Thiophene compound serving as cyclin-dependent kinase inhibitor as well as preparation method and medical application of thiophene compound

Abstract

The invention relates to a thiophene compound as a cyclin-dependent kinase inhibitor as well as a preparation method and medical application of the thiophene compound. Specifically, the invention relates to a compound as shown in a general formula (I), a preparation method thereof, a pharmaceutical composition containing the compound, and application of the compound as a cyclin dependent kinase (CDK) inhibitor in drugs for preventing or treating abnormal cell growth such as cancer. Wherein the definition of each group in the general formula (I) is the same as that in the specification. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and specifically relates to a heterocyclic compound, a preparation method thereof, a pharmaceutical composition containing the compound, and its use as a cyclin-dependent kinase (CDK) inhibitor in a drug for treating abnormal cell growth such as cancer. The compound of the present invention can be used to treat cancer. Background Art

[0002] Cyclin-dependent kinases (CDKs) belong to the serine / threonine protein kinase family and are key proteins that regulate the cell cycle. Currently, the discovered CDKs mainly include CDK1-15, etc., which play a role by binding to the corresponding cyclins to form a stable form of CDK / cyclin complex. In addition, cells contain endogenous CDKs and CDK / cyclin complex inhibitors (CKIs), which together form a regulatory network for the cell cycle and are strictly controlled during cell division. In many cancer cells, it has been found that the expression of CDKs, cyclins, and CKIs is out of control, and the CDK / cyclin complex is usually overexpressed.

[0003] The structures and functions of CDK4 and CDK6 are very similar, and both can bind to the three subtypes of cyclin D (cyclinD1 / D2 / D3) to form a complex, phosphorylating a series of substrates including retinoblastoma protein (Rb), thereby promoting the progression of the cell cycle. Phosphorylated Rb protein releases transcription factors such as E2F that bind to it, and E2F activates and transcribes a series of genes, causing cells to enter the S phase and start DNA replication (Kato et al., Genes Dev, 1993, 7(3), 331-342; Dyson et al., Genes Dev, 1998, 12(15), 2245-2262). The activity of CDK4 / 6 is also negatively regulated by the INK4 family. As an endogenous inhibitory protein, INK4 can competitively bind to CDK and inhibit the formation of the cyclin D-CDK4 / 6 complex (Baker et al., Genes & Cancer, 2012, 3(11-12), 658-669).

[0004] However, it has been found in most human cancers that the CyclinD-CDK4 / 6-p16-Rb pathway is abnormally and continuously activated, which can promote rapid development of the G1 phase and lead to abnormal cell proliferation. The main reasons include overexpression of cyclin D1 caused by gene rearrangement or gene amplification (Bergsagel et al., Blood, 2005, 106 (1), 296-303); p16INK4a gene deletion, point mutation, or DNA methylation leading to p16INK4a inactivation (Ruas et al., Biochim Biophys Acta, 1998, 1378 (2), F115-177); CDK4 / 6 gene amplification or point mutation (Hamilton et al., Cancer Treatment Reviews, 2016, 45, 129-138). Targeted intervention based on the abnormality of this pathway makes CDK4 / 6 one of the popular anti-tumor targets.

[0005] Among the CDK-targeted therapies, the most advanced are selective CDK4 / 6 inhibitors. The first-generation broad-spectrum CDK inhibitor, Flavopiridol (Alvocidib), can simultaneously inhibit CDK1 / 2 / 4 / 6 / 7 / 9, but due to its severe toxic side effects, it causes adverse reactions such as fatigue, diarrhea, and bone marrow suppression, and ultimately failed to be successfully applied in clinical practice (Senderowicz et al., Journal of Clinical Oncology, 1998, 16 (9), 2986-2999). Currently, three compounds, Palbociclib, Ribociclib, and Abemaciclib, have been approved by the FDA for first-line or second-line treatment of advanced breast cancer, and another drug, Lerociclib, is in clinical development.

[0006] Palbociclib is the first FDA-approved CDK4 / 6 inhibitor. It is an oral pyridine compound that inhibits the proliferation of ER+ breast cancer cells by reducing the phosphorylation of Rb protein, preventing the cell cycle from entering the S phase from the G1 phase. Several large-scale, randomized, prospective clinical studies have confirmed that Palbociclib combined with letrozole or fulvestrant can significantly prolong the patient's progression-free survival and has reliable safety (Cristofanilli et al., Lancet Oncol, 2016, 17(4), 425-439; Richard et al., Lancet Oncology, 2015; Finn et al., N Engl J Med, 2016, 375(20), 1925-1936; Nicholas et al., N Engl J Med, 2015, 373(3), 209-219). These clinical trial data support palbociclib in combination with an aromatase inhibitor or fulvestrant as standard of care for first-line treatment of women with ER+ / HER2- premenopausal or postmenopausal metastatic breast cancer.

[0007] Ribociclib (Kisqali, LEE011) and Abemaciclib (Verzenio, LY2835219) are both oral reversible CDK4 / 6 inhibitors that can cause G1 arrest by inhibiting Rb phosphorylation. IC of ribociclib on CDK4 / 6 inhibition 50 values ​​were 10 nM and 39 nM, respectively, while other CDK family members were less sensitive (IC values ​​for CDK1 and CDK2 were 50 The values ​​are all greater than 50mM) (Sherr et al., Cancer Discovery, 2016, 6(4), 353-367). After oral administration for 1-4 hours, the Cmax value can be reached in the plasma; after continuous administration for 8 days, the blood steady state is reached. Ribociclib has a binding rate of 70% to human plasma proteins and can be decomposed by CPY3A4 (a weak inhibitor) in the body. The main products are N-hydroxy and N-demethyl derivatives. Abemaciclib has higher selectivity for CDK4 / 6 kinases, and IC 50 The values ​​were 2 nM and 5 nM, respectively, while the IC values ​​for CDK1 and CDK2 inhibition were 50Value>500nM. Abemaciclib is a phenylpyrimidine compound, which is structurally similar to Palbociclib and Ribociclib. Abemaciclib exhibits a wider range of inhibitory effects and can also inhibit the Dyrk, PIM, HIPK and CAMK kinase families (Ki<10nM) (Chen et al., Molecular Cancer Therapeutics, 2016, 15(10), 2273-2281). In vivo pharmacokinetic data showed that the peak value was reached within 4-24 hours after oral administration and steady state was reached after 5 days of continuous administration. Abemaciclib has a binding rate of approximately 96% to human plasma proteins and can be decomposed by CPY3A4 in the body. The main decomposition product is an N-deethyl derivative. The introduction of deuterium atoms gives the compound stronger metabolic stability, which is increased by 11-45%, which is manifested as a higher half-life. The drug is approved for combination with Fluvestrant for the treatment of ER+ / HER2- advanced or metastatic breast cancer. In addition, in xenograft tumors, Abemaciclib combined with gemcitabine also showed synergistic anti-tumor activity and had inhibitory effects on multiple types of tumors, including mantle cell lymphoma (MCL), colorectal cancer, lung cancer, glioblastoma and acute myeloid leukemia (AML).

[0008] However, treatment with CDK4 / 6 inhibitors may produce adverse effects, such as gastrointestinal or hematological toxicity, and acquired resistance may develop over time. Emerging data suggest that cyclin D3-CDK6 may be involved in the observed hematological toxicity (Malumbres et al., Mammalian Cells Cycle without the D-type Cyclin-Dependent Kinases Cdk4 and Cdk6, (2004) Cell 118 (4): 493-504; Sicinska et al., Essential Role for Cyclin D3 in Granulocyte Colony Stimulating Factor-Driven Expansion of Neutrophil Granulocytes (2006), Mol. Cell Biol 26 (21): 8052-8060; Cooper et al., A unique function for cyclin D3 in early B cell development, (2006), Nat. Immunol. 5 (7): 489-497). CDK4 has been identified as a single oncogenic driver in many breast cancers. CDK4 selective inhibitors may offer an improved safety profile or enhanced overall efficacy due to the potential for higher dosing compared to dual CDK4 / 6 inhibitors.

[0009] Therefore, there is still a need for improved therapies for cancer treatment. The compounds, compositions and methods of the present invention have one or more advantages, such as greater efficacy; potential for reduced side effects; potential for reduced drug-drug interactions; potential for improved dosing schedules; or potential for overcoming resistance mechanisms, etc. Summary of the invention

[0010] After intensive research, the present inventors have designed and synthesized a series of heterocyclic compounds, which show the ability to inhibit cyclin-dependent kinase (CDK) activity and can be developed as drugs for treating or preventing diseases associated with CDK activity, such as cancer.

