[0158] Example 14 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-(2,4-difluorobenzyl)-4-methylbenzamide

[0159] Except for replacing benzylamine with 2,4-difluorobenzylamine, the synthesis method is as described in Example 1.

[0160] 1 H NMR (400MHz, DMSO-d6) δ9.05(t,J=5.8Hz,1H),8.26(d,J=2.3Hz,1H),8.15(s,1H),8.07(s,1H),7.83(d,J=2.2Hz,1H),7.80(d,J=9.5Hz,2H),7 .43(q,J=8.1Hz,2H),7.23(t,J=10.0Hz,1H),7.07(td,J=2.4,8.6Hz,1H),6.26(s,2H),4.48(d,J=5.6Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS 458.1(M+1).

[0161] Example 15 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-(2-fluoro-4-chlorobenzyl)-4-methylbenzamide

[0162] Except for replacing benzylamine with 2-fluoro-4-chlorobenzylamine, the synthesis method is as described in Example 1.

[0163] 1H NMR(400MHz, DMSO-d6)δ9.07(t,J=5.0Hz,1H),8.26(s,1H),8.15(s,1H),8.07(s,1H),7.84–7.77(m,3H),7.46–7 .37(m,3H),7.28(d,J=8.5Hz,1H),6.26(s,2H),4.48(d,J=5.6Hz,2H),3.84(d,J=1.4Hz,3H),2.53(s,3H).LR-MS 474.7(M+1).

[0164] Example 16 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(pyridin-3-ylmethyl)benzamide

[0165] Except for replacing benzylamine with 3-aminomethylpyridine, the synthesis method is as described in Example 1.

[0166] 1 H NMR (400MHz, DMSO-d6) δ9.13(t,J=5.9Hz,1H),8.57(s,1H),8.47(d,J=3.9Hz,1 H),8.26(d,J=1.8Hz,1H),8.15(s,1H),8.07(s,1H),7.84(d,J=2.3Hz,1H),7.8 2–7.77(m,2H),7.74(d,J=8.0Hz,1H),7.43(d,J=8.0Hz,1H),7.37(dd,J=4.8,7 .8Hz,1H),6.27(s,2H),4.50(d,J=5.8Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS 474.7(M+1).LR-MS 423.5(M+1).

[0167] Example 17 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(pyridin-4-ylmethyl)benzamide

[0168] Except for replacing benzylamine with 4-aminomethylpyridine, the synthesis method is as described in Example 1.

[0169] 1H NMR (400MHz, DMSO-d6) δ9.17(t,J=6.1Hz,1H),8.51(d,J=4.8Hz,2H),8.26(d,J=2.3Hz,1H),8.18(s,1H),8.08(s,1H),7.86–7.79(m ,3H),7.45(d,J=8.1Hz,1H),7.32(d,J=5.3Hz,2H),6.28(s,2H),4.50(d,J=5.9Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS423.5(M+1).

[0170] Example 18 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(pyrimidin-5-ylmethyl)benzamide

[0171] Except for replacing benzylamine with 5-aminomethylpyrimidine, the synthesis method is as described in Example 1.

[0172] 1 H NMR (400MHz, DMSO-d6) δ9.17(t,J=5.8Hz,1H),9.10(s,1H),8.79(s,2H),8.26(d,J=2.3Hz,1H),8.15(s,1H),8.07(s,1H),7.95 –7.81(m,2H),7.79(d,J=8.3Hz,1H),7.43(d,J=8.0Hz,1H),6.28(s,2H),4.50(d,J=5.7Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS 424.5(M+1).

[0173] Example 19 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((6-methoxypyridin-3-yl)methyl)-4-methylbenzamide

[0174] Except for replacing benzylamine with 3-aminomethyl-6-methoxypyridine, the synthesis method is as described in Example 1.

[0175] 1H NMR (400MHz, DMSO-d6) δ9.04(t,J=5.7Hz,1H),8.26(d,J=2.4Hz,1H),8.14(s,2H),8.08(s,1H),7.87–7.80(m,2H),7.78(dd,J=1.9,8.0Hz,1H),7.68( dd,J=2.5,8.5Hz,1H),7.42(d,J=8.0Hz,1H),6.80(d,J=8.5Hz,1H),6.27( s,2H),4.40(d,J=5.8Hz,2H),3.84(s,3H),3.82(s,3H),2.53(s,3H).LR-MS 453.2(M+1).

[0176] Example 20 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((6-chloropyridin-3-yl)methyl)-4-methylbenzamide

[0177] Except for replacing benzylamine with 3-aminomethyl-6-chloropyridine, the synthesis method is as described in Example 1.

[0178] 1 H NMR(600MHz,DMSO-d6)δ9.13(t,J=5.9Hz,1H),8.40(d,J=2.5Hz,1H),8.26(d, J=2.4Hz,1H),8.14(d,J=1.9Hz,1H),8.07(s,1H),7.83(d,J=2.4Hz,1H),7.82 –7.80(m,2H),7.79(dd,J=2.0,8.0Hz,1H),7.50(d,J=8.2Hz,1H),7.43(d,J=8 .0Hz,1H),6.25(s,2H),4.49(d,J=5.8Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS 457.1(M+1).

[0179] Example 21 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((6-fluoropyridin-3-yl)methyl)-4-methylbenzamide

[0180] Except for replacing benzylamine with 3-aminomethyl-6-fluoropyridine, the synthesis method is as described in Example 1.

[0181] 1H NMR (400MHz, DMSO-d6) δ9.13(t,J=5.8Hz,1H),8.26(d,J=2.4Hz,1H),8.21(d,J=2.5Hz,1 H),8.15(d,J=1.9Hz,1H),8.08(s,1H),7.94(td,J=2.6,8.2Hz,1H),7.83(d,J=2.4Hz,1H ),7.82(s,1H),7.79(dd,J=2.0,8.0Hz,1H),7.43(d,J=8.1Hz,1H),7.16(dd,J=2.8,8.5H z,1H),6.27(s,2H),4.49(d,J=5.8Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS441.5(M+1).

[0182] Example 22 Preparation of 3-((2-amino-5-(1-cyclopropyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((6-fluoropyridin-3-yl)methyl)-4-methylbenzamide

[0183] Except for replacing benzylamine with 3-aminomethyl-6-fluoropyridine and replacing 1-methyl-1H-pyrazole-4-boronic acid pinacol ester with 1-cyclopropyl-1H-pyrazole-4-boronic acid pinacol ester, the synthesis method is as described in Example 1.

[0184] 1 H NMR(400MHz, DMSO-d6)δ9.15(t,J=5.7Hz,1H),8.49(d,J=1.4Hz,1H),8.31–8.22(m,1H),8.1 2(d,J=2.2Hz,1H),7.98(d,J=1.3Hz,2H),7.82(d,J=1.3Hz,1H),7.80(dd,J=2.0,7.5Hz,1H), 7.30(dd,J=1.2,7.4Hz,1H),7.08(d,J=1.5Hz,1H),6.92(t,J=8.1Hz,1H),6.39(s,2H),4.50( t,J=5.7Hz,2H),2.52(s,3H),2.44–2.37(m,1H),0.85–0.74(m,2H),0.61–0.49(m,2H).LR-MS 467.2(M+1).

[0185] Example 23 Preparation of 3-((2-amino-5-(1-isopropyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((6-fluoropyridin-3-yl)methyl)-4-methylbenzamide

[0186] Except for replacing benzylamine with 3-aminomethyl-6-fluoropyridine and replacing 1-methyl-1H-pyrazole-4-boronic acid pinacol ester with 1-isopropyl-1H-pyrazole-4-boronic acid pinacol ester, the synthesis method is as described in Example 1.

[0187] 1 H NMR (400MHz, DMSO-d6) δ9.13(t,J=5.7Hz,1H),8.45(d,J=1.3Hz,1H),8.23(d,J=1.3Hz ,1H),8.12–8.04(m,3H),7.86(d,J=1.6Hz,1H),7.75(dd,J=2.0,7.5Hz,1H),7.37(dd,J =1.2,7.5Hz,1H),7.14(t,J=8.0Hz,1H),7.06(d,J=1.5Hz,1H),5.05(s,2H),4.62–4.5 2(m,1H),4.52(t,J=5.6Hz,2H),2.54(s,3H),1.30(d,J=6.8Hz,6H).LR-MS469.5(M+1).

[0188] Example 24 Preparation of 3-((2-amino-5-(1-isopropyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((6-fluoropyridin-3-yl)methyl)-4-methylbenzamide

[0189] Except for replacing benzylamine with 3-aminomethyl-6-methylaminopyridine, the synthesis method is as described in Example 1.

[0190] 1 H NMR (400MHz, DMSO-d6) δ8.90(t,J=5.8Hz,1H),8.26(d,J=2.4Hz,1H),8.12(d,J=1.9Hz,1 H),8.08(s,1H),7.95(d,J=2.4Hz,1H),7.83(d,J=2.4Hz,1H),7.82(s,1H),7.77(dd,J=1 .9,7.9Hz,1H),7.41(d,J=8.1Hz,1H),7.37(dd,J=2.4,8.5Hz,1H),6.43–6.37(m,2H),6. 26(s,2H),4.28(d,J=5.7Hz,2H),3.84(s,3H),2.73(d,J=4.8Hz,3H),2.52(s,3H).LR-MS 452.2(M+1).

[0191] Example 25: Preparation of 5N-((6-(1H-pyrazol-1-yl)pyrazol-3-yl)methyl)-3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methylbenzamide

[0192] Except for replacing benzylamine with (6-(1H-pyrazol-1-yl)pyridin-3-yl)methylamine, the synthesis method is as described in Example 1.

[0193] 1 H NMR(400MHz, DMSO-d6)δ9.16(t,J=5.9Hz,1H),8.60(d,J=2.5Hz,1H),8.45(d,J=1.4Hz,1H), 8.26(d,J=2.4Hz,1H),8.16(d,J=1.9Hz,1H),8.07(s,1H),7.95(dd,J=2.2,8.5Hz,1H),7.91 (dd,J=0.8,8.5Hz,1H),7.83(d,J=2.4Hz,1H),7.82–7.79(m,3H),7.44(d,J=8.1Hz,1H),6.5 7(dd,J=1.7,2.6Hz,1H),6.27(s,2H),4.53(d,J=5.8Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS 489.6(M+1).

[0194] Example 26 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-(3-fluorobenzyl)-4-methylbenzamide

[0195] Except for replacing benzylamine with 3-fluorobenzylamine, the synthesis method is as described in Example 1.

[0196] 1 H NMR (400MHz, DMSO-d6) δ9.10(t,J=6.0Hz,1H),8.26(d,J=2.4Hz,1H),8.17(d,J=1.9Hz, 1H),8.07(s,1H),7.83(d,J=2.4Hz,1H),7.83–7.79(m,2H),7.44(d,J=8.1Hz,1H),7.38 (td,J=6.1,7.9Hz,1H),7.17(d,J=7.8Hz,1H),7.16–7.11(m,1H),7.08(td,J=2.7,8.9H z,1H),6.25(s,2H),4.49(d,J=5.9Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS440.5(M+1).

[0197] Example 27 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0198] Except for replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0199] 1 H NMR (400MHz, DMSO-d6) δ9.01(t,J=5.9Hz,1H),8.26(d,J=2.4Hz,1H),8.15(d,J=1.9Hz ,1H),8.08(s,1H),7.84(d,J=2.4Hz,1H),7.82(s,1H),7.80(dd,J=2.0,8.0Hz,1H),7.4 9(dd,J=3.0,4.9Hz,1H),7.42(d,J=8.1Hz,1H),7.34(dd,J=1.2,3.0Hz,1H),7.09(dd, J=1.2,4.9Hz,1H),6.27(s,2H),4.47(d,J=5.8Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS 427.1(M+1).

[0200] Example 28 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-2-ylmethyl)benzamide

[0201] Except for replacing benzylamine with 2-aminomethylthiophene, the synthesis method is as described in Example 1.

[0202] 1 H NMR (400MHz, DMSO-d6) δ8.97(t,J=5.8Hz,1H),8.30(d,J=1.3Hz,1H),8.17(d,J=2.0H z,1H),8.05(d,J=1.3Hz,1H),7.75(d,J=1.6Hz,1H),7.73(dd,J=2.1,7.6Hz,1H),7.4 4(dd,J=1.6,7.5Hz,1H),7.29(dd,J=1.1,7.7Hz,1H),7.12(dd,J=1.7,7.6Hz,1H),7. 06–6.97(m,2H),6.29(s,2H),4.42(d,J=5.8Hz,2H),3.92(s,3H),2.53(s,3H).LR-MS 428.3(M+1).

[0203] Example 29 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiazolyl-4-ylmethyl)benzamide

[0204] Except for replacing benzylamine with 4-aminomethylthiazole, the synthesis method is as described in Example 1.

[0205] 1 H NMR (400MHz, DMSO-d6) δ9.10(t,J=5.9Hz,1H),9.07(d,J=2.0Hz,1H),8.26(d,J=2.4Hz,1H),8.18(d,J=1.9Hz,1H),8.08(s,1H),7.84(d,J=2 .5Hz,1H),7.83–7.80(m,2H),7.50–7.46(m,1H),7.43(d,J=8.1Hz,1H),6.27(s,2H),4.62(d,J=5.7Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS 429.5(M+1).

[0206] Example 30 Preparation of 4-methyl-3-((5-(1-methyl-1H-pyrazol-4-yl)-2-(methylamino)pyridin-3-yl)ethynyl)-N-(pyridin-3-ylmethyl)benzamide

[0207]

[0208] Step 1: Add 2-chloro-5-bromo-3-iodopyridine (1.0 g, 3.14 mmol) and MeNH2 (2.0 M, 5 mL) to the sealing tube, and heat to 80 °C overnight. After the reaction is complete, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 650 mg of 2-methylamino-5-bromo-3-iodopyridine (yield: 66%). 1 H NMR (400MHz, DMSO-d6) δ8.10(d,J=2.2Hz,1H),8.07(d,J=2.2Hz,1H),6.32–6.24(m,1H),2.78(d,J=4.6Hz,3H).LR-MS 313.1(M+1).

[0209] Step 2: Add 2-methylamino-5-bromo-3-iodopyridine (300 mg, 0.96 mmol), trimethylsilylacetylene (122.4 mg, 1.25 mmol), and Et3N (20 mL) to a round-bottom flask. Replace the oxygen with argon. Add Pd(PPh3)2Cl2 (34 mg, 0.048 mmol) and CuI (18 mg, 0.096 mmol). Repeat the deoxygenation process and react at room temperature for 4 hours. After the reaction is complete, add 20 mL of ethyl acetate to dilute the reaction solution. Filter to obtain 260 mg of the product 2-methylamino-5-bromo-3-trimethylacetylpyridine (yield: 96%). 1 H NMR (400MHz, DMSO-d6) δ8.11(d,J=2.5Hz,1H),7.67(d,J=2.4Hz,1H),6.34(q,J=4.3Hz,1H),2.85(d,J=4.7Hz,3H),0.25(s,9H).LR-MS 284.2(M+1).

[0210] Step 3: Add 2-amino-5-bromo-3-trimethylethynylpyridine (200 mg, 0.71 mmol), 1-methylpyrazole-4-boronic acid pinacol ester (294 mg, 1.41 mmol), K2CO3 (244 mg, 1.77 mmol) and THF / H2O (8 / 2 mL) to a round-bottom flask, replace the oxygen with argon, add Pd(OAc)2 (16 mg, 0.071 mmol) and X-Phos (50 mg, 0.11 mmol), and heat to 70 °C for 3 hours. After the reaction was complete, the reaction solution was extracted with ethyl acetate (15 mL × 3) and water (10 mL). The organic phase was washed with saturated NaCl aqueous solution (10 mL × 3), dried over anhydrous Na₂SO₄, and the solvent was evaporated under reduced pressure. Column chromatography yielded 150 mg of 5-(1-methyl-1H-pyrazole-4-yl)-3-((trimethylsilyl)ethynyl)-2-methylaminopyridine (yield: 74.7%). LR-MS 285.4 (M+1).

[0211] Step 4: Add 4-methyl-3-iodobenzoic acid (2.0 g, 7.63 mmol), HATU (3.8 g, 9.92 mmol), DIPEA (2.5 g, 19.08 mmol), and DMF (40 mL) to a round-bottom flask. Stir at room temperature for 30 minutes, then add 3-aminomethylpyridine (825 mg, 7.63 mmol) and react at room temperature for 12 hours. After the reaction is complete, extract the reaction solution with ethyl acetate (50 mL × 3) and water (40 mL). Wash the organic phase with water (30 mL × 3) and saturated NaCl solution (30 mL × 3), respectively. Dry the solution with anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 2.5 g of 3-iodo-4-methyl-N-(pyridin-3-ylmethyl)benzamide (yield: 93%). LR-MS 353.1 (M+1).

[0212] Step 5: Add 3-iodo-4-methyl-N-(pyridin-3-ylmethyl)benzamide (100 mg, 0.28 mmol), 5-(1-methyl-1H-pyrazol-4-yl)-3-((trimethylsilyl)ethynyl)-2-methylaminopyridine (101 mg, 0.35 mmol), Et3N (86 mg, 0.85 mmol), CsF (108 mg, 0.71 mmol), and MeCN (15 mL) to a round-bottom flask. Replace the oxygen with argon. Add Pd(PPh3)2Cl2 (10 mg, 0.014 mmol) and CuI (5.4 mg, 0.028 mmol). Repeat the deoxygenation process and react at room temperature for 3 hours. After the reaction was completed, the reaction solution was extracted with ethyl acetate (20 mL × 3) and water (20 mL). The organic phase was washed with saturated NaCl solution (10 mL × 3), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product 4-methyl-3-((5-(1-methyl-1H-pyrazol-4-yl)-2-(methylamino)pyridin-3-yl)ethynyl)-N-(pyridin-3-ylmethyl)benzamide 90 mg (yield: 72.6%) was obtained by column chromatography. 1H NMR (400MHz, DMSO-d6) δ9.15(t,J=5.9Hz,1H),8.56(d,J=2.2Hz,1H),8.47(dd,J=1.6,4.8Hz,1 H),8.33(d,J=2.4Hz,1H),8.17(d,J=1.9Hz,1H),8.06(s,1H),7.83(d,J=2.3Hz,1H),7.82–7.7 8(m,2H),7.74(dt,J=2.0,8.0Hz,1H),7.44(d,J=8.1Hz,1H),7.37(dd,J=4.7,7.8Hz,1H),6.45 (q,J=4.3Hz,1H),4.50(d,J=5.8Hz,2H),3.84(s,3H),2.91(d,J=4.6Hz,3H),2.53(s,3H).LR-MS 437.5(M+1).