[0011] The present invention provides a compound represented by general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

[0012]

[0013] in,

[0014] Ring A is a 5-6 membered heteroaryl group;

[0015] X is CR 7 or N;

[0016] Y is CR 8 or N;

[0017] Z is CR 9 or N;

[0018] Q is NR 10 , O or CR 11 R 12 , where R 11 and R 12 Together with the C atoms to which they are attached, they form a 13 or a 4-6 membered heterocyclic ring with O as a ring member;

[0019] Each R 3 independently selected from hydrogen, deuterium, halogen, oxo, hydroxyl, cyano, nitro, alkyl, alkoxy, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, alkylthioether, -NR a R b 、-C(O)R c 、-OC(O)R c 、-C(O)OR c 、-C(O)NR a R b 、-S(O) p R c 、-S(O) p NR a R b 、-S(O)(=NR a )R c 、-SF 5 、-P(O)R a R b 、-P(O)(OR c )(OR d ) and -B(OR c )(OR d ), wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted by one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

[0020] R 4 Selected from hydrogen atoms, halogens, -NR a R b, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

[0021] R 5 Selected from hydrogen atoms, halogens, -NR a R b , nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

[0022] R 6 Selected from hydrogen atoms, halogens, -NR a R b , nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

[0023] R 7 , R 8 and R 9 are each independently selected from hydrogen, halogen, -NR a R b , nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

[0024] R 10 and R 13are independently selected from hydrogen, alkyl, halogenated alkyl, -S(O) p R c 、-S(O) p NR a R b 、-C(O)R a 、-C(O)OR a and -C(O)NR a R b , the alkyl and haloalkyl are each independently optionally further selected from halogen, -NR a R b , nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroaryl, -S(O) p R c 、-S(O) p NR a R b 、-C(O)R a 、-C(O)OR a 、-C(O)NR a R b and -OC(O)R a is substituted by one or more groups;

[0025] R a , R b , R c and R d each independently selected from hydrogen, halogen, hydroxy, cyano, amino, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxy, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

[0026] Or, R a and R b Together with the nitrogen atom to which they are attached, they form a heterocyclic group, which is optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl;

[0027] p is 1 or 2;

[0028] q is 1, 2, 3 or 4.

[0029] In a preferred embodiment, the compound represented by general formula (I) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt is a compound represented by general formula (II) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt,

[0030]

[0031] Among them, ring A, R 3 , R 4 , R 5 , R 6 , R 7 , Q and q are as defined in the general formula (I).

[0032] In a preferred embodiment, the compound represented by general formula (I) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt is a compound represented by general formula (III) or general formula (IV) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt,

[0033]

[0034] Among them, ring A, R 3 , R 4 , R 5 , R 6 , R 7 , R 10 and q are as defined in the general formula (I).

[0035] In a preferred embodiment, the compound represented by general formula (I) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt is a compound represented by general formula (V) or general formula (VI) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt,

[0036]

[0037] Among them, ring A, R 3 , R 4 , R 5 , R 7 and q are as defined in the general formula (I).

[0038] In a preferred embodiment, the compound represented by the general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein ring A is selected from pyrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, triazolyl, tetrazolyl, pyrrolyl, furanyl, thienyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl and triazinyl, preferably pyrazolyl, imidazolyl and pyridinyl.

[0039] In a preferred embodiment, the compound represented by the general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein each R 3 independently selected from hydrogen atoms, deuterium atoms, halogens, oxo groups, hydroxyl groups, cyano groups, nitro groups, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl, preferably hydrogen, halogen, oxo, hydroxyl, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-6 Cycloalkyl.

[0040] In a preferred embodiment, the compound represented by general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein q is 1 or 2.

[0041] In a preferred embodiment, the compound represented by the general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein:

[0042] Selected from

[0043]

[0044] In a preferred embodiment, the compound represented by the general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 4 Selected from hydrogen atoms, halogens, -NH 2 , nitro, cyano, hydroxyl, thiol, C 1-6 Alkyl, C1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl, preferably hydrogen atom, halogen, -NH 2 , hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 Hydroxyalkyl, more preferably hydrogen atom, halogen, C 1-6 Alkyl and C 1-6 Halogenated alkyl.

[0045] In a preferred embodiment, the compound represented by the general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 5 Selected from hydrogen atoms, halogens, -NH 2 , nitro, cyano, hydroxyl, thiol, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl, preferably hydrogen atom, halogen, -NH 2 , hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 Hydroxyalkyl, more preferably halogen and C 1-6 alkyl.

[0046] In a preferred embodiment, the compound represented by the general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 7 Selected from hydrogen atoms, halogens, -NH 2 , nitro, cyano, hydroxyl, thiol, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl, preferably hydrogen atom, halogen, -NH 2 , hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 A hydroxyalkyl group is more preferably a hydrogen atom.

[0047] In a preferred embodiment, the compound represented by the general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 6 Selected from hydrogen atoms, halogens, -NH 2 , nitro, cyano, hydroxyl, thiol, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl, preferably hydrogen atom, halogen, -NH 2 , hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 A hydroxyalkyl group is more preferably a hydroxy group.

[0048] In a preferred embodiment, the compound represented by the general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein

[0049] R a and R b are each independently selected from hydrogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, 3-6 membered heterocyclic group, wherein the C 1-6 Alkyl, C 3-8 The cycloalkyl and 3-6 membered heterocyclic groups are each independently further selected from halogen, amino, cyano, hydroxyl, mercapto, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 One or more groups in the haloalkoxy group are substituted;

[0050] Or, R a and R b Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group is optionally further selected from halogen, amino, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 One or more groups in the haloalkoxy group are substituted.

[0051] Typical compounds of the present invention include, but are not limited to:

[0052]

[0053]

[0054]

[0055]

[0056] or a stereoisomer, tautomer, meso racemate, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof.

[0057] The present invention also provides a method for preparing the compound represented by the general formula (I) according to the present invention or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, which comprises the following steps:

[0058]

[0059] In a solvent, in the presence of a base, compound Id reacts with compound Ie to obtain a compound represented by general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof;

[0060] The solvent is, for example, N-methylpyrrolidone, and the base is, for example, N,N-diisopropylethylamine;

[0061] Among them, rings A, X, Y, Z, Q, R 3 , R 4 , R 5 , R 6 and q are as defined in the general formula (I).

[0062] The present invention also provides a pharmaceutical composition comprising the compound according to the present invention or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0063] The present invention also provides the use of the compound according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or a pharmaceutical composition containing the same in the preparation of a cyclin-dependent kinase (CDK) inhibitor.

[0064] The present invention also provides the use of the compound according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or its pharmaceutically acceptable salt or a pharmaceutical composition containing the same in the preparation of a drug for inhibiting cancer cell proliferation, inhibiting cancer cell invasion or inducing cancer cell apoptosis.

[0065] The present invention also provides the use of the compound according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or a pharmaceutical composition containing the same in the preparation of a medicament for preventing and / or treating diseases associated with cyclin-dependent kinase (CDK) activity, such as cancer, in particular cancer characterized by amplification or overexpression of CDK4 and cyclin D3.

[0066] The present invention also relates to the compound according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or pharmaceutical composition comprising the same, which is used as a CDK inhibitor.

[0067] The present invention also relates to the compound according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or pharmaceutical composition comprising the same, for use in inhibiting CDK.

[0068] The present invention also relates to the compound according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or pharmaceutical composition containing the same, which is used for inhibiting cancer cell proliferation, inhibiting cancer cell invasion or inducing cancer cell apoptosis.

[0069] The present invention also relates to the compound according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or pharmaceutical composition comprising the same, for use in preventing and / or treating diseases associated with cyclin-dependent kinase (CDK) activity, such as cancer, in particular cancer characterized by amplification or overexpression of CDK4 and cyclin D3.