[0213] Example 31 Preparation of 4-methyl-3-((5-(1-methyl-1H-pyrazol-4-yl)-2-(ethylamino)pyridin-3-yl)ethynyl)-N-(pyridin-3-ylmethyl)benzamide

[0214] Except for replacing methylamine with ethylamine, the synthesis method is as described in Example 30.

[0215] 1 H NMR (400MHz, DMSO-d6) δ9.16(t,J=6.0Hz,1H),8.57(s,1H),8.47(s,1H),8.32(d,J=2.4Hz, 1H),8.15(d,J=1.9Hz,1H),8.06(s,1H),7.83(d,J=2.4Hz,1H),7.83–7.78(m,2H),7.74(d,J =8.0Hz,1H),7.44(d,J=8.0Hz,1H),7.37(dd,J=4.7,7.9Hz,1H),6.34(t,J=5.8Hz,1H),4.50 (d,J=5.8Hz,2H),3.84(s,3H),3.50–3.39(m,3H),2.54(s,3H),1.17(t,J=7.0Hz,3H).LR-MS 451.2(M+1).

[0216] Example 32 Preparation of 3-((2-ethylamino)-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((6-fluoropyridin-3-yl)methyl)-4-methylbenzamide

[0217] Except for replacing methylamine with ethylamine and 3-aminomethyl-6-fluoropyridine with 3-aminomethylpyridine, the synthesis method is as described in Example 37.

[0218] 1 H NMR (400MHz, DMSO-d6) δ9.20(t,J=6.0Hz,1H),8.39(d,J=1.3Hz,1H),8.17(dd,J=1.6,3.2Hz,2 H),8.08(d,J=1.3Hz,1H),8.04–7.98(m,1H),7.93(s,1H),7.84(d,J=1.6Hz,1H),7.66(dd,J=2 .0,7.5Hz,1H),7.36(dd,J=1.2,7.6Hz,1H),7.15(t,J=7.9Hz,1H),7.06(d,J=1.6Hz,1H),4.50 (d,J=5.8Hz,2H),3.85(s,3H),3.47(q,J=8.0Hz,2H),2.54(s,3H),1.09(t,J=8.0Hz,3H).LR-MS 469.3(M+1).

[0219] Example 33 Preparation of 3-((2-ethylamino)-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0220] Except for replacing methylamine with ethylamine and 3-aminomethylthiophene with 3-aminomethylpyridine, the synthesis method is as described in Example 30.

[0221] 1 H NMR (400MHz, DMSO-d6) δ9.14(t,J=6.0Hz,1H),8.22(d,J=1.0Hz,1H),8.16(d,J=2.1Hz,1H),8 .00(d,J=1.3Hz,1H),7.93(s,1H),7.75(d,J=1.5Hz,1H),7.70(dd,J=2.0,7.5Hz,1H),7.36(d ,J=8.1Hz,1H),7.28(dd,J=1.2,7.6Hz,1H),7.06–6.99(m,2H),6.96(d,J=1.6Hz,1H),4.55(t ,J=6.0Hz,2H),3.90(s,3H),3.47(q,J=8.0Hz,2H),2.51(s,3H),1.07(t,J=8.0Hz,3H).LR-MS 456.7(M+1).

[0222] Example 34 Preparation of 3-((2-cyclopropylamino)-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0223] Except for replacing methylamine with cyclopropylamine and 3-aminomethylpyridine with 3-aminomethylthiophene, the synthesis method is as described in Example 30.

[0224] 1 H NMR (400MHz, DMSO-d6) δ9.07(t,J=5.8Hz,1H),8.25(d,J=1.3Hz,1H),8.17(d,J=2.0Hz, 1H),8.16(d,J=1.3Hz,1H),7.88(s,1H),7.85(d,J=1.5Hz,1H),7.68(dd,J=2.0,7.5Hz, 1H),7.43–7.32(m,2H),7.09(d,J=1.6Hz,1H),7.03(dd,J=1.6,7.5Hz,1H),7.00(d,J=1 .5Hz,1H),6.28(s,2H),3.93(s,3H),2.57(s,3H),2.29(m,1H),0.75–0.47(m,4H).LR-MS 468.4(M+1).

[0225] Example 35 3-((2-(N-acetamido)-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0226] Compound 27 (80 mg, 0.187 mmol), Et3N (57 mg, 0.561 mmol), and THF (3 mL) were added to a round-bottom flask. Acetyl chloride (26 μL, 0.374 mmol) was added dropwise under ice bath conditions, and the mixture was stirred at room temperature for 3 h. The reaction was quenched with methanol, and the solvent was then evaporated under reduced pressure. The product was purified by column chromatography to give 70 mg of 3-((2-(N-acetamido)-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide (pale yellow solid; yield: 73%). 1HNMR(400MHz,DMSO-d6)δ9.24(t,J=5.9Hz,1H),8.87(d,J=2.3Hz,1H),8.48–8.38 (m,2H),8.14(s,1H),8.01(d,J=1.9Hz,1H),7.85(dd,J=8.0,2.0Hz,1H),7.46(d, J=8.1Hz,1H),7.40(dd,J=5.1,1.3Hz,1H),7.03(d,J=3.5Hz,1H),6.97(dd,J=5.1 ,3.4Hz,1H),4.63(d,J=5.8Hz,2H),3.91(s,3H),2.44(s,3H),2.25(s,6H).LR-MS 512.2(M+1).

[0227] Example 36 Preparation of 3-((2-amino-5-(3,5-dimethylisoxazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0228] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 3,5-dimethylisoxazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0229] 1 H NMR (400MHz, DMSO-d6) δ9.01(t,J=5.9Hz,1H),8.15(d,J=1.9Hz,1H),7.99(d,J= 2.3Hz,1H),7.80(dd,J=8.0,1.9Hz,1H),7.66(d,J=2.3Hz,1H),7.49(dd,J=4.9,3 .0Hz,1H),7.42(d,J=8.1Hz,1H),7.35–7.31(m,1H),7.09(dd,J=4.9,1.3Hz,1H), 6.50(s,2H),4.46(d,J=5.8Hz,2H),2.51(s,3H),2.37(s,3H),2.20(s,3H).LR-MS 443.1(M+1).

[0230] Example 37 Preparation of 3-((2-amino-5-(1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0231] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0232] 1 H NMR (400MHz, DMSO-d6) δ11.42(s,1H),9.07(t,J=5.8Hz,1H),8.20(d,J=1.3Hz, 1H),8.17(d,J=2.0Hz,1H),8.02(d,J=1.3Hz,1H),7.70(dd,J=2.0,7.5Hz,1H),7 .37(d,J=7.3Hz,1H),7.28(dd,J=1.1,7.4Hz,1H),7.23(s,2H),7.02(dd,J=1.6, 7.3Hz,1H),7.01(s,1H),6.26(s,2H),4.55(d,J=5.8Hz,2H),2.52(s,3H).LR-MS 414.5(M+1).

[0233] Example 38 Preparation of 3-((2-amino-5-(3,5-dimethyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0234] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 3,5-dimethyl-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0235] 1 H NMR (400MHz, DMSO-d6) δ11.95(s,1H),8.96(t,J=5.8Hz,1H),8.18(d,J=1.3H z,1H),8.16(d,J=2.0Hz,1H),7.95(d,J=1.1Hz,1H),7.70(dd,J=2.1,7.6Hz,1 H),7.37(d,J=7.4Hz,1H),7.28(dd,J=1.2,7.2Hz,1H),7.07–6.97(m,2H),6.2 9(s,2H),4.42(d,J=5.7Hz,2H),2.53(s,3H),2.39(s,3H),2.36(s,3H).LR-MS 442.3(M+1).

[0236] Example 39 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-5-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0237] Except for replacing 1-methyl-1H-pyrazole-5-boronic acid tinane ester with 1-methyl-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0238] 1 H NMR (400MHz, DMSO-d6) δ9.14(t,J=5.7Hz,1H),8.79(d,J=1.3Hz,1H),8.08(d,J=2.0H z,1H),7.98(d,J=1.1Hz,1H),7.78(dd,J=2.0,7.5Hz,1H),7.64(d,J=7.5Hz,1H),7.3 7(d,J=7.4Hz,1H),7.28(dd,J=1.1,7.4Hz,1H),7.07–6.97(m,2H),6.35(d,J=7.5Hz, 1H),6.29(s,2H),4.43(d,J=5.7Hz,2H),3.88(s,3H),2.54(s,3H).LR-MS428.1(M+1).

[0239] Example 40 Preparation of 3-((2-amino-5-(1-methyl-1H-imidazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0240] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-methyl-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0241] 1 H NMR (400MHz, DMSO-d6) δ8.76(t,J=5.8Hz,1H),8.88(d,J=1.3Hz,1H),8.17(d,J=2.0Hz,1 H),8.05(d,J=1.3Hz,1H),7.83(s,1H),7.70(dd,J=2.0,7.5Hz,1H),7.36(d,J=7.6Hz,1H ),7.33(s,1H),7.28(dd,J=1.1,7.4Hz,1H),7.05(dd,J=1.3,7.5Hz,1H),7.00(d,J=1.5H z,1H),6.31(s,2H),4.41(d,J=5.7Hz,2H),3.69(s,3H),2.53(s,3H).LR-MS428.1(M+1).

[0242] Example 41 Preparation of 3-((2-amino-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0243] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1,3,5-trimethyl-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0244] 1 H NMR (400MHz, DMSO-d6) δ9.12(t,J=5.7Hz,1H),8.18(d,J=1.3Hz,1H),8.09(d ,J=2.0Hz,1H),7.95(d,J=1.1Hz,1H),7.77(dd,J=2.1,7.6Hz,1H),7.37(d,J= 7.4Hz,1H),7.28(dd,J=1.1,7.6Hz,1H),7.06–6.99(m,2H),6.28(s,2H),4.4 2(d,J=5.8Hz,2H),3.84(s,3H),2.54(s,3H),2.52(s,3H),2.38(s,3H).LR-MS 456.6(M+1).

[0245] Example 42 Preparation of 3-((2-amino-5-(1-cyclopropyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0246] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-cyclopropyl-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0247] 1 H NMR(400MHz, DMSO-d6)δ9.04(t,J=6.0Hz,1H),8.12(d,J=2.0Hz,1H),7.98(s,1H),7.97(s, 1H),7.82(d,J=1.3Hz,1H),7.80(dd,J=2.0,7.5Hz,1H),7.30(dd,J=1.2,7.5Hz,1H),7.19( d,J=8.5Hz,1H),7.08(d,J=1.5Hz,1H),6.88(d,J=11.9Hz,4H),4.46(d,J=5.8Hz,2H),2.45 (s,3H),2.41(q,J=7.0Hz,1H),0.79(ddd,J=4.2,6.0,7.2Hz,2H),0.59–0.49(m,2H).LR-MS 454.2(M+1).

[0248] Example 43 Preparation of 3-((2-amino-5-(1-isopropyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0249] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-isopropyl-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0250] 1 H NMR (400MHz, DMSO-d6) δ9.01(t,J=5.8Hz,1H),8.29(d,J=2.4Hz,1H),8.19(s,1H),8.15(d ,J=1.9Hz,1H),7.86(d,J=2.4Hz,1H),7.82(d,J=0.8Hz,1H),7.80(dd,J=8.0,1.9Hz,1H),7 .49(dd,J=4.9,2.9Hz,1H),7.42(d,J=8.0Hz,1H),7.34(dd,J=3.0,1.3Hz,1H),7.09(dd,J= 5.0,1.3Hz,1H),6.24(s,2H),4.51–4.43(m,3H),2.54(s,3H),1.43(d,J=6.6Hz,6H).LR-MS 456.2(M+1).

[0251] Example 44 Preparation of 3-((2-amino-5-(1-(oxecyclobutyl-3-yl)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0252] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-(oxecyclobutyl-3-yl)-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0253] 1H NMR(400MHz,DMSO-d6)δ9.07(t,J=5.8Hz,1H),8.12(d,J=2.0Hz,1H),7.98(s,1H),7.96 (s,1H),7.82(d,J=1.3Hz,1H),7.80(dd,J=2.0,7.5Hz,1H),7.30(dd,J=1.1,7.4Hz,1H) ,7.22–7.16(m,1H),7.08(d,J=1.5Hz,1H),6.88–6.82(m,2H),6.39(s,2H),5.33–5.26( m,2H),5.08–5.01(m,2H),4.97–4.87(m,1H),4.55(d,J=5.7Hz,2H),2.54(s,3H).LR-MS 470.2(M+1).

[0254] Example 45 Preparation of 3-((2-amino-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0255] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-(piperidin-4-yl)-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0256] 1 H NMR(400MHz, DMSO-d6)δ9.10(t,J=5.7Hz,1H),8.24(d,J=1.3Hz,1H),8.17(d,J=2.1Hz,1H),8.13 (d,J=1.3Hz,1H),7.86(d,J=1.5Hz,1H),7.70(dd,J=2.0,7.5Hz,1H),7.42–7.32(m,2H),7.13(d,J =1.3Hz,1H),7.06–6.99(m,2H),6.28(s,2H),4.46(d,J=5.8Hz,2H),4.30–4.18(m,1H),3.17–3.02 (m,2H),2.78–2.62(m,2H),2.53(s,2H),2.44(s,1H),1.87–1.72(m,2H),1.70–1.51(m,2H).LR-MS 497.3(M+1).

[0257] Example 46 Preparation of 3-((2-amino-5-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0258] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-(1-methylpiperidin-4-yl)-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0259] 1 H NMR (400MHz, DMSO-d6) δ9.06(t,J=5.7Hz,1H),8.24(d,J=1.3Hz,1H),8.17(d,J=2.0Hz,1H),8.13(d, J=1.3Hz,1H),7.86(d,J=1.6Hz,1H),7.70(dd,J=2.1,7.6Hz,1H),7.41–7.34(m,2H),7.13(d,J=1.3Hz ,1H),7.03(dd,J=1.6,7.5Hz,1H),7.00(d,J=1.5Hz,1H),6.27(s,2H),4.45(d,J=5.8Hz,2H),4.33–4 .15(m,1H),3.02–2.83(m,2H),2.53(s,3H),2.25(s,3H),2.22–1.87(m,4H),1.76–1.54(m,2H).LR-MS 511.3(M+1).

[0260] Example 47 Preparation of 3-((2-amino-5-(1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0261] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0262] 1H NMR (400MHz, DMSO-d6) δ9.03(t,J=6.0Hz,1H),8.24(d,J=1.3Hz,1H),8.18(d,J=2.0Hz,1H),8.13(d,J=1. 1Hz,1H),7.87(d,J=1.5Hz,1H),7.70(dd,J=2.0,7.5Hz,1H),7.43–7.33(m,2H),7.13(d,J=1.6Hz,1H),7.0 3(dd,J=1.5,7.5Hz,1H),7.00(d,J=1.5Hz,1H),6.29(s,2H),4.45(d,J=5.8Hz,2H),4.12–3.95(m,1H),3. 95–3.79(m,2H),3.63–3.37(m,2H),2.53(s,3H),2.19–1.91(m,2H),1.89–1.66(m,2H).LR-MS498.6(M+1).

[0263] Example 48 Preparation of 3-((2-amino-5-(1-(2-hydroxyethyl)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0264] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-(2-hydroxyethyl)-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0265] 1 H NMR (400MHz, DMSO-d6) δ9.00(t,J=5.8Hz,1H),8.27(d,J=2.4Hz,1H),8.15(d,J=1.9Hz,1H),8. 10(s,1H),7.85(d,J=2.4Hz,1H),7.84(s,1H),7.80(dd,J=1.9,8.1Hz,1H),7.49(dd,J=3.0,5.0 Hz,1H),7.42(d,J=8.0Hz,1H),7.34(s,1H),7.09(d,J=4.4Hz,1H),6.24(s,2H),4.92(t,J=5.4 Hz,1H),4.47(d,J=5.8Hz,2H),4.13(t,J=5.7Hz,2H),3.75(q,J=5.5Hz,2H),2.53(s,3H).LR-MS 458.1(M+1).

[0266] Example 49 Preparation of 3-((2-amino-5-(1-(2-hydroxypropyl)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0267] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-(2-hydroxypropyl)-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0268] 1 H NMR (400MHz, DMSO-d6) δ9.04(t,J=5.9Hz,1H),8.26(d,J=1.3Hz,1H),8.17(d,J=2.0Hz,1H),8.03(d,J =1.3Hz,1H),7.76(d,J=1.5Hz,1H),7.70(dd,J=2.0,7.5Hz,1H),7.37(d,J=7.4Hz,1H),7.28(d,J=8.0H z,1H),7.06–6.98(m,3H),6.28(s,2H),4.50(d,J=4.8Hz,1H),4.42(d,J=5.8Hz,1H),4.21(dd,J=6.9,1 2.4Hz,1H),4.07(dd,J=6.9,12.3Hz,1H),4.01–3.90(m,1H),2.53(s,3H),1.16(d,J=6.8Hz,3H).LR-MS 472.6(M+1).

[0269] Example 50 Preparation of 3-((2-amino-5-(1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0270] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-(2-(dimethylamino)ethyl)-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0271] 1H NMR (400MHz, DMSO-d6) δ9.04(t,J=5.8Hz,1H),8.26(d,J=1.3Hz,1H),8.17(d,J=2.1Hz, 1H),8.04(d,J=1.3Hz,1H),7.77(d,J=1.6Hz,1H),7.70(dd,J=2.0,7.5Hz,1H),7.37(d, J=7.6Hz,1H),7.28(dd,J=1.2,7.3Hz,1H),7.09–6.96(m,3H),4.66(t,J=7.5Hz,2H),6. 26(s,2H),4.44(d,J=5.8Hz,2H),3.00(t,J=7.5Hz,2H),2.52(s,3H),2.37(s,6H).LR-MS 485.5(M+1).