[0070] The present invention also relates to a method for inhibiting CDK, which comprises administering to a subject in need thereof an effective amount of a compound according to the present invention or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0071] The present invention also relates to a method for inhibiting cancer cell proliferation, inhibiting cancer cell invasion or inducing cancer cell apoptosis, which comprises administering to a subject in need thereof an effective amount of the compound according to the present invention or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0072] The present invention also relates to a method for preventing and / or treating a disease associated with cyclin-dependent kinase (CDK) activity, comprising administering to a subject in need thereof an effective amount of a compound according to the present invention or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, wherein the disease is, for example, cancer, in particular, cancer characterized by amplification or overexpression of CDK4 and cyclin D3.

[0073] In a preferred embodiment, the cancer according to the present invention is selected from breast cancer.

[0074] In an embodiment, the compound according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or pharmaceutical composition comprising the same can be administered or used in combination with another anti-cancer therapeutic agent or anti-cancer treatment method simultaneously, separately or sequentially.

[0075] According to the conventional methods in the field of the present invention, the compounds of the present invention can form pharmaceutically acceptable basic addition salts or acid addition salts with bases or acids. The bases include inorganic bases and organic bases, and acceptable organic bases include diethanolamine, ethanolamine, N-methylglucamine, triethanolamine, tromethamine, etc., and acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate and sodium hydroxide, etc. The acid includes inorganic acid and organic acid, and acceptable inorganic acid includes hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, etc. Acceptable organic acids include acetic acid, trifluoroacetic acid, formic acid, ascorbic acid, etc.

[0076] The pharmaceutical composition containing the active ingredient may be in a form suitable for oral administration, such as tablets, troches, lozenges, aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Oral compositions may be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more ingredients selected from the following: sweeteners, flavoring agents, colorants and preservatives to provide pleasing and palatable pharmaceutical preparations. Tablets contain the active ingredient and non-toxic pharmaceutically acceptable excipients suitable for preparing tablets for mixing. These excipients may be inert excipients such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating agents and disintegrants such as microcrystalline cellulose, crosslinked sodium carboxymethyl cellulose, corn starch or alginic acid; binders such as starch, gelatin, polyvinyl pyrrolidone or gum arabic; and lubricants such as magnesium stearate, stearic acid or talc. These tablets may be uncoated or may be coated by known techniques which mask the taste of the drug or delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained release action over a longer period of time. For example, water soluble taste masking materials such as hydroxypropylmethylcellulose or hydroxypropylcellulose, or time extending materials such as ethylcellulose, cellulose acetate butyrate may be used.

[0077] Oral preparations may also be provided in hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or in soft gelatin capsules wherein the active ingredient is mixed with a water-soluble carrier such as polyethylene glycol or an oily vehicle such as peanut oil, liquid paraffin or olive oil.

[0078] Aqueous suspensions contain the active substance and excipients suitable for preparing aqueous suspensions for mixing. Such excipients are suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone and gum arabic; dispersants or wetting agents, which may be naturally occurring phosphatides such as lecithin, or condensation products of alkylene oxides with fatty acids, for example, polyoxyethylene stearate, or condensation products of ethylene oxide with long chain fatty alcohols, for example, heptadecaethyleneoxy cetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol, for example, polyethylene oxide sorbitan monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example, polyethylene oxide dehydrated sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, for example ethylparaben or n-propylparaben, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, for example sucrose, saccharin or aspartame.

[0079] Oil suspensions can be prepared by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. The oil suspension may contain a thickener such as beeswax, hard paraffin or cetyl alcohol. The above-mentioned sweeteners and flavoring agents may be added to provide a palatable preparation. These compositions may be preserved by adding an antioxidant such as butylated hydroxyanisole or alpha-tocopherol.

[0080] Dispersible powders and granules suitable for preparing aqueous suspensions can provide the active ingredient and a dispersant or wetting agent, a suspending agent or one or more preservatives for mixing by adding water. Suitable dispersants or wetting agents and suspending agents are as described above. Other excipients such as sweeteners, flavoring agents and coloring agents can also be added. These compositions can be preserved by adding antioxidants such as ascorbic acid.

[0081] The pharmaceutical composition of the present invention can also be in the form of an oil-in-water emulsion. The oil phase can be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin or a mixture thereof. Suitable emulsifiers can be naturally occurring phospholipids, such as soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of the partial esters and ethylene oxide, such as polyethylene oxide sorbitol monooleate. Emulsions can also contain sweeteners, flavoring agents, preservatives, and antioxidants. Syrups and elixirs prepared with sweeteners such as glycerol, propylene glycol, sorbitol, or sucrose can be used. Such preparations can also contain a demulcent, a preservative, a coloring agent, and an antioxidant.

[0082] The pharmaceutical composition of the present invention may be in the form of a sterile injectable aqueous solution. Acceptable vehicles and solvents that may be used are water, Ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in an oil phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. The oil solution is then added to a mixture of water and glycerol and processed to form a microemulsion. The injection or microemulsion may be injected into the patient's bloodstream by local mass injection. Alternatively, the solution and microemulsion may be preferably administered in a manner that maintains a constant circulating concentration of the compound of the present invention. To maintain this constant concentration, a continuous intravenous drug delivery device may be used.

[0083] The pharmaceutical composition of the present invention can be in the form of a sterile injection water or oil suspension for intramuscular and subcutaneous administration. The suspension can be prepared according to known techniques with the above-mentioned suitable dispersants or wetting agents and suspending agents. The sterile injection preparation can also be a sterile injection solution or suspension prepared in a non-toxic parenterally acceptable diluent or solvent, such as a solution prepared in 1,3-butanediol. In addition, sterile fixed oils can be conveniently used as solvents or suspension media. For this purpose, any blended fixed oil including synthetic mono- or diglycerides can be used. In addition, fatty acids such as oleic acid can also be used to prepare injections.

[0084] The compounds of the invention may be administered in the form of suppositories for rectal administration. These pharmaceutical compositions may be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid in the rectum and which will dissolve in the rectum to release the drug. Such materials include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights and mixtures of fatty acid esters of polyethylene glycol.

[0085] It is well known to those skilled in the art that the dosage of a drug depends on a variety of factors, including but not limited to the following factors: the activity of the specific compound used, the patient's age, the patient's weight, the patient's health condition, the patient's behavior, the patient's diet, the administration time, the administration method, the excretion rate, the combination of drugs, etc. In addition, the best treatment method, such as the mode of treatment, the daily dosage of the general compound or the type of pharmaceutically acceptable salt can be verified according to traditional treatment plans.

[0086] The present invention may contain a compound and a pharmaceutically acceptable salt, hydrate or solvate thereof as an active ingredient, mixed with a pharmaceutically acceptable carrier or excipient to prepare a composition, and prepared into a clinically acceptable dosage form. The derivatives of the present invention may be used in combination with other active ingredients, as long as they do not produce other adverse effects, such as allergic reactions, etc. The compounds of the present invention may be used as the sole active ingredient, or in combination with other drugs for treating diseases associated with (CDK) activity. Combination therapy is achieved by administering the individual therapeutic components simultaneously, separately or sequentially.

[0087] Terminology

[0088] Unless stated otherwise, the terms used in the specification and claims have the following meanings.

[0089] The carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention include their isotopes, that is, the carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention are optionally further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include 12 C. 13 C and 14 C, hydrogen isotopes include protium (H), deuterium (D, also known as heavy hydrogen), tritium (T, also known as super tritium), oxygen isotopes include 16 O. 17 O and 18 O, sulfur isotopes include 32 S. 33 S. 34 S and 36 S, isotopes of nitrogen include 14 N and 15 N, fluorine isotopes include19 F, chlorine isotopes include 35 Cl and 37 Isotopes of Cl and bromine include 79 Br and 81 Br.

[0090] "Alkyl" refers to a saturated monovalent aliphatic hydrocarbon group, including straight and branched chain groups having a specified number of carbon atoms. Alkyl groups generally contain 1 to 20 carbon atoms (C 1 -C 20 Alkyl), preferably 1 to 12 carbon atoms (C 1 -C 12 Alkyl), more preferably 1 to 8 carbon atoms (C 1 -C 8 Alkyl) or 1 to 6 carbon atoms (C 1 -C 6 alkyl") or 1 to 4 carbon atoms (C 1 -C 4 Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2 ,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof, etc. The alkyl group may be substituted or unsubstituted, and when substituted, the substituents may be substituted at any available point of attachment.