[0272] Example 51 Preparation of 3-((2-amino-5-(1-(2-fluoroethyl)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0273] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-(2-fluoroethyl)-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0274] 1 H NMR(400MHz, DMSO-d6)δ9.06(t,J=5.7Hz,1H),8.25(d,J=1.3Hz,1H),8.17(d,J=2.0Hz, 1H),8.05(d,J=1.1Hz,1H),7.78(d,J=1.5Hz,1H),7.70(dd,J=2.0,7.5Hz,1H),7.37(d, J=7.5Hz,1H),7.28(dd,J=1.2,7.4Hz,1H),7.08(d,J=1.6Hz,1H),7.05–6.97(m,2H),4. 63(m,2H),4.55(s,2H),4.42(d,J=5.6Hz,2H),3.96(t,J=3.0Hz,2H),2.54(s,3H).LR-MS 460.5(M+1).

[0275] Example 52 Preparation of ethyl acetate 2-(4-(6-amino-5-((2-methyl-5-((thiophen-3-ylmethyl)carbamoyl)phenyl)ethynyl)pyridin-3-yl)-1H-pyrazol-1-yl)

[0276] Except for replacing 1-methyl-1H-pyrazole-1-yl) ethyl acetate with 2-(4-boronic acid phenanthyl ester-1H-pyrazole-1-yl) and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0277] 1 H NMR (400MHz, DMSO-d6) δ9.01(t,J=5.8Hz,1H),8.28(d,J=2.4Hz,1H),8.14(d,J=1.9Hz,1H),8.1 2(s,1H),7.90(s,1H),7.86(d,J=2.4Hz,1H),7.79(dd,J=2.0,7.9Hz,1H),7.48(dd,J=3.0,4.9H z,1H),7.42(d,J=8.1Hz,1H),7.36–7.31(m,1H),7.09(dd,J=1.3,4.9Hz,1H),6.27(s,2H),5.05 (s,2H),4.46(d,J=5.8Hz,2H),4.16(q,J=7.1Hz,2H),2.53(s,3H),1.21(t,J=7.1Hz,3H).LR-MS 500.2(M+1).

[0278] Example 53 Preparation of 3-((2-amino-5-(1-(2-dimethylamino)-2-oxoethyl)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0279]

[0280] Add compound 52 (100 mg, 0.20 mmol) and THF / MeOH / H2O (2 / 0.5 / 0.5 mL) to a round-bottom flask, then add LiOH·H2O (21 mg, 0.50 mmol). After reacting at room temperature for 2 hours, evaporate to dryness under reduced pressure and proceed directly to the next step. The solid was redissolved in DMF (2 mL), and HATU (99 mg, 0.26 mmol) and DIPEA (77 mg, 0.60 mmol) were added. After stirring at room temperature for 20 minutes, a tetrahydrofuran solution of dimethylamine (1.0 M, 0.4 mL) was added, and the reaction was continued for 3 hours. The mixture was then extracted with ethyl acetate (10 mL × 3) and water (10 mL). The organic phases were combined and washed with water (10 mL × 3) and saturated NaCl solution (10 mL × 2), respectively. After drying with anhydrous sodium sulfate, the solvent was evaporated under reduced pressure, and column chromatography was used to separate 55 mg of 3-((2-amino-5-(1-(2-dimethylamino)-2-oxoethyl)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide (yield: 55.3%). 1 H NMR (600MHz, chloroform-d) δ8.17(d,J=2.3Hz,1H),7.91(d,J=1.9Hz,1H),7.71–7.68(m,3H),7.66(d,J=2.3Hz,1H),7.32–7.28(m,2H),7.23–7.21(m,1H), 7.10(dd,J=1.3,5.0Hz,1H),6.67(t,J=5.6Hz,1H),5.09(s,2H),4.99(s, 2H),4.64(d,J=5.6Hz,2H),3.09(s,3H),2.99(s,3H),2.53(s,3H).LR-MS 499.5(M+1).

[0281] Example 54 Preparation of 3-((2-amino-5-(1-trifluoromethyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0282] The synthesis method is the same as in Example 1, except that 1-trifluoromethyl-1H-pyrazolyl-4-boronic acid tinane ester is used instead of 1-methyl-1H-pyrazolyl-4-boronic acid tinane ester.

[0283] 1H NMR (400MHz, DMSO-d6) δ9.06(t,J=5.7Hz,1H),8.26(d,J=1.3Hz,1H),8.16(d,J=2.0Hz ,1H),8.05(d,J=1.3Hz,1H),7.82(d,J=1.5Hz,1H),7.71(dd,J=2.0,7.5Hz,1H),7.37(d ,J=7.4Hz,1H),7.28(dd,J=1.2,7.4Hz,1H),7.22(d,J=1.5Hz,1H),7.03(dd,J=1.5,7. 5Hz,1H),7.00(d,J=1.5Hz,1H),6.25(s,2H),4.42(d,J=5.8Hz,2H),2.52(s,3H).LR-MS 482.1(M+1).

[0284] Example 55 Preparation of 3-((2-amino-5-(1-trifluoromethyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((6-fluoropyridin-3-yl)methyl)-4-methylbenzamide

[0285] Except for replacing 1-methyl-1H-pyrazolyl-4-boronic acid tinane ester with 1-trifluoromethyl-1H-pyrazolyl-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethyl-6-fluoropyridine, the synthesis method is as described in Example 1.

[0286] 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=6.1Hz,1H),8.38(d,J=1.3Hz,1H),8.16(d,J=2.0Hz, 1H),8.13(d,J=1.3Hz,1H),8.10(d,J=1.4Hz,1H),8.07–8.00(m,1H),7.92(d,J=1.5Hz, 1H),7.64(dd,J=2.0,7.5Hz,1H),7.37(dd,J=1.2,7.6Hz,1H),7.28(d,J=1.5Hz,1H),7. 15(t,J=8.0Hz,1H),6.27(s,2H),4.50(d,J=5.9Hz,2H),2.55(s,3H).LR-MS495.3(M+1).

[0287] Example 56 Preparation of 3-((2-amino-5-(1-difluoromethyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0288] The synthesis method is the same as in Example 1, except that 1-difluoromethyl-1H-pyrazolyl-4-boronic acid tinane ester is used instead of 1-methyl-1H-pyrazolyl-4-boronic acid tinane ester.

[0289] 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=6.0Hz,1H),8.20(d,J=1.3Hz,1H),8.16(d,J=2.0H z,1H),8.03(d,J=1.1Hz,1H),7.81(t,J=51.4Hz,1H),7.79(d,J=1.5Hz,1H),7.73–7.6 7(m,1H),7.37(d,J=7.4Hz,1H),7.28(dd,J=1.2,7.5Hz,1H),7.13(d,J=1.6Hz,1H),7 .06–6.97(m,2H),6.29(s,2H),4.55(d,J=5.9Hz,2H),2.54(s,3H).LR-MS464.4(M+1).

[0290] Example 57 Preparation of 3-((2-amino-5-(1-(methyl-d3)-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0291] The synthesis method is the same as in Example 1, except that 1-(methyl-d3)-1H-pyrazolyl-4-boronic acid tinane ester is used instead of 1-methyl-1H-pyrazolyl-4-boronic acid tinane ester.

[0292] 1 H NMR (400MHz, DMSO-d6) δ8.98(t,J=5.9Hz,1H),8.12(d,J=2.0Hz,1H),7.98(d,J=1.3Hz,1H),7.97(d,J=1.6Hz,1H),7.82(d,J=1.3Hz,1H),7.80(dd,J =2.0,7.5Hz,1H),7.30(dd,J=1.1,7.5Hz,1H),7.23–7.15(m,1H),7.05(d, J=1.3Hz,1H),6.91–6.84(m,4H),4.55(d,J=5.7Hz,2H),2.45(s,3H).LR-MS 431.5(M+1).

[0293] The synthesis of 1-(methyl-d3)-1H-pyrazolyl-4-boronic acid linalool ester is described below:

[0294]

[0295] Add 1H-pyrazolyl-4-boronic acid cinnarol ester (0.5 g, 2.58 mmol), cesium carbonate (2.5 g, 7.73 mmol), and DMF (10 mL) to a round-bottom flask. Place in an ice bath and add deuterated iodomethane (411 mg, 2.83 mmol) dropwise. React overnight at room temperature. Pour in water (20 mL), extract with ethyl acetate (25 mL × 3), combine the organic phases, wash with water (20 mL × 3) and saturated NaCl solution (20 mL × 2), dry to anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 467 mg of 1-(methyl-d3)-1H-pyrazolyl-4-boronic acid cinnarol ester (yield: 85.9%). LR-MS (ESI) m / z 212.3 (M+1).

[0296] Example 58 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethylene-d2)benzamide

[0297] Except for replacing benzylamine with thiophene-3-ylmethylene-d2-amine, the synthesis method is as described in Example 1.

[0298] 1 H NMR (400MHz, DMSO-d6) δ9.03(s,1H),,8.12(d,J=2.0Hz,1H),7.97(dd,J=1.4,4.1Hz,2H),7.82(d,J=1.3Hz,1H),7.80(dd,J=2.0,7.5Hz,1H ),7.30(d,J=7.5Hz,1H),7.19(d,J=8.5Hz,1H),7.05(d,J=1.3Hz,1H),6.88(m,2H),6.28(s,2H),3.94(s,3H),2.48(s,3H).LR-MS(ESI)m / z 430.3(M+1).

[0299] The preparation method of thiophene-3-ylmethylene-d2-amine is as follows:

[0300]

[0301] LiAlD4 (962 mg, 22.91 mmol) was added to a round-bottom flask, and the gas was completely replaced with argon. Anhydrous diethyl ether (60 mL) was added, and the mixture was kept in an ice bath under argon protection. A solution of 3-cyanothiophene (1.0 g, 9.16 mmol) in anhydrous diethyl ether (10 mL) was added dropwise. After completion, the mixture was heated to room temperature and refluxed overnight. After cooling to room temperature, the mixture was diluted with diethyl ether (60 mL). A 10% NaOH aqueous solution (2 mL) and water (5 mL) were added dropwise under ice bath. The mixture was filtered through diatomaceous earth, and the filter cake was washed with diethyl ether (25 mL × 3). The solvent was evaporated under reduced pressure, and column chromatography was used to separate 750 mg of thiophene-3-ylmethylene-d2-amine (yield: 71.06%). LR-MS (ESI) m / z 116.2 (M+1).

[0302] Example 59 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((6-fluoropyridin-3-yl)methylene-d2)-4-methylbenzamide

[0303] Except for replacing benzylamine with 6-fluoropyridin-3-ylmethylene-d2-amine, the synthesis method is as described in Example 1.

[0304] 1 H NMR (400MHz, DMSO-d6) δ8.49(d,J=1.4Hz,1H),8.30–8.23(m,1H),8.20(s,1H),8.12(d,J=2.0Hz,1H),7.97(dd,J=1.4,4.1Hz,2H),7.82(d,J=1.3Hz ,1H),7.80(dd,J=2.0,7.5Hz,1H),7.30(dd,J=1.1,7.5Hz,1H),7.05(d,J=1.3Hz,1H),6.94–6.87(m,3H),3.94(s,3H),2.45(s,3H).LR-MS(ESI)m / z 443.2(M+1).

[0305] Example 60 Preparation of methyl-4-(6-amino-5-((2-methyl-5-((thiophen-3-ylmethyl)formyl)phenyl)ethynyl)pyridin-3-yl)-1H-pyrazole-1-carboxylic acid ester

[0306] Except for replacing 1-methyl-1H-pyrazole-4-boronic acid tinane ester with 1-(methoxyformyl)-1H-pyrazole-4-boronic acid tinane ester and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0307] 1H NMR (400MHz, DMSO-d6) δ9.02(t,J=5.9Hz,1H),8.81(s,1H),8.46(d,J=2.4Hz,1H),8.37( s,1H),8.17(d,J=1.9Hz,1H),8.09(d,J=2.4Hz,1H),7.80(dd,J=8.0,2.0Hz,1H),7.49(dd ,J=5.0,2.9Hz,1H),7.43(d,J=8.1Hz,1H),7.34(dd,J=3.0,1.2Hz,1H),7.09(dd,J=4.9, 1.3Hz,1H),6.48(s,2H),4.47(d,J=5.8Hz,2H),4.00(s,3H),2.54(s,3H).LR-MS(ESI)m / z 472.2(M+1).

[0308] The preparation method of 1-(methoxyformyl)-1H-pyrazole-4-boronic acid linalool ester is as follows:

[0309]

[0310] Add 1H-pyrazole-4-boronic acid cinnarol ester (2.0 g, 10.31 mmol) and DMF (30 mL) to a round-bottom flask. Add NaH (60% wt, 618 mg, 15.46 mmol) in portions under ice bath conditions. After stirring for 15 minutes, add methyl chloroformate (1.95 g, 20.61 mmol) and stir at room temperature for 5 hours. After the reaction is complete, quench the reaction by slowly adding saturated NH4Cl aqueous solution in an ice bath. Pour the solution into water (20 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with water (20 mL × 3) and saturated NaCl solution (20 mL × 2), dry with anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 640 mg of 1-(methoxyformyl)-1H-pyrazole-4-boronic acid cinnarol ester (yield: 24.6%). 1 ¹H NMR (400MHz, chloroform-d) δ 8.44 (s, 1H), 7.93 (s, 1H), 4.06 (s, 3H), 1.31 (s, 12H). LR-MS (ESI) m / z 253.1 (M+1).

[0311] Example 61 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((5-cyclopropylthiophen-2-yl)methyl)-4-methylbenzamide

[0312] The synthesis method is as described in Example 1, except that (5-cyclopropylthiophen-2-yl)methylamine is used instead of (5-methylthiophen-2-yl)methylamine.

[0313] 1 H NMR (600MHz, DMSO-d6) δ9.08(t,J=5.9Hz,1H),8.26(d,J=2.4Hz,1H),8.13(d,J=1.9Hz,1H),8.07 (s,1H),7.83(d,J=2.4Hz,1H),7.81(d,J=0.8Hz,1H),7.78(dd,J=7.9,1.9Hz,1H),7.42(d,J=8.1 Hz,1H),6.78(d,J=3.5Hz,1H),6.63(dd,J=3.5,0.8Hz,1H),6.25(s,2H),4.52(d,J=5.8Hz,2H),3 .84(s,3H),2.53(s,3H),2.09–2.00(m,1H),0.96–0.90(m,2H),0.63–0.56(m,2H).LR-MS(ESI)m / z 468.3(M+1).

[0314] The preparation method of (5-cyclopropylthiophene-2-yl)methylamine is as follows:

[0315]

[0316] 5-Cyclopropylthiophene-2-carboxaldehyde (1.0 g, 6.57 mmol), NH4OAc (2.53 g, 32.85 mmol), and MeOH (20 mL) were added to a round-bottom flask, followed by NaBH3CN (1.24 g, 19.71 mmol). The mixture was stirred at 50 °C for 12 h. After the reaction was complete, the mixture was poured into a saturated NaHCO3 aqueous solution, and most of the methanol was evaporated under reduced pressure. The mixture was extracted with ethyl acetate (30 mL × 3), and the organic phases were combined. The mixture was washed with saturated NaCl solution (20 mL × 2), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. Column chromatography was used to separate 300 mg of (5-cyclopropylthiophene-2-yl)methylamine (yield: 29.8%). 1 H NMR(400MHz,DMSO-d6)δ7.05(d,J=6.2Hz,1H),6.82(d,J=6.1Hz,1H),5.03(br s,2H),4.30(s,2H),2.29–2.15(m,1H),1.03–0.85(m,4H).LR-MS(ESI)m / z 128.1(M+1).

[0317] Example 62 Preparation of 3-((2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)ethynyl)-N-((5-chlorothiophen-2-yl)methyl)-4-methylbenzamide

[0318] The synthesis method is as described in Example 61, except that 5-chlorothiophene-2-aldehyde is used instead of 5-cyclopropylthiophene-2-aldehyde.

[0319] 1 H NMR (400MHz, DMSO-d6) δ8.83(t,J=8.0Hz,1H),8.18(s,1H),8.12(d,J=2.1Hz,1H ),7.98(d,J=2.0Hz,1H),7.82(d,J=2.0Hz,1H),7.77(dd,J=7.6,1.9Hz,1H),7.3 3(dq,J=7.6,1.0Hz,1H),7.30(s,1H),7.09(s,2H),6.92(d,J=6.4Hz,1H),6.60( d,J=6.4Hz,1H),4.75(d,J=8.0Hz,2H),3.94(s,3H),2.45(s,3H).LR-MS(ESI)m / z 463.0(M+1).

[0320] Example 63 Preparation of 3-((2-amino-5-(4-cyclopropyl-1H-imidazol-1-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0321]

[0322] Step 1: Add 4-methyl-3-iodobenzoic acid (2.0 g, 7.63 mmol), HATU (3.8 g, 9.92 mmol), DIPEA (2.5 g, 19.08 mmol), and DMF (40 mL) to a round-bottom flask. Stir at room temperature for 30 minutes, then add 3-aminomethylthiophene (864 mg, 7.63 mmol) and react at room temperature for 12 hours. After the reaction is complete, extract the reaction solution with ethyl acetate (50 mL × 3) and water (40 mL). Wash the organic phase with water (30 mL × 3) and saturated NaCl aqueous solution (30 mL × 3), respectively. Dry the solution with anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 2.4 g of 3-iodo-4-methyl-N-(thiophene-3-ylmethyl)benzamide (pale yellow solid; yield: 88%).

[0323] Step 2: Add 3-iodo-4-methyl-N-(thiophen-3-ylmethyl)benzamide (100 mg, 0.28 mmol), 5-bromo-3-((trimethylsilyl)ethynyl)-2-aminopyridine (90 mg, 0.34 mmol), Et3N (85 mg, 0.84 mmol), CsF (128 mg, 0.84 mmol), and MeCN (20 mL) to a round-bottom flask. Replace the oxygen with argon. Add Pd(PPh3)2Cl2 (10 mg, 0.014 mmol) and CuI (5.3 mg, 0.028 mmol). Repeat the deoxygenation process and react at room temperature for 3 hours. After the reaction was completed, the reaction solution was extracted with ethyl acetate (30 mL × 3) and water (20 mL). The organic phase was washed with saturated NaCl solution (10 mL × 3), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product 3-((2-amino-5-bromopyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide 85 mg (yellow solid; yield: 71%) was obtained by column chromatography.