[0091] The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, preferably an alkenyl group containing 2 to 6 carbon atoms, more preferably an alkenyl group containing 2 to 4 carbon atoms, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc. The alkenyl group may be substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

[0092] The term "alkynyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, preferably an alkynyl group containing 2 to 6 carbon atoms, more preferably an alkynyl group containing 2 to 4 carbon atoms or more preferably an alkynyl group containing 3 to 4 carbon atoms, such as ethynyl, propynyl, butynyl, etc. Alkynyl groups may be substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

[0093] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 8 carbon atoms, and even more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyls include spirocyclic, fused and bridged cycloalkyls.

[0094] The term "spirocycloalkyl" refers to a polycyclic group that shares a carbon atom (called a spiral atom) between 5 to 20 monocyclic rings, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiral atoms shared between the rings, the spirocycloalkyl is divided into a single spiral cycloalkyl, a double spiral cycloalkyl or a multi-spirocycloalkyl, preferably a single spiral cycloalkyl and a double spiral cycloalkyl. More preferably, it is a 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiral cycloalkyl. Non-limiting examples of spirocycloalkyl include:

[0095] The term "condensed cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with other rings in the system, wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic condensed cycloalkyl, preferably a bicyclic or tricyclic, more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl. Non-limiting examples of condensed cycloalkyls include:

[0096]

[0097] The term "bridged cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 members, and more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl, preferably a bicyclic, tricyclic or tetracyclic, and more preferably a bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl include:

[0098]

[0099] The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, etc. The cycloalkyl may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0100] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O). m(wherein m is an integer from 0 to 2) heteroatoms, but excluding the ring part of -OO-, -OS- or -SS-, the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; most preferably, it contains 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; more preferably, it contains 5 to 7 ring atoms, of which 1 to 2 or 1 to 3 are heteroatoms; even more preferably, it contains 4 to 6 ring atoms, of which 1 to 2 or 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably 1, 2, 5-oxadiazolyl, pyranyl or morpholinyl. Polycyclic heterocyclic groups include spirocyclic, fused ring and bridged heterocyclic groups.

[0101] The term "spiro heterocyclyl" refers to a polycyclic heterocyclic group in which the monocyclic rings of 5 to 20 members share one atom (called a spiro atom), wherein one or more of the ring atoms is selected from nitrogen, oxygen or S(O) m (wherein m is an integer 0 to 2) heteroatom, and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of shared spiral atoms between rings, the spiral heterocyclic group is divided into a single spiral heterocyclic group, a double spiral heterocyclic group or a multi-spiro heterocyclic group, preferably a single spiral heterocyclic group and a double spiral heterocyclic group. More preferably, it is a 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiral heterocyclic group. Non-limiting examples of spiral heterocyclic groups include:

[0102] The term "fused heterocyclyl" refers to a polycyclic heterocyclic group of 5 to 20 members, each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, and the remaining ring atoms are carbon. Preferably 6 to 14 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic, more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclic groups include:

[0103] The term "bridged heterocyclyl" refers to a 5- to 14-membered polycyclic heterocyclic group in which any two rings share two atoms that are not directly connected, which may contain one or more double bonds, but none of the rings has a completely conjugated π-electron system, and one or more of the ring atoms is selected from nitrogen, oxygen or S(O)m (wherein m is an integer from 0 to 2) heteroatoms, and the remaining ring atoms are carbon. Preferably, it is 6 to 14 members, and more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

[0104] The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclyl, non-limiting examples of which include: and wait.

[0105] The heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0106] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. More preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, non-limiting examples of which include:

[0107] The aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0108] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 5 to 10 members, containing 1 to 3 heteroatoms; more preferably 5 or 6 members, containing 1 to 3 or 1 to 2 heteroatoms; preferably, for example, imidazolyl, furanyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidyl, thiadiazole, pyrazinyl, etc., preferably imidazolyl, thiazolyl, pyrazolyl or pyrimidyl, thiazolyl; more preferably pyrazolyl or thiazolyl. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0109] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0110] The term "alkoxy" refers to-O-(alkyl) and-O-(cycloalkyl), wherein the definitions of alkyl and cycloalkyl are as described above. The non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0111] The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

[0112] The term "hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxy groups, wherein alkyl is as defined above.

[0113] The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.

[0114] The term "hydroxy" refers to an -OH group.

[0115] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0116] The term "amino" refers to -NH 2 .

[0117] The term "cyano" refers to -CN.

[0118] The term "nitro" refers to -NO 2 .

[0119] The term "oxo" refers to =0.

[0120] The term "carboxy" refers to -C(O)OH.

[0121] The term "thiol" refers to -SH.

[0122] The term "ester group" refers to -C(O)O(alkyl) or -C(O)O(cycloalkyl), wherein alkyl and cycloalkyl are as defined above.

[0123] The term "acyl" refers to a compound containing a -C(O)R group, wherein R is alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

[0124] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0125] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are replaced independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the skilled person can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxy groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (e.g. olefinic) bonds.

[0126] "Pharmaceutical composition" means a mixture containing one or more compounds described herein or their physiologically / pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

[0127] "Pharmaceutically acceptable salts" refer to salts of the compounds of the present invention, which are safe and effective when used in mammals and have the desired biological activity.

[0128] "Carrier" refers to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0129] "Cancer" refers to any malignant and / or aggressive growth or tumor (caused by abnormal cell growth). Cancers include solid tumors named after the cell type that formed them, cancers of the blood, bone marrow, or lymphatic system. Examples of solid tumors include sarcomas and carcinomas. Blood cancers include, but are not limited to, leukemias, lymphomas, and myelomas. Cancer also includes primary cancers that originate in a specific part of the body, metastatic cancers that have spread from the site where it started to other parts of the body, recurrences from the original primary cancer after remission, and second primary cancers (this is a new primary cancer in a person with a history of previous cancer that is different from the new primary cancer).

[0130] Stereoisomers described herein may include cis and trans isomers, optical isomers such as (R) and (S) enantiomers, diastereomers, geometric isomers, rotational isomers, atropisomers, conformational isomers and tautomers of the compounds of the invention (including compounds exhibiting more than one isomeric type); and mixtures thereof (e.g., racemates and diastereomeric pairs).

[0131] The compounds of the present invention can exhibit tautomerism and structural isomerism. For example, the compound can exist in several tautomeric forms, including enol and imine forms and ketone and enamine forms, and geometric isomers and mixtures thereof. All such tautomeric forms are included in the scope of the compounds of the present invention. Tautomers exist as mixtures of tautomeric groups in solution. In solid forms, usually one tautomer is dominant. Even if one tautomer can be described, the present invention includes all tautomers of the compounds provided.

[0132] Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC) or supercritical fluid chromatography (SFC).

[0133] The enantiomeric purity of the compounds described herein can be described in terms of enantiomeric excess (ee), which indicates the extent to which a sample contains one enantiomer in greater amounts than the other enantiomer. A racemic mixture has an ee of 0%, while a single, completely pure enantiomer has an ee of 100%. Similarly, diastereomeric purity can be described in terms of diastereomeric excess (de).

[0134] Synthesis method of the compound of the present invention

[0135] The present invention adopts the following scheme to prepare the compound represented by the general formula (I) of the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof.

[0136]

[0137] Step 1: In a suitable solvent and in the presence of a suitable base, compound Ia undergoes a substitution reaction with isopropyl pinacol borate to obtain compound Ib; the solvent is preferably tetrahydrofuran, the base is preferably lithium diisopropylamide, and the reaction temperature is between low temperature (such as -78°C) and room temperature;

[0138] Step 2: In a suitable solvent, in the presence of a suitable catalyst, under inert gas protection, compound Ic reacts with compound Ib to obtain compound Id; the solvent is preferably 1,4-dioxane, and the catalyst is preferably Pd(dppf)Cl 2 The inert gas is preferably nitrogen, and the reaction temperature is between room temperature and 100°C;

[0139] Step 3: In a suitable solvent, in the presence of a suitable base, compound Id reacts with compound Ie to obtain a compound of formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof; the solvent is, for example, N-methylpyrrolidone, the base is, for example, N,N-diisopropylethylamine, and the reaction temperature can be between 100° C. and 140° C.;

[0140] Among them, rings A, X, Y, Z, Q, R 3 , R 4 , R 5 , R 6 and q are as defined in the general formula (I). DETAILED DESCRIPTION

[0141] The compounds of the present invention and their preparation are further understood by way of examples, which illustrate some methods of preparing or using the compounds. However, it is to be understood that these examples do not limit the scope of the present invention. Variations of the present invention as now known or further developed are considered to fall within the scope of the present invention as described herein and as claimed.

[0142] The compounds of the present invention are prepared using convenient starting materials and common preparation steps. The present invention provides typical or preferred reaction conditions, such as reaction temperature, time, solvent, pressure, and molar ratio of reactants. However, unless otherwise specified, other reaction conditions can also be adopted. The optimized conditions may change with the use of specific reactants or solvents, but in general, the reaction optimized steps and conditions can be determined.

[0143] In addition, some protecting groups may be used in the present invention to protect certain functional groups from unnecessary reactions. Protecting groups suitable for various functional groups and their protection or deprotection conditions are widely known to those skilled in the art. For example, TW Greene and GM Wuts's "Protective Groups in Organic Preparations" (3rd edition, Wiley, New York, 1999 and references therein) describes in detail the protection or deprotection of a large number of protecting groups.

[0144] The separation and purification of compounds and intermediates can be carried out by appropriate methods and steps according to specific needs, such as filtration, extraction, distillation, crystallization, column chromatography, preparative thin layer plate chromatography, preparative high performance liquid chromatography or a combination of the above methods. The specific use method can refer to the examples described in the present invention. Of course, other similar separation and purification means can also be adopted. Conventional methods (including physical constants and spectral data) can be used to characterize them.

[0145] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The NMR measurements were performed using a Bruker 300 NMR spectrometer and the solvent was deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated chloroform (CDCl 3 ), deuterated methanol (CD 3 OD), and the internal standard was tetramethylsilane (TMS).

[0146] MS was measured using LC (Waters 2695) / MS (Quattro Premier xE) mass spectrometer (manufacturer: Waters) (Photodiode Array Detector).

[0147] The preparative liquid chromatography method used an LC6000 high performance liquid chromatograph (manufacturer: Chuangxin Tongheng), the chromatographic column was Daisogel C18 10 μm 100A (30 mm×250 mm), and the mobile phase was acetonitrile / water.

[0148] The thin layer chromatography silica gel plate used was Qingdao Ocean Chemical GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) had a specification of 0.20 mm to 0.25 mm, and the specification used in preparing the thin layer chromatography separation and purification product was 0.5 mm.

[0149] Column chromatography generally uses Qingdao marine silica gel 100-200 mesh, 200-300 mesh and 300-400 mesh silica gel as the carrier.

[0150] The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from online shopping malls, Beijing Coupling, Sigma, Bailingwei, Yishiming, Shanghai Shuya, Shanghai Inokai, Anaiji Chemical, Shanghai Bid, etc.

[0151] Unless otherwise specified in the examples, all reactions were carried out under a nitrogen atmosphere.

[0152] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1L.

[0153] The reaction solvent, organic solvent or inert solvent is expressed as the solvent used that does not participate in the reaction under the described reaction conditions, including benzene, toluene, acetonitrile, tetrahydrofuran (THF), dimethylformamide (DMF), chloroform, dichloromethane, ether, methanol, nitrogen-methylpyrrolidone (NMP), pyridine, etc. Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0154] The chemical reactions described in the present invention are generally carried out under normal pressure. The reaction temperature is between -78°C and 200°C. The reaction time and conditions are, for example, between -78°C and 200°C at one atmosphere, and are completed within about 1 to 24 hours. If the reaction is left overnight, the reaction time is generally 16 hours. In the embodiments, unless otherwise specified, the reaction temperature is room temperature, which is 20°C to 30°C.

[0155] The reaction progress in the examples was monitored by thin layer chromatography (TLC), and the developing solvent systems used in the reaction were: A: dichloromethane and methanol system, B: petroleum ether and ethyl acetate system, C: acetone, and the volume ratio of the solvent was adjusted according to the polarity of the compound.

[0156] The eluent system of column chromatography and the developing solvent system of thin layer chromatography used for purifying compounds include: A: dichloromethane and methanol system, B: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and trifluoroacetic acid can also be added for adjustment.

[0157] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any method and material similar or equivalent to the described content can be applied to the method of the present invention.

[0158] Example

[0159] Example 1: Preparation of (3S,4R)-4-((4-(3-cyclopentyl-2,6-dimethyl-3H-thieno[2,3-d]imidazol-5-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (1)

[0160]

[0161] Step 1: Preparation of methyl 2-(cyclopentylamino)-4-methylthiophene-3-carboxylate (1a)

[0162] At room temperature, methyl 2-amino-4-methylthiophene-3-carboxylate (5.00 g, 29.23 mmol), cyclopentanone (6.14 g, 73.08 mmol), glacial acetic acid (4.39 g, 73.08 mmol) and DCE (6.13 g, 44.37 mmol) were added to the reaction bottle, sodium triacetyl borohydride (15.49 g, 73.08 mmol) was slowly added, and the reaction was stirred at room temperature overnight. After the reaction was completed, a saturated sodium bicarbonate aqueous solution was added to quench the reaction, dichloromethane (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 5.53 g of the title product as a red oily liquid, with a yield of 79.1%.

[0163] LCMS: m / z 240.1 [M+H] + .

[0164] Step 2: Preparation of 2-(cyclopentylamino)-4-methylthiophene-3-carboxylic acid (1b)

[0165] At room temperature, methyl 2-(cyclopentylamino)-4-methylthiophene-3-carboxylate (3.00 g, 12.54 mmol), sodium bicarbonate (5.02 g, 125.4 mmol), water (15 mL) and anhydrous methanol (15 mL) were added to the reaction flask, heated to 80°C and stirred to react overnight, and the reaction was completed. After the reaction was completed, 2N HCl was added to adjust the pH to 3-4, ethyl acetate (50 mL) and water (20 mL) were added, the aqueous phase was extracted with ethyl acetate (50 mL*2), the organic phases were combined, dried and concentrated to obtain 2.70 g of the title product as a black oily liquid, with a yield of 95.2%.

[0166] LCMS: m / z 226.1[M+H] + .

[0167] Step 3: Preparation of 2-(N-cyclopentylacetylamino)-4-methylthiophene-3-carboxylic acid (1c)

[0168] At room temperature, 2-(cyclopentylamino)-4-methylthiophene-3-carboxylic acid (1.40 g, 5.86 mmol), acetic anhydride (1.45 g, 14.46 mmol), triethylamine (1.78 g, 17.58 mmol), DMAP (72.1 mg, 0.59 mmol) and dichloromethane (15 mL) were added to the reaction flask and stirred at room temperature overnight. After the reaction was completed, saturated sodium bicarbonate solution was added to quench the reaction, dichloromethane (50 mL) and water (20 mL) were added to take the organic phase, the aqueous phase was extracted with dichloromethane (30 mL*2), the organic phase was dried and concentrated, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 1.12 g of the title product as a red oily liquid with a yield of 71.3%.

[0169] LCMS: m / z 268.1[M+H] + .

[0170] Step 4: Preparation of N-(3-amino-4-methylthiophen-2-yl)-N-cyclopentylacetamide (1d)

[0171] At room temperature, 2-(N-cyclopentylacetylamino)-4-methylthiophene-3-carboxylic acid (1.12 g, 4.19 mmol), DPPA (2.88 g, 10.48 mmol), triethylamine (2.12 g, 20.95 mmol) and 1,4-dioxane (10 mL) were placed in a reaction bottle and stirred at room temperature for 1 h. Then, water (10 mL) was added to the reaction solution and the temperature was raised to 100 °C and stirred overnight. After the reaction was completed, 2N NaOH (10 mL) was added and stirred for 30 min, dichloromethane (50 mL) and water (50 mL) were added, and the organic phase was taken. The organic phase was washed with water (30 mL*3), the organic phase was taken and dried and concentrated, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 742 mg of the title product as a brown solid with a yield of 74.4%.

[0172] LCMS: m / z 239.1[M+H] + .

[0173] Step 5: Preparation of 3-cyclopentyl-2,6-dimethyl-3H-thieno[2,3-d]imidazole (1e).

[0174] At room temperature, N-(3-amino-4-methylthiophene-2-yl)-N-cyclopentylacetamide (554 mg, 2.33 mmol), phosphorus oxychloride (374.1 mg, 2.44 mmol) and toluene (10 mL) were placed in a reaction bottle, heated to 100°C and stirred to react overnight. After the reaction was completed, saturated sodium bicarbonate aqueous solution (10 mL) was added to quench the reaction, ethyl acetate (50 mL) and water (50 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was dried and concentrated, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 514 mg of the title product as a red solid, with a yield of 98.0%.