[0324] Step 3: Add 3-((2-amino-5-bromopyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide (80 mg, 0.19 mmol), 4-cyclopropylimidazolium (30 mg, 0.28 mmol), CuI (7 mg, 0.038 mmol), Cs₂CO₃ (122 mg, 0.38 mmol), and DMF (2 mL) to a round-bottom flask. React at 120 °C for 30 h under argon protection. Cool to room temperature, add 10 mL of ethyl acetate to dilute the reaction solution, filter, wash the filter cake with ethyl acetate, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 50 mg of the product 3-((2-amino-5-(4-cyclopropyl-1H-imidazol-1-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide (yellow solid; yield: 59%). 1H NMR (400MHz, DMSO-d6) δ8.82(t,J=6.0Hz,1H),8.52(d,J=2.0Hz,1H),8.12(d,J=2.0Hz,1H), 7.80(d,J=1.5Hz,1H),7.77(dd,J=7.6,1.9Hz,1H),7.60(d,J=2.0Hz,1H),7.33(dq,J=7.6,1. 1Hz,1H),7.22(dd,J=4.8,1.8Hz,1H),7.16(d,J=2.2Hz,1H),7.07–7.02(m,2H),6.25(s,2H), 4.55(d,J=5.9Hz,2H),2.45(d,J=0.9Hz,3H),2.14(pd,J=7.9,0.7Hz,1H),1.33–1.11(m,4H).

[0325] Example 64 Preparation of 3-((3-aminoisoquinoline-4-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0326] Except for replacing 5-bromo-3-iodopyridine-2-amine with 4-iodo-isoquinoline-3-amine and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0327] 1 H NMR(400MHz,DMSO-d6)δ9.03(t,J=6.1Hz,1H),8.91(s,1H),8.26(d,J=1.9Hz,1H),7.93 (t,J=8.3Hz,2H),7.80(dd,J=8.0,2.0Hz,1H),7.71–7.66(m,1H),7.50(dd,J=5.0,2.9H z,1H),7.45(d,J=8.1Hz,1H),7.35(d,J=1.4Hz,1H),7.30(dd,J=8.3,7.1Hz,1H),7.11( dd,J=5.0,1.3Hz,1H),6.54(s,2H),4.49(d,J=5.9Hz,2H),2.61(s,3H).LR-MS(ESI)m / z 398.1(M+1).

[0328] Example 65 Preparation of 3-((2-aminopyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0329] Except for replacing 5-bromo-3-iodopyrimidine-2-amine with 5-iodopyrimidine-2-amine and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0330] 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.9Hz,1H),8.45(s,2H),7.99(d,J=1.9Hz,1H),7.78(dd,J=8.0,1.9Hz,1H),7.48(dd,J=4.9,2.9Hz,1H ),7.41(d,J=8.1Hz,1H),7.36–7.31(m,1H),7.19(s,2H),7.08(dd,J=4.9,1.3Hz,1H),4.45(d,J=5.8Hz,2H),2.48(s,3H).LR-MS(ESI)m / z 349.1(M+1).

[0331] Example 66 Preparation of 3-((2-amino-5-(5,5-dimethyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0332] Except for replacing N-methylpyrazole-4-boronic acid pinacol ester with 5,5-dimethyl-5,6-dihydro-4H-pyrrole[1,2-b]pyrazole-3-boronic acid pinacol ester (synthetic method referred to J.Med.Chem.2020,63,24,15564–15590) and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as in Example 1.

[0333] 1 H NMR (400MHz, chloroform-d) δ8.11(d,J=2.3Hz,1H),7.94(d,J=2.0Hz,1H),7.71–7.6 6(m,2H),7.61(d,J=2.3Hz,1H),7.32–7.27(m,2H),7.22–7.19(m,1H),7.08 (dd,J=5.0,1.3Hz,1H),6.80(t,J=5.7Hz,1H),5.07(s,2H),4.63(d,J=5.6H z,2H),3.87(s,2H),2.83(s,2H),2.53(s,3H),1.32(s,6H).LR-MS(ESI)m / z 482.2(M+1).

[0334] Example 67 Preparation of 3-((2-amino-5-(2-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)thiazolyl-4-yl)pyridin-3-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0335]

[0336] Step 1: Except for replacing N-methylpyrazole-4-boronic acid pinacol ester with (2-((tert-butoxyformyl)((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)thiazol-4-boronic acid pinacol ester (synthetic method referred to Eur. J. Med. Chem., 158(2018): 896-916.) and replacing benzylamine with 3-aminomethylthiophene, intermediate 67-IM can be prepared by referring to the synthetic method of Example 1. 1 ¹H NMR (400 MHz, chloroform-d) δ 8.54 (s, 1H), 8.02–7.99 (m, 1H), 7.93 (d, J = 2.0 Hz, 1H), 7.70 (dd, J = 8.0, 1.9 Hz, 1H), 7.34–7.29 (m, 2H), 7.22 (s, 1H), 7.12–7.07 (m, 1H), 7.01 (s, 1H), 6.54 (s, 1H),5.27(s,2H),4.65(d,J=5.5Hz,2H),4.51(s,2H),3.98–3.89(m,2H),3.70(td,J=11. 8,3.3Hz,2H),2.55(s,3H),1.93–1.83(m,4H),1.64(s,9H).LR-MS(ESI)m / z669.2(M+1).

[0337] Step 2: Add intermediate 67-IM (50 mg, 0.075 mmol) and DCM (2 mL) to a round-bottom flask, then add TFA (0.5 mL) and react overnight at room temperature. Pour the reaction solution into an aqueous NaHCO3 solution and extract with DCM. Combine the organic phases, wash with an aqueous NaCl solution, dry under anhydrous Na2SO4, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 40 mg of the product (pale yellow solid; yield: 94%). 1H NMR(400MHz, DMSO-d6)δ9.01(t,J=5.9Hz,1H),8.52(d,J=2.3Hz,1H),8.16(d,J=2.0Hz,1H),8.09–8.01(m,2H),7 .80(dd,J=8.0,2.0Hz,1H),7.49(dd,J=5.0,3.0Hz,1H),7.42(d,J=8.0Hz,1H),7.34(dd,J=2.9,1.2Hz,1H),7.09( dd,J=5.0,1.3Hz,1H),6.99(s,1H),6.44(s,2H),4.47(d,J=5.8Hz,2H),3.92(d,J=11.2Hz,2H),3.66(d,J=6.3Hz ,2H),3.55–3.41(m,2H),2.53(s,3H),1.89(d,J=13.5Hz,2H),1.71(td,J=13.1,12.7,4.4Hz,2H).LR-MS(ESI)m / z 569.2(M+1).

[0338] Example 68 Preparation of 4-methyl-3-((6-(1-methyl-1H-pyrazol-4-yl)imidazol[1,2-b]pyrazin-3-yl)ethynyl)-N-(thiophene-3-ylmethyl)benzamide

[0339] Except for replacing 5-bromo-3-iodopyridin-2-amine with 6-chloro-3-iodoimidazole[1,2-b]pyrazine and benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0340] 1 H NMR (400MHz, DMSO-d6) δ9.11(t,J=6.6Hz,1H),8.47(s,1H),8.24(d,J=9.5Hz,1H),8.18–8.05(m,3H),7.86(d,J=8.0Hz,1H),7.72(d,J= 9.4Hz,1H),7.50(d,J=7.0Hz,2H),7.35(s,1H),7.11(d,J=5.0Hz,1H),4.48(d,J=5.7Hz,2H),3.94(s,3H),2.67(s,3H).LR-MS(ESI)m / z 453.1(M+1).

[0341] Example 69 Preparation of tert-butyl-5-((2-methyl-5-((thiophen-3-ylmethyl)carbamoyl)phenyl)ethynyl)-1H-pyrazole[3,4-b]pyridine-1-carbonate

[0342] Except for replacing 5-bromo-3-iodopyridine-2-amine with tert-butyl-5-bromo-1H-pyrazole[3,4-b]pyridine-1-carbonate and replacing benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0343] 1 H NMR (400MHz, chloroform-d) δ8.88(d,J=2.0Hz,1H),8.23(d,J=2.0Hz,1H),8.19(s,1H),7.94(d,J=2.0Hz,1H),7.71(dd,J=8.0,2.0Hz,1H),7.37–7 .31(m,3H),7.27–7.21(m,15H),7.11(dd,J=4.9,1.3Hz,1H),6.34(s,1H),4.67(d,J=5.5Hz,2H),2.58(s,3H),1.74(s,9H).LR-MS(ESI)m / z 473.2(M+1).

[0344] Example 70 Preparation of 3-((1H-pyrazol[3,4-b]pyridin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0345] Except that compound 79 is replaced by compound 83, the synthesis method is as described in Example 80.

[0346] 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),9.07(t,J=5.9Hz,1H),8.72(d,J=2.0Hz,1H),8 .51(d,J=2.0Hz,1H),8.21(d,J=1.4Hz,1H),8.08(d,J=1.9Hz,1H),7.83(dd,J=8.0,1 .9Hz,1H),7.49(dd,J=5.0,3.0Hz,1H),7.45(d,J=8.1Hz,1H),7.34(dd,J=2.9,1.3Hz ,1H),7.09(dd,J=4.9,1.3Hz,1H),4.47(d,J=5.8Hz,2H),2.55(s,3H).LR-MS(ESI)m / z 373.1(M+1).

[0347] Example 71 Preparation of 3-(imidazolium[1,2-a]pyrimidin-3-ylethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0348] Except for replacing 5-bromo-3-iodopyridin-2-amine with 3-bromoimidazole[1,2-a]pyrimidine and benzylamine with 3-aminomethylthiophene, the synthesis method is as described in Example 1.

[0349] 1 H NMR (400MHz, chloroform-d) δ8.81(dd,J=4.6,1.6Hz,1H),8.21(dd,J=8.0,1.7Hz,1H),8.01(d,J=2.0Hz,1H),7.76(dd,J=8.1,2.0Hz,1H),7.40(dd,J=8.0, 4.6Hz,1H),7.38–7.33(m,2H),7.25(s,2H),7.12(dd,J=4.9,1.3Hz,1H),6.36(t,J=6.5Hz,1H),4.67(d,J=5.5Hz,2H),2.63(s,3H).LR-MS(ESI)m / z 373.2(M+1).

[0350] Example 72 Preparation of 3-((4-amino-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0351]

[0352] Step 1: Add 5-iodo-7H-pyrrolo[2,3-d]pyrimidine-4-amine (1.0 g, 3.85 mmol), K₂CO₃ (1.59 g, 11.54 mmol), and anhydrous DMF (10 mL) to a round-bottom flask, and add iodomethane (0.29 mL, 4.61 mmol) dropwise. React at room temperature for 8 hours. After the reaction is complete, add 2 mL of sodium thiosulfate aqueous solution, stir at room temperature for 15 min, then pour into water, extract with DCM, combine the organic phases, wash with water and NaCl aqueous solution respectively, dry with anhydrous Na₂SO₄, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 600 mg of 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine-4-amine (yield: 60%). Different N-substituted products can be obtained by changing different halogenated products and adjusting reaction conditions (such as base, temperature, etc.).

[0353] Step 2: Add 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine-4-amine (0.5 g, 1.82 mmol), trimethylsilylacetylene (233 mg, 2.37 mmol), Et3N (5 mL), and MeCN (20 mL) to a round-bottom flask. Replace the oxygen with argon. Add Pd(PPh3)2Cl2 (64 mg, 0.091 mmol) and CuI (17.4 mg, 0.091 mmol). Repeat the deoxygenation process and react overnight at room temperature. After the reaction is complete, add 50 mL of ethyl acetate to dilute the reaction solution. Filter, evaporate the solvent under reduced pressure, and separate the product by column chromatography to obtain 420 mg of 7-methyl-5-((trimethylsilyl)acetylene)-7H-pyrrolo[2,3-d]pyrimidine-4-amine (yield: 94.2%).

[0354] Step 3: Add 3-iodo-4-methyl-N-(thiophene-3-ylmethyl)benzamide (90 mg, 0.25 mmol), 7-methyl-5-((trimethylsilyl)ethynyl)-7H-pyrrolo[2,3-d]pyrimidine-4-amine (62 mg, 0.25 mmol), Et3N (76 mg, 0.76 mmol), CsF (115 mg, 0.76 mmol), and MeCN (10 mL) to a round-bottom flask. Replace the oxygen with argon. Add Pd(PPh3)2Cl2 (9 mg, 0.013 mmol) and CuI (5 mg, 0.025 mmol). Repeat the deoxygenation process and react at room temperature for 3 hours. After the reaction was completed, the reaction solution was poured into water and extracted with ethyl acetate. The organic phase was washed with saturated NaCl solution (10 mL × 3), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product 3-((4-amino-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide 60 mg (light yellow solid; yield: 59%) was obtained by column chromatography. 1 H NMR (400MHz, DMSO-d6) δ9.04(t,J=5.7Hz,1H),8.17(s,1H),8.06(s,1H),7.79(d,J=8.0Hz,1H),7.70(s,1H),7.49(s,1H),7.42(d ,J=8.0Hz,1H),7.34(s,1H),7.09(d,J=5.0Hz,1H),6.71(s,2H),4.47(d,J=5.8Hz,2H),3.73(s,3H),2.52(s,3H).LR-MS(ESI)m / z 402.1(M+1).

[0355] Example 73 Preparation of 3-((4-amino-7-(methyl-d3)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0356] Except for replacing iodomethane with deuterated iodomethane, the synthesis method is as described in Example 72.

[0357] 1 H NMR (400MHz, DMSO-d6) δ9.02(t,J=6.0Hz,1H),8.20(s,1H),8.08(s,1H),7.79(d,J=7.8Hz,1H),7.70(s,1H),7.50 (s,1H),7.42(d,J=8.0Hz,1H),7.26-7.19(m,2H),6.63(s,2H),4.45(d,J=5.9Hz,2H),2.53(s,3H).LR-MS(ESI)m / z 405.1(M+1).

[0358] Example 74 Preparation of tert-butyl-4-amino-5-((2-methyl-5-(thiophen-3-ylmethyl)carbamoyl)phenyl)ethynyl)-7H-pyrrolo[2,3-d]pyrimidine-7-carbonate

[0359] The synthesis method is as described in Example 72, except that tert-butyl 4-amino-5-iodo-7H-pyrrole[2,3-d]pyrimidine-7-carbonate is used instead of 5-iodo-7-methyl-7H-pyrrole[2,3-d]pyrimidine-4-amine.

[0360] 1 H NMR (400MHz, DMSO-d6) δ8.96(t,J=5.9Hz,1H),8.13(s,1H),8.12(d,J=2.0Hz,1H),7.77(dd,J=7.6,1.9Hz,1H),7.33(d,J=7.6Hz,1H),7. 22(dd,J=4.8,1.8Hz,1H),7.13(s,1H),7.07–7.02(m,2H),6.63(s,2H),4.55(d,J=5.7Hz,2H),2.52(s,3H),1.52(s,9H).LR-MS(ESI)m / z 488.2(M+1).

[0361] Example 75 Preparation of 3-((4-amino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0362] Except that compound 74 is used instead of 67-IM, the synthesis method is as described in Example 67.

[0363] 1 H NMR (400MHz, DMSO-d6) δ13.15 (s, 1H), 9.06 (t, J = 6.0Hz, 1H), 8.41 (s, 1H), 8. 14(d,J=1.9Hz,1H),7.94(d,J=2.4Hz,1H),7.82(dd,J=8.0,2.0Hz,1H),7.49 (dd,J=5.0,3.0Hz,1H),7.43(d,J=8.1Hz,1H),7.33(d,J=2.9Hz,1H),7.11–7 .06(m,1H),6.70(s,2H),4.46(d,J=5.8Hz,2H),2.51(s,3H).LR-MS(ESI)m / z 388.1(M+1).

[0364] Example 76 Preparation of 3-((4-amino-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0365] Except for replacing iodomethane with iodomethane in step 1, the synthesis method is as described in Example 72. 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.8Hz,1H),8.19(s,1H),8.05(s,1H),7.84–7.71(m,2H),7.49(dd,J=5.0,2.9Hz,1H),7.42(d,J=8.0Hz,1H),7 .33(s,1H),7.09(d,J=5.4Hz,1H),6.68(s,2H),4.46(d,J=5.8Hz,2H),4.18(q,J=7.2Hz,2H),2.52(s,3H),1.36(t,J=7.2Hz,3H).LR-MS(ESI)m / z 416.1(M+1).

[0366] Example 77 Preparation of 3-((4-amino-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0367] Except for replacing iodomethane with 2-iodopropane in step 1, the synthesis method is as described in Example 72. 1H NMR(400MHz,DMSO-d6)δ9.03(t,J=5.9Hz,1H),8.15(s,1H),8.05(s,1H),7.86(s, 1H),7.78(dd,J=8.0,1.9Hz,1H),7.48(dd,J=5.0,3.0Hz,1H),7.41(d,J=8.0Hz,1 H),7.33(d,J=2.8Hz,1H),7.09(d,J=4.9Hz,1H),6.66(s,2H),4.92(hept,J=7.7H z,1H),4.46(d,J=5.8Hz,2H),2.52(s,3H),1.45(d,J=6.7Hz,6H).LR-MS(ESI)m / z 430.2(M+1).

[0368] Example 78 3-((4-amino-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0369] Except for replacing 5-iodo-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine-4-amine with 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine-4-amine (synthesis method referred to WO2014184069), the synthesis method is as described in Example 72.

[0370] 1 H NMR(400MHz, DMSO-d6)δ8.96(t,J=5.7Hz,1H),8.12(d,J=2.0Hz,1H),8.10( s,1H),7.77(dd,J=7.6,1.9Hz,1H),7.36–7.30(m,1H),7.22(dd,J=4.8,1.8H z,1H),7.09–7.01(m,2H),6.62(s,2H),4.85(hept,J=4.5Hz,1H),4.55(d,J= 5.6Hz,2H),2.51(s,3H),2.27(s,3H),1.32(d,J=4.4Hz,6H).LR-MS(ESI)m / z 444.2(M+1).