[0175] LCMS: m / z 221.1 [M+H] + .

[0176] Step 6: Preparation of 3-cyclopentyl-2,6-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3H-thieno[2,3-d]imidazole (1f).

[0177] At room temperature, 3-cyclopentyl-2,6-dimethyl-3H-thieno[2,3-d]imidazole (740 mg, 3.36 mmol), isopropyl pinacol borate (1.25 g, 6.72 mmol) and THF (10 mL) were placed in a reaction bottle, nitrogen was replaced, the temperature was lowered to -78°C, LDA (8.40 mL, 16.8 mmol) was slowly added, the reaction was stirred overnight and slowly warmed to room temperature. After the reaction was completed, saturated aqueous ammonium chloride solution (10 mL) was added to quench the reaction, ethyl acetate (50 mL) and water (50 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was taken and dried and concentrated to obtain 1.16 g of the title product as a red solid, which was directly used in the next step.

[0178] LCMS: m / z 347.2 [M+H] + .

[0179] Step 7: Preparation of 5-(2-chloro-5-fluoropyrimidin-4-yl)-3-cyclopentyl-2,6-dimethyl-3H-thieno[2,3-d]imidazole (1 g).

[0180] At room temperature, 3-cyclopentyl-2,6-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3H-thieno[2,3-d]imidazole (1.16 g, crude), 2,4-dichloro-5-fluoropyrimidine (836.64 mg, 5.04 mmol), Pd(dppf)Cl 2(248.54 mg, 0.34 mmol), potassium carbonate (1.39 g, 10.08 mmol), 1,4-dioxane (10 mL) and water (3 mL) were added to the reaction flask, the nitrogen was replaced, the temperature was raised to 100 ° C, and the reaction was stirred overnight. After the reaction was completed, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was dried and concentrated, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 415 mg of the title product as a brown solid with a yield of 35.2%.

[0181] LCMS: m / z 351.1 [M+H] + .

[0182] Step 8: Preparation of (3S,4R)-4-((4-(3-cyclopentyl-2,6-dimethyl-3H-thieno[2,3-d]imidazol-5-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (1).

[0183] At room temperature, 5-(2-chloro-5-fluoropyrimidin-4-yl)-3-cyclopentyl-2,6-dimethyl-3H-thieno[2,3-d]imidazole (100 mg, 0.29 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (44.4 mg, 0.29 mmol), DIEA (74.96 mg, 0.58 mmol) and N-methylpyrrolidone (2 mL) were added to the reaction bottle and stirred at 120°C overnight. After cooling to room temperature, ethyl acetate (20 mL) and water (5 mL) were added. The organic phase was dried and concentrated. The residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 60 mg of the title compound as a brown solid, with a yield of 47.9%.

[0184] LCMS: m / z 432.2 [M+H] + .

[0185] 1H NMR (400MHz, DMSO) δ8.34(t,J=7.9Hz,1H),7.05(d,J=7.7Hz,1H),4.90(d,J=5.3Hz ,1H),4.88–4.71(m,1H),3.90–3.68(m,3H),3.50(ddd,J=14.3,9.5,4.9Hz,1H),3.1 1–3.00(m,1H),2.58–2.53(m,3H),2.28–2.08(m,3H),1.99(d,J=12.2Hz,1H),1.86( dd,J=11.8,7.6Hz,5H), 1.71(d,J=3.4Hz,3H), 1.46(ddd,J=15.8,11.9,4.4Hz,1H).

[0186] Example 2: Preparation of (3S,4R)-4-((5-fluoro-4-(7-isopropylthieno[3,2-b]pyridin-2-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (2)

[0187]

[0188] Step 1: Preparation of 7-(prop-1-en-2-yl)thieno[3,2-b]pyridine (2a)

[0189] At room temperature, 7-chlorothieno[3,2-b]pyridine (2.50 g, 14.79 mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (3.45 g, 17.75 mmol), Pd(dppf)Cl2 (1.07 g, 1.47 mmol), potassium carbonate (6.13 g, 44.37 mmol), 1,4-dioxane (25 mL) and water (5 mL) were added to a reaction flask, the atmosphere was replaced with nitrogen, the temperature was raised to 100°C and the reaction was stirred overnight. After the reaction, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, which was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 3.44 g of the title product as a yellow oily liquid with a yield of 80.1%.

[0190] LCMS: m / z 176.0 [M+H] + .

[0191] Step 2: Preparation of 7-isopropylthieno[3,2-b]pyridine (2b)

[0192] At room temperature, 7-(prop-1-en-2-yl)thieno[3,2-b]pyridine (500 mg, 2.86 mmol), palladium carbon (100 mg) and anhydrous methanol (10 mL) were added to the reaction flask, hydrogen was replaced, and the reaction was stirred at room temperature overnight to complete the reaction. After the reaction was completed, diatomaceous earth was used for filtration, the filtrate was concentrated, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 580 mg of the title product as a colorless oily liquid, with a yield of 96.6%.

[0193] LCMS: m / z 178.1[M+H] + .

[0194] Step 3: Preparation of (7-isopropylthieno[3,2-b]pyridin-2-yl)boronic acid (2c)

[0195] At room temperature, 7-isopropylthieno[3,2-b]pyridine (300 mg, 1.69 mmol) and THF (5 mL) were added to a three-necked flask, nitrogen was replaced, the temperature was lowered to -78°C, n-butyl lithium (0.81 mL, 2.03 mmol) was added, and the reaction was stirred for 1 hour. Then 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborane (628.9 mg, 3.38 mmol) was added to the reaction solution, and the reaction was stirred overnight, and the temperature was slowly raised to room temperature. After the reaction was completed, saturated ammonium chloride solution was added to quench the reaction, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was taken and dried and concentrated to obtain 577 mg of a crude white solid product, which was directly used in the next step.

[0196] LCMS: m / z 222.1[M+H] + .

[0197] Step 4: Preparation of 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-isopropylthieno[3,2-b]pyridine (2d)

[0198] At room temperature, (7-isopropylthieno[3,2-b]pyridin-2-yl)boronic acid (577 mg, 2.61 mmol), 2,4-dichloro-5-fluoropyrimidine (650.7 mg, 3.92 mmol), Pd(dppf)Cl 2(190.0 mg, 0.26 mmol), potassium carbonate (1.08 g, 1.86 mmol), 1,4-dioxane (10 mL) and water (3 mL) were added to the reaction flask, the nitrogen was replaced, and the reaction was stirred at 100°C overnight. After the reaction was completed, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was dried and concentrated, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 170 mg of the title product as a light yellow solid, with a yield of 21.2%.

[0199] LCMS: m / z 308.0 [M+H] + .

[0200] Step 5: Preparation of (3S,4R)-4-((5-fluoro-4-(7-isopropylthieno[3,2-b]pyridin-2-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (2).

[0201] At room temperature, 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-isopropylthieno[3,2-b]pyridine (100 mg, 0.33 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (50.5 mg, 0.33 mmol), DIEA (85.3 mg, 0.66 mmol) and NMP (2 mL) were added to the reaction flask and stirred at 120°C overnight. After cooling to room temperature, ethyl acetate (20 mL) and water (5 mL) were added. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. The residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 31 mg of the title compound as a light yellow solid, with a yield of 24.2%.

[0202] LCMS: m / z 389.1[M+H] + .

[0203] 1H NMR (400MHz, DMSO) δ8.68(t,J=6.6Hz,1H),8.54(d,J=3.2Hz,1H),8.18(d,J=1.6Hz,1H),7.42–7.25(m,2H),4.94(d,J=5.1Hz,1H),3.94–3.75(m ,3H),3.53(dd,J=8.8,4.5Hz,1H),3.41–3.35(m,1H),3.23(dt,J=13.6,6.9Hz,1H),3.08(t,J=10.2Hz,1H),1.99(s,1H),1.39(t,J=8.6Hz,6H).

[0204] Example 3: Preparation of (3S,4R)-4-((4-(7-cyclopentylthieno[3,2-b]pyridin-2-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (3)

[0205]

[0206]

[0207] Step 1: Preparation of 7-(cyclopent-1-en-1-yl)thieno[3,2-b]pyridine (3a)

[0208] At room temperature, 7-chlorothieno[3,2-b]pyridine (5.00 g, 29.59 mmol), 2-(cyclopent-1-en-1-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (6.89 g, 35.51 mmol), Pd(dppf)Cl 2 (2.16g, 2.96mmol), potassium carbonate (12.40g, 89.85mmol), 1,4-dioxane (50mL), and water (12mL) were added to the reaction flask, the nitrogen was replaced, and the temperature was raised to 100°C and stirred to react overnight. After the reaction was completed, ethyl acetate (50mL) and water (20mL) were added to take the organic phase, which was washed with water (30mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 5.50g of the title product as a yellow oily liquid with a yield of 92.4%.

[0209] LCMS: m / z 202.1[M+H] + .

[0210] Step 2: Preparation of 7-cyclopentylthieno[3,2-b]pyridine (3b)

[0211] At room temperature, 7-(cyclopent-1-en-1-yl)thieno[3,2-b]pyridine (5.50 g, 27.21 mmol), palladium carbon (800 mg) and anhydrous methanol (10 mL) were added to the reaction flask, hydrogen was replaced, and the reaction was stirred at room temperature overnight to complete the reaction. After the reaction was completed, diatomaceous earth was used for filtration, the filtrate was concentrated, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 6.29 g of the title product as a colorless oily liquid, with a yield of 96.6%.

[0212] LCMS: m / z 204.1[M+H] + .

[0213] Step 3: Preparation of (7-cyclopentylthieno[3,2-b]pyridin-2-yl)boronic acid (3c)

[0214] At room temperature, 7-cyclopentylthieno[3,2-b]pyridine (2.00 g, 9.85 mmol) and THF (20 mL) were added to a three-necked flask, nitrogen was replaced, the temperature was lowered to -78°C, n-butyl lithium (4.73 mL, 11.82 mmol) was added, and the reaction was stirred for 1 h. Then 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborane (3.67 g, 19.7 mmol) was added to the reaction solution and stirred for overnight, while slowly warming to room temperature. After the reaction was completed, saturated ammonium chloride solution was added to quench the reaction, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was taken and dried and concentrated to obtain 2.43 g of a crude brown oily liquid product, which was directly used in the next step.

[0215] LCMS: m / z 248.1[M+H] + .

[0216] Step 4: Preparation of 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-cyclopentylthieno[3,2-b]pyridine (3d)

[0217] At room temperature, (7-cyclopentylthieno[3,2-b]pyridin-2-yl)boronic acid (2.43 g, crude), 2,4-dichloro-5-fluoropyrimidine (2.45 g, 14.78 mmol), Pd(dppf)Cl 2 (723.6 mg, 0.99 mmol), potassium carbonate (4.10 g, 29.55 mmol), 1,4-dioxane (30 mL) and water (10 mL) were added to the reaction flask, the nitrogen was replaced, and the reaction was stirred at 100°C overnight. After the reaction was completed, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was dried and concentrated, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 2.0 g of the title product as a yellow solid with a yield of 60.8%.

[0218] LCMS: m / z 334.1[M+H] + .

[0219] Step 5: Preparation of (3S,4R)-4-((4-(7-cyclopentylthieno[3,2-b]pyridin-2-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (3).

[0220] At room temperature, 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-cyclopentylthieno[3,2-b]pyridine (100 mg, 0.30 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (45.9 mg, 0.30 mmol), DIEA (77.54 mg, 0.60 mmol) and N-methylpyrrolidone (NMP) (2 mL) were added to the reaction bottle and stirred at 120°C overnight. After cooling to room temperature, ethyl acetate (20 mL) and water (5 mL) were added. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. The residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 31 mg of the title compound as a light yellow solid, with a yield of 24.2%.

[0221] LCMS: m / z 415.2 [M+H] + .

[0222] 1H NMR (400MHz, CDCl3) δ8.67(dd,J=18.8,4.8Hz,1H),8.33(d,J=1.6Hz,1H),8.20(t,J=24.1Hz,1H),7.19(d,J=4.8Hz,1H ),5.25(d,J=6.4Hz,1H),4.09(dt,J=24.4,12.2Hz,1H),4.00(dd,J=11.6,3.7Hz,1H),3.89(dt,J=11.2,8.8Hz,1H),3. 66(td,J=9.4,4.9Hz,1H),3.52(td,J=11.7,2.1Hz,1H),3.36(dd,J=16.0,8.1Hz,1H),3.25(dd,J=21.3,11.3Hz,1H),2 .26(dd,J=9.9,7.1Hz,2H),2.17–2.03(m,1H),1.83(ddd,J=10.3,6.0,3.7Hz,4H),1.71(ddd,J=24.6,11.8,4.8Hz,2H).

[0223] Example 4: Preparation of (3S,4R)-4-((4-(3-chloro-7-cyclopentylthieno[3,2-b]pyridin-2-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (4)

[0224]

[0225]

[0226] Step 1: Preparation of 3-chloro-7-cyclopentylthieno[3,2-b]pyridine (4a)

[0227] At room temperature, 7-cyclopentylthieno[3,2-b]pyridine (2.00 g, 9.85 mmol), N-chlorosuccinimide (1.60 g, 11.82 mmol) and trifluoromethanesulfonic acid (5 mL) were added to the reaction flask, and the reaction was stirred at room temperature overnight. After the reaction was completed, a saturated sodium bicarbonate aqueous solution was added to adjust the pH to 7-8, and ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, which was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 1.72 g of the title product as a yellow oily liquid with a yield of 73.7%.

[0228] LCMS: m / z 238.0 [M+H] + .

[0229] Step 2: Preparation of (3-chloro-7-cyclopentylthieno[3,2-b]pyridin-2-yl)boronic acid (4b)

[0230] At room temperature, 3-chloro-7-cyclopentylthieno[3,2-b]pyridine (500 mg, 2.11 mmol) and THF (20 mL) were added to a three-necked flask, nitrogen was replaced, the temperature was lowered to -78°C, n-butyl lithium (3.38 mL, 8.44 mmol) was added, and the reaction was stirred for 1 hour. Then 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborane (980.5 mg, 5.27 mmol) was added to the reaction solution, the reaction was stirred overnight, and the temperature was slowly raised to room temperature. After the reaction was completed, saturated ammonium chloride solution was added to quench the reaction, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was taken and dried and concentrated to obtain 600 mg of a crude brown oily liquid product, which was directly used in the next step.

[0231] LCMS: m / z 282.0 [M+H] + .

[0232] Step 3: Preparation of 3-chloro-2-(2-chloro-5-fluoropyrimidin-4-yl)-7-cyclopentylthieno[3,2-b]pyridine (4c)

[0233] At room temperature, (3-chloro-7-cyclopentylthieno[3,2-b]pyridin-2-yl)boronic acid (600 mg, crude), 2,4-dichloro-5-fluoropyrimidine (526.2 mg, 3.17 mmol), Pd(dppf)Cl 2(153.5 mg, 0.21 mmol), potassium carbonate (874.87 mg, 6.33 mmol), 1,4-dioxane (10 mL) and water (3 mL) were added to the reaction flask, the nitrogen was replaced, and the reaction was stirred at 100°C overnight. After the reaction was completed, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was dried and concentrated, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 140 mg of the title product as a red oily liquid with a yield of 18.1%.

[0234] LCMS: m / z 368.0 [M+H] + .

[0235] Step 4: Preparation of (3S,4R)-4-((4-(3-chloro-7-cyclopentylthieno[3,2-b]pyridin-2-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (4)

[0236] At room temperature, 3-chloro-2-(2-chloro-5-fluoropyrimidin-4-yl)-7-cyclopentylthieno[3,2-b]pyridine (110 mg, 0.38 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (58.14 mg, 0.38 mmol), DIEA (98.2 mg, 0.76 mmol) and N-methylpyrrolidone (NMP) (2 mL) were added to the reaction bottle and stirred at 120°C overnight. After cooling to room temperature, ethyl acetate (20 mL) and water (5 mL) were added. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. The residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 60 mg of the title compound as a light yellow solid, with a yield of 35.2%.

[0237] LCMS: m / z 449.1[M+H] + .