[0371] Example 79 Preparation of 3-((4-acetamido-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0372] Compound 72 (80 mg, 0.2 mmol) and AcOH / Ac2O (1 mL / 1 mL) were added to a round-bottom flask and reacted at 80 °C for 8 h. After the reaction was completed, the solvent was evaporated under reduced pressure and separated by column chromatography to obtain 40 mg of 3-((4-acetamido-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide (yield: 45%).

[0373] 1 H NMR (400MHz, DMSO-d6) δ10.22(s,1H),9.02(t,J=6.0Hz,1H),8.65(s,1H),7.99(s,2H),7.78(d,J=9.0Hz,1H),7.49(dd,J=5.4,2.9Hz,1H) ,7.41(d,J=8.1Hz,1H),7.33(s,1H),7.09(d,J=5.4Hz,1H),4.46(d,J=5.7Hz,2H),3.84(s,3H),2.51(s,3H),2.16(s,3H).LR-MS(ESI)m / z 444.1(M+1).

[0374] Example 80 Preparation of 4-methyl-3-((7-methyl-4-(methylamino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-N-(thiophen-3-ylmethyl)benzamide

[0375] The synthesis method is as described in Example 72, except that 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine-4-amine is used instead of 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine-4-amine.

[0376] 1 H NMR (400MHz, DMSO-d6) δ9.01(t,J=5.9Hz,1H),8.15(s,1H),8.05(s,1H),7.86(s,1H),7.78(dd,J=8.0,1.9Hz, 1H),7.48(dd,J=5.0,3.0Hz,1H),7.41(d,J=8.0Hz,1H),7.33(d,J=2.8Hz,1H),7.09(d,J=4.9Hz,1H),5.66(br s,1H),4.46(d,J=5.7Hz,2H),3.95(s,3H),2.73(d,J=2.6Hz,3H),2.52(s,3H).LR-MS(ESI)m / z 416.1(M+1).

[0377] Example 81 Preparation of 4-methyl-3-((7-methyl-4-(isopropylamino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-N-(thiophene-3-ylmethyl)benzamide

[0378] The synthesis method is as described in Example 72, except that 5-iodo-N-isopropyl-7-methyl-7H-pyrrolo[2,3-d]pyrimidine-4-amine is used instead of 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine-4-amine.

[0379] 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=6.0Hz,1H),8.16(s,1H),8.05(s,1H),7.86(s,1H),7.78(dd,J=8.0,1.9Hz, 1H),7.48(dd,J=5.0,3.0Hz,1H),7.41(d,J=8.0Hz,1H),7.33(d,J=2.8Hz,1H),7.09(d,J=4.9Hz,1H),5.46(br s,1H),4.45(d,J=5.9Hz,2H),3.15–2.98(m,1H),2.73(d,J=2.6Hz,3H),2.52(s,3H),1.17(d,J=5.9Hz,6H).LR-MS(ESI)m / z 444.2(M+1).

[0380] Example 82 Preparation of 3-((4-amino-1-methyl-1H-pyrazol[3,4-d]pyrimidin-3-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0381] The synthesis method is as described in Example 72, except that 3-iodo-1-methyl-1H-pyrazole[3,4-d]pyrimidine-4-amine is used instead of 5-iodo-7-methyl-7H-pyrrole[2,3-d]pyrimidine-4-amine.

[0382] 1 H NMR (400MHz, DMSO-d6) δ9.06(t,J=5.7Hz,1H),8.85(s,2H),8.27(s,1H),8.20(s,1H),7.86(d,J=8.0Hz,1H),7.52 –7.41(m,2H),7.34(s,1H),7.09(d,J=5.0Hz,1H),4.47(d,J=5.8Hz,2H),3.94(s,3H),2.53(s,3H).LR-MS(ESI)m / z 403.1(M+1).

[0383] Example 83 Preparation of 3-((4-amino-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0384] Except that the corresponding halide (BrCH2CF3) is used instead of iodomethane in step 1, the synthesis method is as described in Example 72. 1 H NMR(400MHz, DMSO-d6)δ9.03(t,J=5.8Hz,1H),8.23(s,1H),8.09(d,J=1.9Hz,1H ),7.81(dd,J=7.9,2.0Hz,1H),7.76(s,1H),7.49(dd,J=5.0,3.0Hz,1H),7.43(d ,J=8.1Hz,1H),7.33(dd,J=2.9,1.2Hz,1H),7.09(dd,J=4.9,1.3Hz,1H),6.82(s ,2H),5.10(q,J=9.1Hz,2H),4.47(d,J=5.8Hz,2H),2.51(s,3H).LR-MS(ESI)m / z 470.1(M+1).

[0385] Example 84 Preparation of 3-((4-amino-7-(1,1,1-trifluoropropane-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0386] Except that the corresponding halide (CH3CHBrCF3) is used instead of iodomethane in step 1, the synthesis method is as described in Example 72. 1 H NMR (400MHz, DMSO-d6) δ9.02(t,J=5.9Hz,1H),8.23(s,1H),8.11(d,J=1.9Hz,1H),7. 82(dd,J=7.9,2.0Hz,1H),7.76(s,1H),7.49(dd,J=5.0,3.0Hz,1H),7.43(d,J=8.1Hz ,1H),7.33(dd,J=2.8,1.3Hz,1H),7.09(dd,J=4.9,1.3Hz,1H),6.66(s,2H),5.02–4. 89(m,1H),4.47(d,J=5.8Hz,2H),2.52(s,3H),1.39(d,J=6.4Hz,3H).LR-MS(ESI)m / z 484.1(M+1).

[0387] Example 85 Preparation of 3-((4-amino-7-(2-methoxyethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0388] Except that the corresponding halide (CH3OCH2CH2Br) is used instead of iodomethane in step 1, the synthesis method is as described in Example 72. 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.8Hz,1H),8.24(s,1H),8.06(d,J=1.9Hz,1H),7.79( dd,J=8.0,1.9Hz,1H),7.70(s,1H),7.49(dd,J=4.9,2.9Hz,1H),7.42(d,J=8.1Hz,1H),7. 33(dd,J=3.0,1.2Hz,1H),7.09(dd,J=5.0,1.3Hz,1H),6.69(s,2H),4.46(d,J=5.8Hz,2H ),4.31(t,J=5.3Hz,2H),3.69(t,J=5.3Hz,2H),3.24(s,3H),2.51(s,3H).LR-MS(ESI)m / z 446.2(M+1).

[0389] Example 86 Preparation of 3-((4-amino-7-(1-methoxypropane-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0390] Except that the corresponding halide (CH3OCH2CH(CH3)Br) is used instead of iodomethane in step 1, the synthesis method is as described in Example 72. 1 H NMR (400MHz, DMSO-d6) δ8.96(t,J=5.9Hz,1H),8.12(d,J=2.0Hz,1H),8.10(s,1H),7.77(d d,J=7.6,1.9Hz,1H),7.54(s,1H),7.33(dd,J=7.5,1.0Hz,1H),7.22(dd,J=4.8,1.8Hz,1H ),7.08–7.02(m,2H),6.66(s,2H),4.55(d,J=5.7Hz,2H),4.27–4.12(m,1H),4.10–3.95(m ,1H),3.27(s,3H),3.19–3.04(m,1H),2.51(s,3H),1.34(d,J=6.2Hz,3H).LR-MS(ESI)m / z 460.2(M+1).

[0391] Example 87 Preparation of 3-((4-amino-7-(2-(dimethylamino)ethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0392] Except that the corresponding halide ((CH3)2NCH2CH2Br) is used instead of iodomethane in step 1, the synthesis method is as described in Example 72. 1 H NMR(400MHz, DMSO-d6)δ9.04(t,J=5.9Hz,1H),8.21(s,1H),8.06(d,J=2.1H z,1H),7.85–7.72(m,2H),7.49(dd,J=4.9,2.9Hz,1H),7.43(d,J=8.0Hz,1H) ,7.33(s,1H),7.09(d,J=5.0Hz,1H),6.78(s,2H),4.56–4.48(m,2H),4.47(d ,J=5.8Hz,2H),3.61–3.42(m,2H),2.77(s,6H),2.51(s,3H).LR-MS(ESI)m / z 459.2(M+1).

[0393] Example 88 Preparation of 3-((4-amino-7-(1-(dimethylamino)propane-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0394] Except that the corresponding halide ((CH3)2NCH2CH(CH3)Br) is used instead of iodomethane in step 1, the synthesis method is as described in Example 72. 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.9Hz,1H),8.15(s,1H),8.06(d,J=1.9Hz,1H),7.79(d d,J=8.0,1.9Hz,1H),7.70(s,1H),7.48(dd,J=5.0,3.0Hz,1H),7.42(d,J=8.0Hz,1H),7.3 6–7.30(m,1H),7.09(dd,J=5.0,1.3Hz,1H),6.66(s,2H),4.46(d,J=5.8Hz,2H),4.27–4.1 2(m,1H),4.10–3.95(m,1H),3.19–3.04(m,1H),2.18(s,6H),0.87(s,3H).LR-MS(ESI)m / z 473.2(M+1).

[0395] Example 89 Preparation of 3-((4-amino-7-cyclobutyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0396] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.9Hz,1H),8.15(s,1H),8.06(d,J=1.9Hz,1H),8.00(s,1H) ,7.79(dd,J=8.0,1.9Hz,1H),7.49(dd,J=5.0,3.0Hz,1H),7.42(d,J=8.1Hz,1H),7.33(dd,J=3 .0,1.3Hz,1H),7.09(dd,J=4.9,1.3Hz,1H),6.68(s,2H),5.15(p,J=8.6Hz,1H),4.47(d,J=5.8 Hz,2H),2.61–2.52(m,2H),2.51(s,3H),2.44–2.35(m,2H),1.88–1.78(m,2H).LR-MS(ESI)m / z 442.2(M+1).

[0397] Example 90 Preparation of tert-butyl-3-(4-amino-5-((2-methyl-5-((thiophen-3-ylmethyl)carbamoyl)phenyl)ethynyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)azacyclobutane-1-carbonate

[0398] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR (400MHz, chloroform-d) δ8.29(s,1H),7.90(d,J=2.0Hz,1H),7.66(dd,J=8.0,2.0Hz,1H),7.55( s,1H),7.36–7.29(m,2H),7.24(dd,J=3.0,1.2Hz,4H),7.10(dd,J=4.9,1.3Hz,1H),6.39(t ,J=5.5Hz,1H),5.70(s,2H),5.60–5.49(m,1H),4.66(d,J=5.5Hz,2H),4.49(dd,J=9.6,8.1 Hz, 2H), 4.20 (dd, J=9.6, 5.0Hz, 2H), 2.55 (s, 3H), 1.48 (s, 9H). LR-MS (ESI) m / z543.2 (M+1).

[0399] Example 91 Preparation of 3-((4-amino-7-(1-ethylazacyclobutane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0400]

[0401] Step 1: Add compound 90 (0.3 g, 0.55 mmol) and DCM (5 mL) to a round-bottom flask, then add HCl·dioxane solution (0.5 mL, 2 mmol) and stir overnight at room temperature. After the reaction is complete, evaporate the solvent under reduced pressure to obtain 0.26 g of yellow solid 91-IM (yield: 98.2%), which can be used directly in the next step without further purification.

[0402] Step 2: Add 106-IM (80 mg, 0.17 mmol), K2CO3 (115 mg, 0.84 mmol), and anhydrous DMF (2 mL) obtained in the previous step to a round-bottom flask, and add IEt (16 μL, 0.2 mmol) dropwise. React overnight at room temperature. After the reaction is complete, pour the reaction solution into water, extract with ethyl acetate, combine the organic phases, wash with H2O and NaCl aqueous solution respectively, dry with anhydrous Na2SO4, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 25 mg of 3-((4-amino-7-(1-ethylazacyclobutane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide (white solid; yield: 32%). 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.9Hz,1H),8.16(s,1H),8.07(d,J=1.9Hz,1H),8.03(s,1H),7.8 0(dd,J=8.0,2.0Hz,1H),7.49(dd,J=4.9,2.9Hz,1H),7.42(d,J=8.0Hz,1H),7.33(dd,J=2.9,1.2Hz ,1H),7.09(dd,J=5.0,1.3Hz,1H),6.73(s,2H),5.29–5.18(m,1H),4.47(d,J=5.8Hz,2H),3.83–3.6 7(m,2H),3.56–3.37(m,2H),2.66–2.54(m,2H),2.52(s,3H),0.95(t,J=7.1Hz,3H).LR-MS(ESI)m / z 471.2(M+1).

[0403] Example 92 Preparation of 3-((4-amino-7-(1-isopropylazacyclobutane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0404] The synthesis method is as described in Example 91. 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=6.0Hz,1H),8.17(s,1H),8.05(d,J=2.0Hz,1H),8.03(s,1H),7.81 (d,J=8.0Hz,1H),7.51(dd,J=4.9,2.9Hz,1H),7.42(d,J=8.0Hz,1H),7.33(dd,J=2.9,1.2Hz,1H),7.0 9(dd,J=5.0,1.3Hz,1H),6.67(s,2H),5.27–5.18(m,1H),4.45(d,J=5.9Hz,2H),3.83–3.67(m,2H),3 .52–3.34(m,2H),2.88–2.65(m,1H),2.52(s,3H),1.05(d,J=5.1Hz,6H).LR-MS(ESI)m / z485.2(M+1).

[0405] Example 93 Preparation of 3-((4-amino-7-(1-cyclopropylazacyclobutane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0406] The synthesis method is as described in Example 91. 1 H NMR (400MHz, DMSO-d6) δ9.01(t,J=6.0Hz,1H),8.14(s,1H),8.09(s,1H),8.03(s,1H),7.80(dd,J=8. 0,2.0Hz,1H),7.51(dd,J=5.1,2.5Hz,1H),7.42(d,J=8.0Hz,1H),7.33(d,J=2.9Hz,1H),7.11(d,J=5. 0Hz,1H),6.68(s,2H),5.27–5.16(m,1H),4.45(d,J=5.9Hz,2H),3.83–3.67(m,2H),3.56–3.37(m,2H ),2.52(s,3H),2.46–2.34(m,1H),0.90–0.76(m,2H),0.64–0.52(m,2H).LR-MS(ESI)m / z483.2(M+1).

[0407] Example 94 Preparation of 3-((4-amino-7-(1-acetylazonobutane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0408] Compound 91-IM (80 mg, 0.17 mmol), Et3N (70 μL, 0.5 mmol), and anhydrous DCM (3 mL) were added to a round-bottom flask. Acetyl chloride (13 μL, 0.18 mmol) was added dropwise in an ice bath, and the reaction was carried out at room temperature for 1 h. After quenching the reaction with 0.2 mL of methanol, the solvent was evaporated under reduced pressure, and the product was separated by column chromatography to obtain 15 mg of 3-((4-amino-7-(1-acetylazetane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide (pale yellow solid; yield: 18.5%). 1 H NMR(400MHz, DMSO-d6)δ9.04(t,J=5.7Hz,1H),8.17(s,1H),8.06(s,1H),7.79(d, J=8.0Hz,1H),7.70(s,1H),7.49(s,1H),7.42(d,J=8.0Hz,1H),7.34(s,1H),7.09( d,J=5.0Hz,1H),6.71(s,2H),4.96–4.80(m,1H),4.47(d,J=5.8Hz,2H),4.12–3.96 (m,2H),3.93–3.76(m,2H),2.52(s,3H),2.01(s,3H).LR-MS(ESI)m / z484.2(M+1).

[0409] Example 95 Preparation of 3-((4-amino-7-(1-methanesulfonylazonobutane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0410] Except for replacing acetyl chloride with methanesulfonyl chloride, the synthesis method is as described in Example 94. 1H NMR(400MHz,DMSO-d6)δ9.03(t,J=5.9Hz,1H),8.15(s,1H),8.05(s,1H),7.86(s,1H), 7.78(dd,J=8.0,1.9Hz,1H),7.48(dd,J=5.0,3.0Hz,1H),7.41(d,J=8.0Hz,1H),7.33(d ,J=2.8Hz,1H),7.09(d,J=4.9Hz,1H),6.66(s,2H),4.92–4.79(m,1H),4.46(d,J=5.8H z,2H),4.15–3.99(m,2H),3.94–3.78(m,2H),2.89(s,3H),2.52(s,3H).LR-MS(ESI)m / z 521.1(M+1).

[0411] Example 96 Preparation of 3-((4-amino-7-(3-oxocyclobutyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0412] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR (400MHz, chloroform-d) δ8.31(s,1H),7.91(d,J=2.0Hz,1H),7.65(dd,J=8.0,2.0Hz,1H),7.37–7.30(m,3H),7.24(dd,J=2.9,1.2Hz,1H),7.10(dd,J =4.9,1.3Hz,1H),6.35(t,J=5.7Hz,1H),5.66(s,2H),5.39–5.32(m,1H),4.66(d,J=5.5Hz,2H),3.87–3.63(m,4H),2.54(s,3H).LR-MS(ESI)m / z 456.1(M+1).

[0413] Example 97 Preparation of 3-((4-amino-7-(3-hydroxycyclobutyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0414] Compound 96 (50 mg, 0.11 mmol) and anhydrous THF (2 mL) were added to a round-bottom flask, followed by NaBH4 (12 mg, 0.33 mmol). The mixture was reacted at room temperature for 2 h. An aqueous solution of NH4Cl was added, and the solvent was evaporated under reduced pressure. The product was then separated by column chromatography to obtain 20 mg of 3-((4-amino-7-(3-hydroxycyclobutyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide (white solid; yield: 40%). 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.9Hz,1H),8.14(s,1H),8.06(d,J=1.9Hz,1H),7.93(s,1 H),7.79(d,J=8.1Hz,1H),7.49(dd,J=4.9,2.9Hz,1H),7.42(d,J=8.1Hz,1H),7.33(s,1H),7 .09(d,J=4.9Hz,1H),6.69(s,2H),5.28(d,J=7.0Hz,1H),4.73–4.63(m,1H),4.47(d,J=5.8H z,2H),4.08–3.92(m,1H),2.84–2.73(m,2H),2.52(s,3H),2.38–2.27(m,2H).LR-MS(ESI)m / z 458.2(M+1).

[0415] Example 98 Preparation of 3-((4-amino-7-(oxecyclobutane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0416] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz, DMSO-d6)δ8.96(t,J=6.0Hz,1H),8.12(d,J=2.0Hz,1H),8.10(s,1H),7.77(dd, J=7.6,1.9Hz,1H),7.54(s,1H),7.33(dd,J=7.5,1.0Hz,1H),7.22(dd,J=4.8,1.8Hz,1H),7. 06(t,J=1.6Hz,1H),7.04(dd,J=4.9,1.6Hz,1H),6.69(s,2H),5.13–5.04(m,2H),4.96–4.88 (m,1H),4.88–4.81(m,2H),4.55(d,J=5.9Hz,2H),2.52(s,3H).LR-MS(ESI)m / z444.1(M+1).

[0417] Example 99 Preparation of 3-((4-amino-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0418] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR (400MHz, DMSO-d6) δ9.04(t,J=5.9Hz,1H),8.17(s,1H),8.05(d,J=1.9Hz,1H),7.83(s ,1H),7.79(dd,J=8.0,1.9Hz,1H),7.49(dd,J=4.9,2.9Hz,1H),7.42(d,J=8.0Hz,1H),7.3 6–7.31(m,1H),7.09(dd,J=5.0,1.3Hz,1H),6.68(s,2H),5.04(m,1H),4.47(d,J=5.8Hz,2 H),2.52(s,3H),2.11(m,2H),1.95–1.82(m,4H),1.69(m,2H).LR-MS(ESI)m / z456.2(M+1).

[0419] Example 100 Preparation of 3-((4-amino-7-(tetrahydrofuran-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0420] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz, DMSO-d6)δ8.96(t,J=6.0Hz,1H),8.12(d,J=2.0Hz,1H),8.10(s,1H),7. 77(dd,J=7.6,1.9Hz,1H),7.54(s,1H),7.33(dd,J=7.5,1.0Hz,1H),7.22(dd,J=4.8,1 .8Hz,1H),7.07–7.01(m,2H),6.68(s,2H),4.55(d,J=5.9Hz,2H),4.18–4.00(m,2H), 3.98–3.76(m,3H),2.52(s,3H),2.26–2.14(m,1H),2.02–1.88(m,1H).LR-MS(ESI)m / z 458.2(M+1).

[0421] Example 101 Preparation of tert-butyl-4-(4-amino-5-((2-methyl-5-((thiophen-3-yl)carbamoyl)phenyl)ethynyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carbonate

[0422] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 ¹H NMR (400MHz, chloroform-d) δ 8.29 (s, 1H), 7.90 (d, J = 2.0 Hz, 1H), 7.65 (dd, J = 8.0, 2.0 Hz, 1H), 7.32–7.27 (m, 2H), 7.27 (s, 1H), 7.22–7.20 (m, 1H), 7.08 (dd, J = 5.0, 1.3 Hz, 1H), 6.61 (t, J = 5.6 Hz, 1H) ),5.79(s,2H),4.79(tt,J=12.1,4.0Hz,1H),4.64(d,J=5.6Hz,2H),4.41–4.19(m,2H),2.98– 2.81(m,2H),2.52(s,3H),2.08–2.01(m,2H),1.89–1.80(m,2H),1.48(s,9H).LR-MS(ESI)m / z 571.2(M+1).

[0423] Example 102 Preparation of 3-((4-amino-7-(1-methylpiperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0424] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.9Hz,1H),8.15(s,1H),8.05(d,J=1.9Hz,1H),7.85(s,1H) ,7.79(dd,J=8.0,1.9Hz,1H),7.48(dd,J=5.0,3.0Hz,1H),7.42(d,J=8.0Hz,1H),7.33(d,J=2. 9Hz,1H),7.09(d,J=4.9Hz,1H),6.67(s,2H),4.56–4.48(m,1H),4.46(d,J=5.8Hz,2H),2.90(d ,J=7.1Hz,2H),2.51(s,3H),2.22(s,3H),2.12–1.96(m,4H),1.91–1.80(m,2H).LR-MS(ESI)m / z 485.2(M+1).

[0425] Example 103 Preparation of 3-((4-amino-7-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0426] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz, DMSO-d6)δ8.96(t,J=6.0Hz,1H),8.10(s,1H),8.02(d,J=2.0Hz,1H), 7.77(dd,J=7.6,1.9Hz,1H),7.54(s,1H),7.33(dd,J=7.5,1.0Hz,1H),7.26–7.18(m ,1H),7.09–7.01(m,2H),6.64(s,2H),4.48(d,J=5.9Hz,2H),3.90–3.73(m,3H),3. 52–3.34(m,2H),2.53(s,3H),2.04–1.88(m,2H),1.79–1.64(m,2H).LR-MS(ESI)m / z 472.2(M+1).

[0427] Example 104 Preparation of 3-((4-amino-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0428] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz,DMSO-d6)δ9.03(t,J=5.9Hz,1H),8.15(s,1H),8.05(s,1H),7.86(s,1 H),7.78(dd,J=8.0,1.9Hz,1H),7.48(dd,J=5.0,3.0Hz,1H),7.41(d,J=8.0Hz,1H), 7.33(d,J=2.8Hz,1H),7.09(d,J=4.9Hz,1H),6.62(s,2H),5.01-4.92(m,1H),4.46( d,J=5.8Hz,2H),2.52(s,3H),0.64–0.51(m,2H),0.41–0.26(m,2H).LR-MS(ESI)m / z 428.1(M+1).

[0429] Example 105 Preparation of 3-((4-amino-7-(pentan-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0430] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.9Hz,1H),8.15(s,1H),8.05(d,J=1.9Hz,1H ),7.85(s,1H),7.79(dd,J=8.0,1.9Hz,1H),7.48(dd,J=5.0,3.0Hz,1H),7.42(d, J=8.0Hz,1H),7.33(d,J=2.9Hz,1H),7.09(d,J=4.9Hz,1H),6.67(s,2H),4.56–4. 47(m,1H),2.52(s,3H),1.47–1.36(m,4H),0.83(t,J=4.5Hz,6H).LR-MS(ESI)m / z 458.2(M+1).

[0431] Example 106 Preparation of 3-((4-amino-7-isobutyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide

[0432] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz,DMSO-d6)δ9.02(t,J=5.8Hz,1H),8.19(s,1H),8.05(s,1H),7.8 4–7.71(m,2H),7.49(dd,J=5.0,2.9Hz,1H),7.42(d,J=8.0Hz,1H),7.33(s,1H ),7.09(d,J=5.4Hz,1H),6.68(s,2H),4.44(d,J=5.8Hz,2H),4.02(d,J=6.9H z,2H),2.51(s,3H),2.05–1.89(m,1H),0.91(d,J=4.6Hz,6H).LR-MS(ESI)m / z 444.2(M+1).

[0433] Example 107 Preparation of 3-((4-amino-7-((3-methyloxetane-3-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0434] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz,DMSO-d6)δ9.03(t,J=5.9Hz,1H),8.12(s,1H),8.05(s,1H),7.86(s,1H),7.7 7(dd,J=8.0,1.9Hz,1H),7.48(dd,J=5.0,3.0Hz,1H),7.41(d,J=8.0Hz,1H),7.33(d,J=2.8 Hz,1H),7.09(d,J=4.9Hz,1H),6.64(s,2H),4.46(d,J=5.8Hz,2H),4.29(d,J=9.3Hz,2H), 4.14(d,J=9.4Hz,2H),3.91(s,2H),2.52(s,3H),0.97(s,3H).LR-MS(ESI)m / z472.2(M+1).

[0435] Example 108 Preparation of 3-((7-(2-acetamidoethyl)-4-amino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0436] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz, DMSO-d6)δ9.04(t,J=5.9Hz,1H),8.21(s,1H),8.06(d,J=2.1Hz,1H), 7.97(t,J=5.6Hz,1H),7.85–7.72(m,2H),7.49(dd,J=4.9,2.9Hz,1H),7.43(d,J=8 .0Hz,1H),7.33(s,1H),7.09(d,J=5.0Hz,1H),6.78(s,2H),4.69(t,J=6.5Hz,2H), 4.45(d,J=5.8Hz,2H),3.35–3.23(m,2H),2.51(s,3H),1.89(s,3H).LR-MS(ESI)m / z 473.2(M+1).

[0437] Example 109 Preparation of 3-((4-amino-7-(3-dimethylamino)-3-oxopropyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0438] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72.1 H NMR(400MHz, DMSO-d6)δ9.01(t,J=6.0Hz,1H),8.17(s,1H),8.06(d,J=2.1Hz, 1H),7.85–7.72(m,2H),7.49(dd,J=4.9,2.9Hz,1H),7.43(d,J=8.0Hz,1H),7.3 1(s,1H),7.09(d,J=5.0Hz,1H),6.68(s,2H),4.83(t,J=10.2Hz,2H),4.47(d,J =6.0Hz,2H),2.76(s,6H),2.52(s,6H),2.41(t,J=10.0Hz,2H).LR-MS(ESI)m / z 487.2(M+1).

[0439] Example 110 Preparation of 3-((4-amino-7-(2-fluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0440] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz,DMSO-d6)δ9.03(t,J=5.8Hz,1H),8.24(s,1H),8.06(d,J=1.9Hz,1H),7 .79(dd,J=8.0,1.9Hz,1H),7.70(s,1H),7.49(dd,J=4.9,2.9Hz,1H),7.42(d,J=8.1 Hz,1H),7.33(dd,J=3.0,1.2Hz,1H),7.09(dd,J=5.0,1.3Hz,1H),6.69(s,2H),4.46 –4.26(m,2H),4.31(t,J=5.5Hz,2H),4.12–3.98(m,2H),2.51(s,3H).LR-MS(ESI)m / z 434.1(M+1).

[0441] Example 111 Preparation of 3-((4-amino-7-hydroxyethyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0442] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.9Hz,1H),8.21(s,1H),8.06(d,J=1.9Hz,1H),7.77(d d,J=8.0,2.0Hz,1H),7.70(s,1H),7.49(dd,J=4.9,2.9Hz,1H),7.42(d,J=8.1Hz,1H),7.33 (dd,J=3.0,1.2Hz,1H),7.09(dd,J=5.0,1.3Hz,1H),6.69(s,2H),4.96(t,J=6.5Hz,1H),4. 46(d,J=5.8Hz,2H),4.32(t,J=7.1Hz,2H),3.52–3.43(m,2H),2.53(s,3H).LR-MS(ESI)m / z 432.1(M+1).

[0443] Example 112 Preparation of 3-((4-amino-7-(2-hydroxy-2-methylpropyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0444] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz, DMSO-d6)δ9.03(t,J=5.9Hz,1H),8.15(s,1H),8.06(d,J=1.9Hz,1H),8 .00(s,1H),7.79(dd,J=8.0,1.9Hz,1H),7.49(dd,J=5.0,3.0Hz,1H),7.42(d,J=8.1H z,1H),7.33(dd,J=3.0,1.3Hz,1H),7.09(dd,J=4.9,1.3Hz,1H),6.68(s,2H),4.67( s,1H),4.47(d,J=5.8Hz,2H),3.92(s,2H),2.51(s,3H),1.23(s,6H).LR-MS(ESI)m / z 460.2(M+1).

[0445] Example 113 Preparation of 3-((4-amino-7-((2R,5S)-5-(hydroxymethyl)tetrahydrofuran-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0446] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1H NMR(400MHz, DMSO-d6)δ8.96(t,J=6.0Hz,1H),8.12(d,J=2.0Hz,1H),8.10(s,1H),7.77(dd,J=7.6,1.9Hz,1H),7 .54(s,1H),7.33(dd,J=7.5,1.0Hz,1H),7.22(dd,J=4.8,1.8Hz,1H),7.07–7.01(m,2H),6.62(s,2H),4.92–4.83 (m,1H),4.55(d,J=5.9Hz,2H),4.50(t,J=7.4Hz,1H),4.42–4.31(m,1H),3.79(ddd,J=11.8,7.4,6.3Hz,1H),3.5 4(ddd,J=11.9,7.3,6.2Hz,1H),2.52(s,3H),2.22–2.06(m,2H),1.98–1.85(m,2H).LR-MS(ESI)m / z488.2(M+1).

[0447] Example 114 Preparation of 3-((4-amino-7-(3,3-difluorocyclobutyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0448] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz,DMSO-d6)δ9.03(t,J=5.9Hz,1H),8.14(s,1H),8.06(d,J=1.9Hz,1H) ,7.93(s,1H),7.79(d,J=8.1Hz,1H),7.49(dd,J=4.9,2.9Hz,1H),7.42(d,J=8.1Hz ,1H),7.33(s,1H),7.09(d,J=4.9Hz,1H),6.69(s,2H),4.73–4.63(m,1H),4.47(d ,J=5.8Hz,2H),2.73–2.54(m,2H),2.51(s,3H),2.47–2.25(m,2H).LR-MS(ESI)m / z 478.1(M+1).

[0449] Example 115 Preparation of 3-((4-amino-7-(3,6-dihydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0450] The synthesis method is as described in Example 72, except that 7-(3,6-dihydro-2H-pyran-4-yl)-5-iodo-7H-pyrrolo[2,3-d]pyrimidine-4-amine is used instead of 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidine-4-amine. 1 H NMR (400MHz, DMSO-d6) δ9.03(t,J=5.9Hz,1H),8.14(s,1H),8.06(d,J=1.9Hz,1H),7.93 (s,1H),7.79(d,J=8.1Hz,1H),7.49(dd,J=4.9,2.9Hz,1H),7.42(d,J=8.1Hz,1H),7.33 (s,1H),7.09(d,J=4.9Hz,1H),6.69(s,2H),5.67–5.52(m,1H),4.47(d,J=5.8Hz,2H),4 .17(d,J=4.6Hz,2H),3.92–3.71(m,2H),2.51(s,3H),2.01–1.84(m,2H).LR-MS(ESI)m / z 470.2(M+1).

[0451] The synthetic route for the intermediate 7-(3,6-dihydro-2H-pyran-4-yl)-5-iodo-7H-pyrrole[2,3-d]pyrimidine-4-amine is shown below.

[0452]

[0453] Step 1: Add 7H-pyrrolo[2,3-d]pyrimidine-4-amine (1.0 g, 7.45 mmol), DMF-DMA (1.07 g, 8.95 mmol), and DMF (20 mL) to a round-bottom flask and react overnight at room temperature. Evaporate the solvent under reduced pressure to obtain the crude product, redissolve it in DCM, wash with H2O and NaCl aqueous solution respectively, dry with anhydrous Na2SO4, and separate by column chromatography to obtain compound 115-2 950 mg (yield: 67.4%). 1 H NMR (400MHz, DMSO-d6) δ12.06(s,1H),8.73(s,1H),8.20(s,1H),7.19(d,J=2.7Hz,1H ),6.62(dd,J=3.4,1.9Hz,1H),2.87(s,3H),2.83(s,3H).LR-MS(ESI)m / z190.1(M+1).

[0454] Step 2: Add compound 115-2 (250 mg, 1.32 mmol), pinacol 3,6-dihydro-2H-pyran-4-boronate (333 mg, 1.59 mmol), copper acetate (291 mg, 1.59 mmol), 2,2'-bipyridine (248 mg, 1.59 mmol), anhydrous Na₂CO₃ (420 mg, 3.96 mmol), and DMAC (10 mL) to a round-bottom flask. Stir at 90 °C for 4 h, add DCM, and wash three times with water. Dry the organic phase with anhydrous Na₂SO₄ and evaporate under reduced pressure to obtain intermediate 115-3, which requires no further purification.

[0455] Step 3: Add EtOH (10 mL) and ethylenediamine (0.2 mL) to the intermediate 115-3 from the previous step, and reflux for 16 h. After the reaction is complete, evaporate the solvent under reduced pressure, redissolve the compound in DCM, wash with NaCl aqueous solution, dry with anhydrous Na2SO4, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain 160 mg of compound 115-4 (overall yield of two steps: 56%). 1 H NMR (400MHz, DMSO-d6) δ8.71(s,1H),7.70(d,J=5.7Hz,1H),7.52(d,J=6.0Hz,1H),6.72(s,2H ),5.20–4.97(m,1H),4.20–4.10(m,2H),3.88–3.82(m,2H),2.17–2.05(m,2H).LR-MS(ESI)m / z 217.1(M+1).

[0456] Step 4: Compound 115-4 (160 mg, 0.74 mmol), NIS (183 mg, 0.81 mmol), and DMF (3 mL) were added to a round-bottom flask and reacted at room temperature for 4 h. After the reaction was complete, the solvent was evaporated under reduced pressure, and the product was separated by column chromatography to obtain 180 mg of 7-(3,6-dihydro-2H-pyran-4-yl)-5-iodo-7H-pyrrole[2,3-d]pyrimidine-4-amine (yield: 71%). 1 H NMR(400MHz, DMSO-d6)δ8.73(s,1H),7.68(s,1H),6.68(s,2H),5.13–4.96(m,1H),4.21–3.88(m,4H),2.13–2.01(m,2H).LR-MS(ESI)m / z 343.1(M+1).

[0457] Example 116 Preparation of 3-((4-amino-7-((1-methyl-1H-pyrazol-4-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0458] Except that the corresponding halide is used instead of iodomethane in step 1, the synthesis method is as in Example 72. 1 H NMR(400MHz,DMSO-d6)δ9.02(t,J=5.9Hz,1H),8.15(s,1H),8.05(s,1H),7.86( s,1H),7.78(dd,J=8.0,1.9Hz,1H),7.48(dd,J=5.0,3.0Hz,1H),7.41(d,J=8.0 Hz,1H),7.36(s,2H),7.33(d,J=2.8Hz,1H),7.09(d,J=4.9Hz,1H),6.64(s,2H) ,5.57(s,2H),4.46(d,J=6.0Hz,2H),3.94(s,3H),2.52(s,3H).LR-MS(ESI)m / z 482.2(M+1).

[0459] Example 117 Preparation of 3-((4-amino-7-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0460] Except that in step 2, the corresponding diazole derivative is used instead of pinacol 3,6-dihydro-2H-pyran-4-boronic acid ester, the synthesis method is as described in Example 115. 1 H NMR(400MHz, DMSO-d6)δ9.02(t,J=6.0Hz,1H),8.14(s,1H),8.05(d,J=1.9Hz,1H) ,7.93(s,1H),7.79(d,J=8.1Hz,1H),7.52(d,J=1.8Hz,1H),7.49(dd,J=4.9,2.9Hz ,1H),7.42(d,J=8.1Hz,1H),7.33(s,1H),7.30(d,J=1.6Hz,1H),7.09(d,J=4.9Hz, 1H),6.69(s,2H),4.47(d,J=5.8Hz,2H),2.51(s,3H),3.89(s,3H).LR-MS(ESI)m / z 468.2(M+1).

[0461] Example 118 Preparation of 3-((4-amino-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(1-(thiophen-3-yl)cyclopropyl)benzamide

[0462] Except that 2-iodopropane is used instead of iodomethane in step 1, and the corresponding intermediate is obtained in vitro with reference to the method in step 3 of Example 1, the synthesis method is as in Example 72. 1 H NMR (400MHz, DMSO-d6) δ8.86 (s, 1H), 8.12 (d, J = 2.0Hz, 1H), 8.10 (s, 1H), 7.77 (dd ,J=7.6,1.9Hz,1H),7.54(s,1H),7.33(d,J=7.6,1H),7.25(dd,J=5.2,1.9Hz,1H), 7.10–7.05(m,2H),6.64(s,2H),4.94–4.75(m,1H),2.45(d,J=0.9Hz,3H),1.32(d ,J=6.5Hz,6H),0.95–0.79(m,2H),0.71–0.54(m,2H).LR-MS(ESI)m / z456.2(M+1).

[0463] Example 119 Preparation of 3-((4-amino-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl-d2)benzamide

[0464] Except that 2-iodopropane is used instead of iodomethane in step 1, and the corresponding intermediate is obtained in vitro with reference to the method in step 3 of Example 1, the synthesis method is as in Example 72. 1 H NMR(400MHz,DMSO-d6)δ9.03(s,1H),8.17(s,1H),8.05(s,1H),7.86(s,1H), 7.78(dd,J=8.0,1.9Hz,1H),7.48(dd,J=5.0,3.0Hz,1H),7.41(d,J=8.0Hz,1 H),7.33(d,J=2.8Hz,1H),7.09(d,J=4.9Hz,1H),6.65(s,2H),4.92(hept,J= 7.7Hz, 1H), 2.52 (s, 3H), 1.45 (d, J=6.7Hz, 6H). LR-MS (ESI) m / z432.2 (M+1).

[0465] Example 120 Preparation of 3-((4-amino-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophen-2-ylmethyl)benzamide

[0466] Except that 2-iodopropane is used instead of iodomethane in step 1, and the corresponding intermediate is obtained in vitro with reference to the method in step 3 of Example 1, the synthesis method is as in Example 72. 1H NMR(400MHz,DMSO-d6)δ8.98(t,J=6.0Hz,1H),8.12(d,J=2.0Hz,1H),8.10(s,1H),7.7 7(dd,J=7.6,1.9Hz,1H),7.54(s,1H),7.44(dd,J=5.1,1.8Hz,1H),7.33(dd,J=7.5,1. 0Hz,1H),7.12(dd,J=6.4,1.8Hz,1H),7.02(dd,J=6.4,5.1Hz,1H),6.64(s,2H),4.89– 4.81(m,1H),4.45(d,J=5.9Hz,2H),2.53(s,3H),1.34(d,J=4.6Hz,6H).LR-MS(ESI)m / z 430.2(M+1).

[0467] Example 121 Preparation of 3-((4-amino-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-N-((5-chlorothiophene-2-yl)methyl)-4-methylbenzamide

[0468] Except that 2-iodopropane is used instead of iodomethane in step 1, and the corresponding intermediate is obtained in vitro with reference to the method in step 3 of Example 1, the synthesis method is as in Example 72. 1 H NMR (400MHz, DMSO-d6) δ9.04(t,J=8.0Hz,1H),8.36–8.29(m,2H),7.98(dd,J=7.6,1.9Hz,1H),7.75(s,1H),7.54(d,J=7.5Hz,1H),7.33(d,J= 6.4Hz,1H),7.27(d,J=6.4Hz,1H),6.67(s,2H),5.12–5.03(m,1H),4.56(d,J=8.0Hz,2H),2.54(s,3H),1.43(d,J=4.6Hz,6H).LR-MS(ESI)m / z 464.1(M+1).

[0469] Example 122 Preparation of 3-((4-amino-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-N-((5-cyclopropylthiophen-2-yl)methyl)-4-methylbenzamide

[0470] Except that 2-iodopropane is used instead of iodomethane in step 1, and the corresponding intermediate is obtained in vitro with reference to the method in step 3 of Example 1, the synthesis method is as in Example 72. 1H NMR(400MHz,DMSO-d6)δ9.02(t,J=5.8Hz,1H),8.31(d,J=2.0Hz,1H),8.29(s,1 H),7.96(dd,J=7.6,1.9Hz,1H),7.73(s,1H),7.52(dd,J=7.5,1.0Hz,1H),7.15 –7.07(m,2H),6.62(s,2H),5.10–4.98(m,1H),4.54(d,J=6.0Hz,2H),2.64(s,3 H),2.05–1.93(m,1H),1.51(d,J=4.6Hz,6H),0.91–0.70(m,4H).LR-MS(ESI)m / z 470.2(M+1).

[0471] Example 123 Preparation of 3-((4-amino-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-N-((6-fluoropyridin-3-yl)methyl)-4-methylbenzamide

[0472] Except that 2-iodopropane is used instead of iodomethane in step 1, and the corresponding intermediate is obtained in vitro with reference to the method in step 3 of Example 1, the synthesis method is as in Example 72. 1 H NMR(400MHz, DMSO-d6)δ9.03(t,J=5.8Hz,1H),8.42–8.37(m,2H),8.19(d,J=2.0Hz,1H),8.17(s,1H),7.84(dd,J=7.6,1.9Hz,1H),7.61(s,1H),7.40 (dd,J=7.5,1.0Hz,1H),7.23–7.08(m,1H),6.72(s,2H),4.97–4.85(m,1H) ,4.47(d,J=5.7Hz,2H),2.52(s,3H),1.39(d,J=4.7Hz,6H).LR-MS(ESI)m / z 443.2(M+1).

[0473] Example 124 Preparation of 3-((4-amino-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-((6-methylpyridin-3-yl)methyl)benzamide

[0474] Except that 2-iodopropane is used instead of iodomethane in step 1, and the corresponding intermediate is obtained in vitro with reference to the method in step 3 of Example 1, the synthesis method is as in Example 72. 1H NMR (400MHz, DMSO-d6) δ8.96(t,J=6.0Hz,1H),8.55(d,J=2.3Hz,1H),8.12(d,J=2.0Hz ,1H),8.10(s,1H),7.92(dd,J=7.5,2.2Hz,1H),7.77(dd,J=7.6,1.9Hz,1H),7.54(s,1H ),7.33(dd,J=7.5,1.1Hz,1H),7.29(d,J=7.5Hz,1H),6.64(s,2H),4.89–4.82(m,1H), 4.50(d,J=5.7Hz,2H),2.58(s,3H),2.52(s,3H),1.34(d,J=4.9Hz,6H).LR-MS(ESI)m / z 439.2(M+1).

[0475] Example 125 Preparation of 3-((4-amino-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-N-((6-cyclopropylpyridin-3-yl)methyl)-4-methylbenzamide

[0476] Except that 2-iodopropane is used instead of iodomethane in step 1, and the corresponding intermediate is obtained in vitro with reference to the method in step 3 of Example 1, the synthesis method is as in Example 72. 1 H NMR (400MHz, DMSO-d6) δ9.02(t,J=6.0Hz,1H),8.52(d,J=2.1Hz,1H),8.09(d,J=2.0Hz,1H) ,8.04(s,1H),7.92(dd,J=7.5,1.9Hz,1H),7.77(dd,J=7.6,1.9Hz,1H),7.54(s,1H),7.33(d ,J=7.5Hz,1H),7.29(d,J=7.5Hz,1H),6.64(s,2H),4.89–4.82(m,1H),4.45(d,J=5.7Hz,2H ),3.31–2.89(m,1H),2.53(s,3H),1.34(d,J=4.9Hz,6H),0.98–0.73(m,4H).LR-MS(ESI)m / z 465.2(M+1).

[0477] Example 126 Preparation of ethyl-4-amino-5-((2-methyl-5-((thiophen-3-ylmethyl)carbamoyl)phenyl)ethynyl)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylic acid ester

[0478] The synthesis method is as described in Example 72, except that 5-iodo-7-methyl-7H-pyrrole[2,3-d]pyrimidine-7-carbonate (126-3) is used instead of 5-iodo-7-methyl-7H-pyrrole[2,3-d]pyrimidine-4-amine. 1 H NMR(400MHz, DMSO-d6)δ8.96(t,J=5.8Hz,1H),8.13(s,1H),8.12(d,J=2.0Hz,1H ),7.77(dd,J=7.6,1.9Hz,1H),7.72(s,1H),7.33(dd,J=7.5,1.0Hz,1H),7.22(d d,J=4.8,1.8Hz,1H),7.07–7.02(m,2H),6.70(s,2H),4.55(d,J=5.9Hz,2H),4.3 6(q,J=6.2Hz,2H),2.45(d,J=0.9Hz,3H),1.26(t,J=5.9Hz,3H).LR-MS(ESI)m / z 460.1(M+1).

[0479] The synthesis method of intermediate 126-3 is as follows:

[0480] Step 1: Add 115-2 (0.5 g, 2.64 mmol) and anhydrous THF (30 mL) to a round-bottom flask. Add NaH (60%, 211 mg) under ice bath conditions. After stirring for 30 minutes, add ethyl chloroformyl (430 mg, 3.96 mmol) and react at room temperature for 2 hours. After the reaction is complete, quench the reaction by adding saturated ammonium chloride solution dropwise under ice bath conditions. Extract with ethyl acetate, combine the organic phases and wash with saturated brine. After drying with anhydrous sodium sulfate, separate by column chromatography to obtain 142-1 (400 mg, yellow oil, yield 57.9%). 1 HNMR (400MHz, DMSO-d6) δ9.03 (s, 1H), 8.20 (s, 1H), 7.23 (d, J = 2.7Hz, 1H), 6.62 (dd, J = 3.4, 1.9 Hz, 1H), 4.36 (q, J=4.2Hz, 2H), 2.83 (s, 6H), 1.26 (t, J=4.3Hz, 3H). LR-MS (ESI) m / z388.1 (M+1).

[0481] Steps 2 and 3 can be obtained by referring to steps 3 and 4 of Example 115 to obtain 126-3 as a brown solid. 1HNMR (400MHz, DMSO-d6) δ 8.63 (s, 1H), 7.16 (s, 1H), 6.65 (s, 2H), 4.36 (q, J = 4.2Hz, 2H), 1.26 (t, J = 4.3Hz, 3H). LR-MS (ESI) m / z 333.1 (M+1).

[0482]

[0483] Example 127 Preparation of 3-((4-amino-7-(cyclopropanoyl)-7-H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-(thiophene-3-ylmethyl)benzamide

[0484] Except for replacing chloroformyl ethyl ester with chloroformylcyclopropane in step 1, the synthesis method is as described in Example 126. 1 H NMR(400MHz, DMSO-d6)δ8.96(t,J=6.0Hz,1H),8.13(s,1H),8.12(d,J=2.0Hz,1H) ,7.77(dd,J=7.6,1.9Hz,1H),7.37–7.29(m,1H),7.30(s,1H),7.22(dd,J=4.8,1. 8Hz,1H),7.06(t,J=1.6Hz,1H),7.04(dd,J=4.9,1.6Hz,1H),6.62(s,2H),4.55(d ,J=5.9Hz,2H),2.53(s,3H),1.92–1.80(m,1H),1.20–0.99(m,4H).LR-MS(ESI)m / z 456.1(M+1).

[0485] Example 128 Preparation of 3-((4-amino-7-isopropyl-7-H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-((5-methylthiophen-3-yl)methyl)benzamide

[0486] Except for replacing iodomethane with 2-iodopropane in step 1 and replacing 3-iodo-4-methyl-N-(5-methylthiophene-3-ylmethyl)benzamide with 3-iodo-4-methyl-N-(thiophene-3-ylmethyl)benzamide in step 3, the synthesis method is the same as in Example 72. 1H NMR (500MHz, chloroform-d) δ8.84(t,J=5.8Hz,1H),8.18(s,1H),7.87(d,J=1.9Hz,1H),7 .77(dd,J=8.1,2.0Hz,1H),7.55(s,1H),7.32(dd,J=8.1,1.0Hz,1H),6.90(d,J= 1.6Hz,1H),6.68(dd,J=1.6,0.7Hz,1H),5.96(s,2H),4.74–4.69(m,1H),4.51(d ,J=5.7Hz,2H),2.52(s,3H),2.36(s,3H),1.53(d,J=4.9Hz,6H).LR-MS(ESI)m / z 444.2(M+1).

[0487] Example 129 Preparation of 3-((4-amino-7-isopropyl-7-H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-N-((5-chlorothiophene-3-yl)methyl)-4-methylbenzamide

[0488] Except for replacing iodomethane with 2-iodopropane in step 1 and replacing 3-iodo-4-methyl-N-(5-chlorothiophene-3-ylmethyl)benzamide with 3-iodo-4-methyl-N-(thiophene-3-ylmethyl)benzamide in step 3, the synthesis method is the same as in Example 72. 1 H NMR (500MHz, DMSO-d6) δ8.92(t,J=5.9Hz,1H),8.19(s,1H),7.87(d,J=1.9Hz,1H),7.80(dd,J=8.1,1.9Hz,1H),7.55(s,1H),7.33(dd,J=8.1,1.0Hz,1H) ,7.03(d,J=1.8Hz,1H),6.87(d,J=1.6Hz,1H),6.58(s,2H),4.77–4.68(m,1 H),4.54(d,J=6.0Hz,2H),253(s,3H),1.50(d,J=4.9Hz,6H).LR-MS(ESI)m / z 464.1(M+1).

[0489] Example 130 Preparation of 3-((4-amino-7-isopropyl-7-H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-N-((5-methoxythiophene-2-yl)methyl)-4-methylbenzamide

[0490] Except for replacing iodomethane with 2-iodopropane in step 1 and replacing 3-iodo-4-methyl-N-(5-methoxythiophene-2-ylmethyl)benzamide with 3-iodo-4-methyl-N-(thiophene-3-ylmethyl)benzamide in step 3, the synthesis method is the same as in Example 72. 1 H NMR(500MHz,DMSO-d6)δ9.02(t,J=6.0Hz,1H),8.25(s,1H),7.94(d,J=1.9Hz ,1H),7.87(dd,J=8.2,1.9Hz,1H),7.62(s,1H),7.41–7.37(m,1H),6.97(d,J= 7.0Hz,1H),6.87(d,J=7.0Hz,1H),6.61(s,2H),4.85–4.77(m,1H),4.58(d,J =5.6Hz,2H),3.93(s,3H),2.52(s,3H),1.56(d,J=4.9Hz,6H).LR-MS(ESI)m / z 460.2(M+1).

[0491] Example 131 Preparation of 3-((4-amino-7-isopropyl-7-H-pyrrolo[2,3-d]pyrimidin-5-yl)ethynyl)-4-methyl-N-((5-methylthiophen-2-yl)methyl)benzamide

[0492] Except for replacing iodomethane with 2-iodopropane in step 1 and replacing 3-iodo-4-methyl-N-(5-methylthiophene-2-ylmethyl)benzamide with 3-iodo-4-methyl-N-(thiophene-3-ylmethyl)benzamide in step 3, the synthesis method is the same as in Example 72. 1 H NMR(400MHz, DMSO-d6)δ8.90(t,J=5.8Hz,1H),8.19(s,1H),7.87(d,J=1.9Hz,1H) ,7.80(dd,J=8.1,1.9Hz,1H),7.55(d,J=0.7Hz,1H),7.33(dd,J=8.1,1.0Hz,1H),6 .99(d,J=6.3Hz,1H),6.89(d,J=6.3Hz,1H),6.61(s,2H),4.79–4.69(m,1H),4.54 (d,J=5.7Hz,2H),2.52(s,3H),2.35(s,3H),1.50(d,J=4.9Hz,6H).LR-MS(ESI)m / z 444.2(M+1).

[0493] Example 13: Preparation of 2-ethyl(7-isopropyl-5-((2-methyl-5-((thiophen-3-ylmethyl)carbamoyl)phenyl)ethynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate

[0494]

[0495] Compound 77 (80 mg, 0.186 mmol), DIPEA (72 mg, 0.559 mmol), and anhydrous DMF (2 mL) were added to a round-bottom flask. Ethyl chloroformate (30 mg, 0.279 mmol) was added dropwise under ice bath conditions. After stirring for 1 hour, the reaction was quenched by adding methanol. The solvent was evaporated under reduced pressure, and the mixture was separated by column chromatography to give compound 132 (30 mg, yellow solid, yield: 32%). 1 H NMR (400MHz, chloroform-d) δ8.71(s,1H),8.42(s,1H),8.02(s,1H),7.74(d,J=8.1Hz,1H), 7.47(s,1H),7.37–7.29(m,2H),7.26–7.22(m,1H),7.11(d,J=5.0Hz,1H),6.63(br s,1H),5.12(p,J=6.6Hz,1H),4.67(d,J=5.5Hz,2H),4.29(q,J=7.0Hz,2H),2.58(s,3H),1.55(d,J=6.8Hz,6H),1.30(t,J=7.3Hz,3H).LR-MS(ESI)m / z 502.2(M+1).

[0496] Example 133 Preparation of isopropyl (7-isopropyl-5-((2-methyl-5-((thiophen-3-ylmethyl)carbamoyl)phenyl)ethynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate

[0497] Except for replacing ethyl chloroformate with isopropyl chloroformate, the synthesis method is as described in Example 132. 1H NMR(400MHz, DMSO-d6)δ9.92(s,1H),8.90(t,J=6.0Hz,1H),8.29(s,1H),7.87(d,J=1 .9Hz,1H),7.80(dd,J=8.1,1.9Hz,1H),7.57(s,1H),7.36–7.29(m,2H),7.24(t,J=1.7 Hz,1H),6.98(dd,J=5.0,1.7Hz,1H),5.03–4.92(m,1H),4.78–4.69(m,1H),4.55(d,J= 5.9Hz,2H),2.52(s,3H),1.50(d,J=4.9Hz,6H),1.29(d,J=5.7Hz,6H).LR-MS(ESI)m / z 516.2(M+1).

[0498] (II) Examples of Bioactivity Detection

[0499] Experimental Example 1: Inhibitory Activity of Compounds on the Proliferation of KIT Cells Carrying Different Drug-Resistant Mutations

[0500] Experimental methods:

[0501] The effect of the compound on cell proliferation was detected using the MTT assay. Healthy cells were collected and seeded into 96-well plates, with different concentrations of the compound added. The plates were incubated at 37°C with 5% CO2 and saturated humidity for 72 hours. After drug treatment, MTT was added to each well, and the plates were incubated at 37°C with 5% CO2 and saturated humidity for another 4 hours. A triple solution (10% SDS, 5% isobutanol, 0.01 mol / L HCl) was added, and the plates were incubated at 37°C for 12 hours to ensure complete dissolution of the blue-purple formazan. OD values ​​were measured at 570 nm and 690 nm using a microplate reader. The inhibitory rate of the compound on cell proliferation was calculated using the following formula:

[0502] Inhibition rate (%) = (OD value of control well - OD value of drug well) / OD value of control well × 100%

[0503] Calculation of compound IC using Graphpad Prism 8.0 software 50 And generate a concentration (logarithmic)-inhibition rate plot.

[0504] Experimental results:

[0505] The inhibitory activity of the compound and positive control drug Ripretinib on the proliferation of 32D KIT D816V cells in the embodiments of this invention is shown in Table 2:

[0506] Table 2. Inhibitory effects of compounds on the proliferation of 32D KIT D816V cells.

[0507]

[0508]

[0509]

[0510] Experimental Conclusions: As shown in Table 2, the heterocyclic alkyne compounds of the present invention all exhibited significant inhibitory activity against the proliferation of 32D KIT D816V cells, with 33 compounds showing superior activity against Ripretinib. Examples 76, 77, 99, and 120 show the IC50 values ​​for their inhibitory activity against the proliferation of 32D KIT D816V cells. 50 The value was less than 1 nM, which was significantly better than the positive control drug Ripretinib, showing that this class of compounds has a strong advantage in inhibiting the proliferation of cells carrying the KIT D816V drug-resistant mutation.

[0511] Representative compounds and the positive control drug Ripretinib were selected to further evaluate their inhibitory activity on the proliferation of other KIT mutant cells (32DKIT V559D, 32D KIT V559D-V654A, 32D KIT V559D-Y823D and 32D KIT V559D-N822K), KIT wild-type cells (NCI-H526, Mo7e, HMC-1), and 32D cells. The results are shown in Table 3.

[0512] Table 3 Comparison of the inhibitory activities of representative compounds on the proliferation of KIT cells with different mutations, wild-type KIT cells, and normal cells.

[0513]

[0514]

[0515] NT: Not test.

[0516] The results showed that the representative compounds exhibited potent inhibitory activity against the proliferation of cells carrying different mutant KIT cells, namely V559D, V559D-V654A, V559D-Y823D, and V559D-N822K, demonstrating the advantage of this class of compounds in broad-spectrum inhibition of mutant KIT. Simultaneously, the representative compounds showed weaker activity against wild-type KIT-dependent cell lines, but possessed a higher therapeutic index compared to mutant KIT, such as against KIT... D861V The selectivity index was higher than 490-fold, and superior to the positive control drug ripretinib. Furthermore, the compound did not exhibit cytotoxicity against 32D normal cells, demonstrating excellent selectivity and avoiding potential off-target toxicity.

[0517] Further testing was conducted on the inhibitory activity of representative compounds against the proliferation of other wild-type target-dependent cell lines, including PDGFRα (U118MG) and EGFR (A431), and the results are shown in Table 4.

[0518] The results showed that the representative compounds did not exhibit cytotoxicity against wild-type PDGFR and EGFR-dependent cell lines, demonstrating good target selectivity. However, the positive control drug ripretinib showed significant inhibitory activity against PDGFR-dependent U118MG cells, with a low therapeutic index.

[0519] Table 4. Inhibitory activities of representative compounds on the proliferation of PDGFRα and EGFR-dependent cell lines.

[0520]

[0521] Experimental Example 2: Effects of representative compounds 27, 48, 77, and 85 on KIT and its mediated downstream signaling

[0522] Western blotting was used to detect the effects of the compounds on KIT and its downstream signaling pathways in 32D KIT D816V, 32D KIT V559D, and 32D KIT V559D-V654A cells.

[0523] Experimental methods:

[0524] Cells were seeded in six-well plates, and different concentrations of drugs were added. The plates were then incubated at 37°C, 5% CO2, and saturated humidity for 4 hours. Cells were lysed on ice using 1×SDS gel loading buffer (50mM Tris-HCl (pH 6.8), 100mM DTT, 2% SDS, 10% glycerol, 0.1% bromophenol blue). Cell lysates were denatured by heating in a boiling water bath for 10 min and then subjected to SDS-PAGE electrophoresis. After electrophoresis, proteins were transferred to PVDF membranes and blocked in blocking buffer (5% skim milk diluted in TBST) at room temperature for 1 hour. After washing, the appropriate primary and secondary antibodies were added, and the membranes were developed using ECL reagent. Finally, the membranes were observed and photographed using an ECL chemiluminescence imaging system.

[0525] Experimental conclusion:

[0526] Representative compounds and the positive control drug Ripretinib inhibited KIT and its downstream signaling pathways in tumor cells carrying different mutant KIT. Results are shown in [link to relevant documentation]. Figure 1-3 .Depend on Figure 1-3 It is known that the compounds containing heterocyclic alkyne groups in the embodiments of the present invention can significantly inhibit the activation of different mutant forms of KIT and its downstream signaling pathways at the cellular level.

[0527] Experiment Example 3: In vivo drug efficacy evaluation test in mice

[0528] Experimental methods:

[0529] BALB / c-nu nude mice, 5-6 weeks old, female. Each nude mouse was subcutaneously inoculated with 32D KIT D816V cells until the average tumor volume reached ~100mm. 3 Subsequently, the animals were grouped according to tumor volume (D0). Mice were administered the drug by gavage (ig) twice daily at a volume of 10 mL / kg; the solvent group received the same volume of "solvent"; tumor volume was measured twice weekly, mouse weight was recorded, and data were recorded.

[0530] Compounds 77, 85, and Repritinib were all prepared using "solvents" (5% DMSO / 5% Etoh / 40% PEG 400 / 50% H2O).

[0531] Experimental results:

[0532] Compound 85 (30 and 100 mg / kg, ig, BID×12) dose-dependently inhibited the growth of subcutaneous xenografts in 32D KIT D816V nude mice, with inhibition rates of 20% and 50%, respectively. Compound 77 (20 and 40 mg / kg, ig, BID×12) also dose-dependently inhibited the growth of subcutaneous xenografts in 32D KIT D816V nude mice, with inhibition rates of 45% and 71%, respectively. Ripretinib (30 and 100 mg / kg, ig, BID×12) showed inhibition rates of 13% and 21% against subcutaneous xenografts in 32D KIT D816V nude mice, respectively. The tumor-bearing mice tolerated all the drugs well, without significant weight loss or other symptoms. Comparatively, compounds 77 and 85 were significantly more effective than ribretinib against subcutaneous xenografts in 32D KIT D816V nude mice (P<0.05, comparison of high-dose groups).

[0533] In summary, the compounds containing heterocyclic alkyne groups in the embodiments of the present invention exhibit strong inhibitory activity against 32D KIT D816V cells. Meanwhile, representative compounds 27, 48, 77, and 85 all demonstrate potent inhibitory activity against different mutant forms of KIT, including V559D, V559D-V654A, V559D / Y823D, and V559D / N822K, indicating that these compounds possess the advantage of potent and broad-spectrum inhibition of different types of mutant KIT. Representative compounds 27, 48, 77, and 85 showed weak inhibitory effects on normal 32D cells and cells carrying wild-type KIT, PDGRFα, and EGFR, indicating that these compounds have high selectivity and can avoid related off-target toxic side effects. Importantly, representative compounds 77 and 85 showed significant efficacy in the 32D KIT D816V nude mouse subcutaneous xenograft model; their efficacy was significantly stronger than that of the reference compound Ripretinib (P<0.05), indicating that the compounds containing heterocyclic alkyne groups have the advantage of stronger in vivo efficacy.

[0534] The preferred embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the specific details of the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and such simple modifications are all within the protection scope of the present invention.

Claims

1. A compound of formula (II), or a deuterated compound thereof, or a pharmaceutically acceptable salt thereof: in: R 1 Selected from halogens and C1-3 alkyl groups; R a R b Each is independently selected from hydrogen, deuterium, halogen, methyl; or R a and R b Together with the carbon atoms connected to them, they form a ternary carbon ring; Ring M 1 Selected from the following structures: The "C,N" on the ring atom indicates that the atom is CH or N. The R 2 Each group is independently selected from halogen, cyano, C1-6 alkyl, C1-6 heteroalkyl, C1-6 alkoxycarbonyl, C2-6 alkenyl, C2-6 alkynyl, C3-6 cycloalkyl, 3-6 membered heterocyclic alkyl, C4-8 cycloalkenyl, 4-8 membered heterocyclic alkenyl, C6-10 aryl, 5-7 membered heteroaryl, 5-7 membered heterocyclic, -C(=O)(C1-6 alkyl), -C(=O)(C3-6 cycloalkyl), -OR 3 -N(R) 3 2, -(C1-6 alkyl)OR 3 -C(=O)N(R) 3 )2、-(C1-6 alkyl)-C(=O)N(R 3 )2、-SR 3 -S(=O)R 3 -S(=O)2R 3 -N(R) 3 )C(=O)R 3 -(C1-6 alkyl)-N(R) 3 )C(=O)R 3 -N(R) 3 )S(=O)2R 3 -P(=O)(R 3 (R) 3 ); wherein the C1-6 alkyl, C1-6 heteroalkyl, C1-6 alkoxycarbonyl, C2-6 alkenyl, C2-6 alkynyl, C3-6 cycloalkyl, 3-6 heterocyclic alkyl, 4-8 heterocyclic alkenyl, 4-8 heterocyclic alkenyl, C6-10 aryl, 5-7 heteroaryl, 5-7 heterocyclic are optionally surrounded by 1-5 R 3 replace; The R 3 Each is independently selected from hydroxyl, cyano, amino, halogen, C1-6 alkyl, C1-6 alkoxycarbonyl, C3-C6 cycloalkyl, C1-6 haloalkyl, hydroxyC1-6 alkyl, -NH (C3-6 cycloalkyl), -NHC (=O) (C1-6 alkyl), and cyano-substituted 5-7 membered heterocyclic methyl groups selected from one or more of O, N, and S; R H1 and R H2 Each group is independently selected from hydrogen, halogen, amino, cyano, hydroxyl, -N(R) 4 (R) 5 ); The R 4 R 5 Each is independently selected from hydrogen, C1-6 alkyl, C1-6 heteroalkyl, C3-6 cycloalkyl, C3-6 heterocycloalkyl, C1-6 alkylacyl, and C3-6 cycloalkylacyl. R H3 Selected from hydrogen, unsubstituted or substituted C1-6 alkyl, unsubstituted or substituted C1-6 heteroalkyl, -C(=O)OC1-6 alkyl, -C(=O)OC3-6 cycloalkyl, -C(=O)C1-6 alkyl, -C(=O)C3-6 cycloalkyl, unsubstituted or substituted C3-6 cycloalkyl, unsubstituted or substituted C2-6 alkenyl, unsubstituted or substituted C4-6 cycloalkenyl, unsubstituted or substituted C6-10 aryl, unsubstituted or substituted 5-7 membered heteroaryl, and unsubstituted or substituted 4-7 membered heterocyclic groups; wherein the substitution refers to each being independently substituted by 1-5 R groups. 6 The substituted rings are: in cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic groups, one or more cyclic C atoms are optionally replaced by a corresponding number of C (=O) groups, and one or more cyclic S or N atoms are optionally oxidized to form S-oxides or N-oxides; The R 6 Each group is independently selected from halogen, cyano, C1-6 alkyl, C1-6 heteroalkyl, C2-6 alkenyl, C2-6 ynyl, C3-6 cycloalkyl, C4-8 cycloalkenyl, C6-10 aryl, 5-7 membered heteroaryl, 5-7 membered heterocyclic, -C(=O)R 7 OR 7 -N(R) 7 2, -(C1-6 alkyl)OR 7 -C(=O)N(R) 7 )2、-(C1-6 alkyl)-C(=O)N(R 7 )2、-SR 7 -S(=O)R 7 -S(=O)2R 7 -N(R) 7 )C(=O)R 7 -(C1-6 alkyl)-N(R) 7 )C(=O)R 7 -N(R) 7 )S(=O)2(R 7 -P(=O)(R) 7 (R) 7 ); wherein the C1-6 alkyl, C1-6 heteroalkyl, C2-6 alkenyl, C2-6 ynyl, C3-6 cycloalkyl, C4-8 cycloalkenyl, C6-10 aryl, 5-7 heteroaryl, and 5-7 heterocyclic groups are optionally surrounded by 1-5 R groups. 7 replace; The R 7 Each is independently selected from halogen, hydroxyl, cyano, amino, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-6 cycloalkyl, C1-6 heteroalkyl, C4-6 heterocycloalkyl, C1-6 haloalkyl, hydroxy C1-6 alkyl, NH (C1-6 cycloalkyl), -NHC (=O) (C1-6 alkyl), C (=O) (C1-6 alkyl), C (=O) (C3-6 cycloalkyl); Ring M 2 Selected from unsubstituted or substituted C6-10 aryl groups and unsubstituted or substituted 5-7 heteroaryl groups; wherein substitution refers to each being independently substituted by 1-5 R groups. M Replaced; The R M Each is independently selected from halogen, cyano, C1-6 heteroalkyl, C2-6 alkenyl, C2-6 ynyl, C3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C4-6 cycloalkenyl, C6-10 aryl, 5-7 membered heteroaryl, -C(=O)(C1-6 alkyl), -C(=O)(C3-6 cycloalkyl), -(C1-6 alkyl)-R 8 -OR 8 -N(R) 8 2, -NH(C1-6 alkyl)-R 8 -O(C1-6 alkyl)-R 8 -C(=O)N(R) 8 )2、-SR 8 -S(=O)R 8 -S(=O)2R 8 The C1-6 alkyl, C3-6 cycloalkyl, 3-6 heterocyclic alkyl, C4-6 cycloalkenyl, 4-6 heterocyclic alkenyl, C6-10 aryl, and 5-7 heteroaryl groups are each optionally independently bound by 1-5 R groups. 8 replace; The R 8 Each is independently selected from hydroxyl, cyano, amino, halogen, -NH (C1-6 alkyl), C1-6 alkyl, C1-6 heteroalkyl, C1-6 haloalkyl, C1-6 hydroxyalkyl, C3-6 cycloalkyl, and C3-6 heterocycloalkyl.

2. The compound according to claim 1, or its deuterated compound, or a pharmaceutically acceptable salt thereof, wherein... R 1 Selected from methyl, F, and Cl.

3. The compound according to claim 1, or its deuterated compound, or a pharmaceutically acceptable salt thereof, wherein... Ring M 2 Selected from unsubstituted or substituted phenyl, unsubstituted or substituted thiophene, unsubstituted or substituted pyrazolyl, unsubstituted or substituted thiazolyl, unsubstituted or substituted pyridyl, unsubstituted or substituted oxazolyl, unsubstituted or substituted isoxazolyl, unsubstituted or substituted benzothiophene; wherein the substitution refers to each being independently substituted by 1-5 R... M Replaced; The R M Each is independently selected from halogen, cyano, C1-6 alkyl, C1-6 heteroalkyl, C2-6 alkenyl, C2-6 ynyl, C3-6 cycloalkyl, 3-6 membered heterocyclic alkyl, C4-6 cycloalkenyl, 4-6 membered heterocyclic alkenyl, C6-10 aryl, 5-7 membered heteroaryl, -C(=O)(C1-6 alkyl), -C(=O)(C3-6 cycloalkyl), -(C1-6 alkyl)-R 8 -C(=O)N(R) 8 )2、-SR 8 -S(=O)R 8 -S(=O)2R 8 The C1-6 alkyl, C3-6 cycloalkyl, 3-6 heterocyclic alkyl, C4-6 cycloalkenyl, 4-6 heterocyclic alkenyl, C6-10 aryl, and 5-7 heteroaryl groups are each optionally independently bound by 1-5 R groups. 8 replace; The R 8 Each is independently selected from hydroxyl, cyano, amino, halogen, -NH (C1-6 alkyl), C1-6 alkyl, C1-6 heteroalkyl, C1-6 haloalkyl, C1-6 hydroxyalkyl, C3-6 cycloalkyl, and C3-6 heterocycloalkyl.

4. A compound, or a deuterated compound thereof, or a pharmaceutically acceptable salt thereof, wherein, The compound is selected from the following compounds:

5. A method for preparing the compound of claim 1, comprising the following steps: Compound (A) undergoes a coupling reaction with compound (B) to yield compound (II): in, Ring M 1 Ring M 2 X and R 1 The definition is as described in claim 1; X is -C(R) a (R) b )-;TMS is -Si(CH3)3.

6. The method according to claim 5, wherein The coupling reaction was carried out in a solvent in the presence of a base in the presence of palladium and copper metal catalysts. The palladium metal catalyst is one or more of Pd(PPh3)2Cl2, Pd(OAc)2, and Pd(PPh3)4; The copper metal catalyst is CuI and / or CuCl; The alkali is CsF, Cs2CO3, KF, K2CO3, NaHCO3, Na2CO3, Et3N, ( i One or more of Pr)2EtN and DMAP; The solvent is one or more of acetonitrile, 1,4-dioxane, and DMF.

7. A pharmaceutical composition comprising one or more selected from the compounds according to any one of claims 1-4, their deuterated compounds, pharmaceutically acceptable salts, and pharmaceutically acceptable excipients.

8. Use of the compound according to any one of claims 1-4, its deuterated compound, a pharmaceutically acceptable salt, or the pharmaceutical composition according to claim 7 in the preparation of a KIT inhibitor.

9. Use of the compound of any one of claims 1-4 or its deuterated compound, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 7 in the preparation of a medicament for treating, preventing, or improving one or more diseases or conditions selected from tumors, inflammation, autoimmune diseases, and nervous system diseases.

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