[0238] 1H NMR (400MHz, CDCl3) δ8.78(s,1H),8.34(s,1H),7.29(s,1H),5.35(d,J=4.4Hz,1H),4.08(dd,J=11.2,4.6Hz,1H),3.99(dd,J=11.2,2.7Hz,1H),3.95 –3.83(m,1H),3.65(td,J=9.3,4.9Hz,1H),3.48(t,J=11.2Hz,1H),3.43–3 .30(m,1H),3.22(t,J=10.5Hz,1H),2.34–2.05(m,4H),2.05–1.60(m,7H).

[0239] Example 5: Preparation of 2-(5-fluoro-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-4-isopropylthieno[3,2-b]pyridine 7-oxide (5)

[0240]

[0241] Step 1: Preparation of 2-(2-chloro-5-fluoropyrimidin-4-yl)-4-isopropylthieno[3,2-b]pyridine 7-oxide (5a)

[0242] At room temperature, 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-isopropylthieno[3,2-b]pyridine (400 mg, 1.30 mmol), mCPBA (376.2 mg, 2.18 mmol) and DCM (10 mL) were added to the reaction flask and stirred at room temperature for 2 h. After the reaction was completed, a saturated sodium thiosulfate solution was added to quench the reaction, and dichloromethane (50 mL) and water (20 mL) were added to take the organic phase, which was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure. The crude product was purified by column chromatography (eluent: dichloromethane: methanol = 10:1) to obtain 210 mg of the title product as a yellow solid with a yield of 50.0%.

[0243] LCMS: m / z 324.0 [M+H] + .

[0244] Step 2: Preparation of 2-(5-fluoro-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-4-isopropylthieno[3,2-b]pyridine 7-oxide (5)

[0245] At room temperature, 2-(2-chloro-5-fluoropyrimidin-4-yl)-4-isopropylthieno[3,2-b]pyridine 7-oxide (100 mg, 0.31 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (47.43 mg, 0.31 mmol), cesium carbonate (202.12 mg, 0.62 mmol), palladium acetate (6.98 mg, 0.031 mmol), BINAP (38.56 mg, 0.062 mmol) and 1,4-dioxane (5 mL) were added to the reaction flask, the atmosphere was replaced with nitrogen, the temperature was raised to 100°C, and the reaction was stirred overnight. After the reaction, the filtrate was collected by diatomaceous earth filtration, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was dried and concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 16 mg of the light yellow solid title compound, with a yield of 12.8%.

[0246] LCMS: m / z 405.1[M+H] + .

[0247] 1H NMR (400MHz, DMSO) δ8.57(d,J=3.1Hz,1H),8.37(d,J=6.5Hz,1H),8.26(d,J=1.4Hz,1H),7.39(t,J=7.0Hz,2H),4.94(d,J=5.3Hz,1H),3.84(dd,J= 10.9,5.0Hz,3H),3.59–3.47(m,1H),3.38(d,J=11.5Hz,1H),3.22–3.14( m,2H),3.07(t,J=10.2Hz,1H),1.56–1.45(m,1H),1.37(d,J=6.8Hz,6H).

[0248] Example 6: Preparation of 2-(5-fluoro-2-((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-7-isopropyl-3,5-dimethylthieno[3,2-c]pyridin-4(5H)-one (6)

[0249]

[0250] Step 1: Preparation of 1-(5-bromo-4-methylthiophen-2-yl)-2-methylpropan-1-one (6a).

[0251] At 0°C, aluminum chloride (4.12 g, 31.3 mmol) was dissolved in dichloromethane (50 mL), and isobutyryl chloride (3.31 g, 31.3 mmol) was slowly added dropwise. After stirring for 10 min, 2-bromo-3-thiophene (5.00 g, 28.4 mmol) was added dropwise. The reaction solution was slowly warmed to room temperature and stirred for 16 h. After monitoring the reaction completion, the reaction solution was poured into ice water (50 mL) and stirred for 30 min. The mixture was allowed to stand for stratification. The aqueous phase was extracted with dichloromethane (30 mL x 2). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain 6.9 g of the title compound as a yellow solid with a yield of 98%.

[0252] LCMS: m / z 246.9 [M+H] + .

[0253] Step 2: Preparation of ethyl 3-(5-bromo-4-methylthiophen-2-yl)-4-methylpent-2-enoate (6b).

[0254] At 0°C, sodium hydride (1.44 g, 36.2 mmol) (60% dissolved in mineral oil) was added to tetrahydrofuran (35 mL), and ethyl 2-(diethoxyphosphoryl)acetate (8.10 g, 36.2 mmol) was slowly added dropwise. After stirring for 20 min, a solution of 1-(5-bromo-4-methylthiophen-2-yl)-2-methylpropan-1-one (6a) (4.45 g, 18.1 mmol) in tetrahydrofuran (10 mL) was added. The reaction solution was stirred at room temperature for 60 h. After the reaction was completed, the reaction solution was poured into ice water (40 mL), ethyl acetate (30 mL) was added, and the stratification was allowed to stand. The aqueous phase was extracted with ethyl acetate (30 mL x 2), and the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain 5.2 g of the title compound as a yellow liquid with a yield of 89%.

[0255] LCMS: m / z 317.00 [M+H] + .

[0256] Step 3: Preparation of 3-(5-bromo-4-methylthiophen-2-yl)-4-methylpent-2-enoic acid (6c).

[0257] At room temperature, ethyl 3-(5-bromo-4-methylthiophen-2-yl)-4-methylpent-2-enoate (6b) (3.90 g, 12.3 mmol) was dissolved in a mixed solvent of methanol (40 mL) and water (10 mL), and lithium hydroxide monohydrate (2.07 g, 49.2 mmol) was added. The reaction solution was stirred at 50°C for 16 h. After monitoring the completion of the reaction, the reaction solution was concentrated to 10 mL, adjusted to pH 5-6 with 2M dilute hydrochloric acid, extracted with ethyl acetate (20 mL x 3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain 3.1 g of the title compound as a yellow solid with a yield of 88%. The residue was used directly in the next step without further purification.

[0258] LCMS: m / z 288.90 [M+H] + .

[0259] Step 4: Preparation of 2-bromo-7-isopropyl-3-methylthieno[3,2-c]pyridin-4(5H)-one (6d).

[0260] At room temperature, 3-(5-bromo-4-methylthiophen-2-yl)-4-methylpent-2-enoic acid (6c) (6.6 g, 22.9 mmol) and triethylamine (6.98 g, 68.7 mmol) were dissolved in diphenyl ether (50 mL), and diphenylphosphoryl azide (9.45 g, 34.3 mmol) was added dropwise, and the reaction solution was stirred at 150°C for 30 min. After monitoring the completion of the reaction, the reaction solution was concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 100% petroleum ether ~ 2: 1) to obtain 2.9 g of the title compound as a yellow solid, with a yield of 44%.

[0261] LCMS: m / z 285.90 [M+H] + .

[0262] Step 5: Preparation of 2-bromo-7-isopropyl-3,5-dimethylthieno[3,2-c]pyridin-4(5H)-one (6e).

[0263] At room temperature, 2-bromo-7-isopropyl-3-methylthieno[3,2-c]pyridin-4(5H)-one (6d) (500 mg, 1.75 mmol) and cesium carbonate (1.14 g, 3.51 mmol) were dissolved in N,N-dimethylformamide (5 mL), iodomethane (498 mg, 3.51 mmol) was added, and the reaction solution was stirred at room temperature for 1 h. After monitoring the completion of the reaction, the reaction solution was concentrated, diluted with ethyl acetate (20 mL), washed with water (10 mL x 3), and then washed with saturated brine (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain 465 mg of the title compound as a light yellow solid, with a yield of 89%.

[0264] LCMS: m / z 299.90 [M+H] + .

[0265] Step 6: Preparation of 7-isopropyl-3,5-dimethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thieno[3,2-c]pyridin-4(5H)-one (6f).

[0266] At room temperature, 2-bromo-7-isopropyl-3,5-dimethylthieno[3,2-c]pyridin-4(5H)-one (6e) (460 mg, 1.54 mmol), bis(boronic acid pinacol) (586 mg, 2.31 mmol) and potassium acetate (302 mg, 3.08 mmol) were dissolved in 1,4-dioxane (5 mL), and Pd(dppf)Cl was added. 2 (113 mg, 0.154 mmol), the reaction solution was replaced with gas three times under nitrogen protection, and stirred at 100 ° C for 2 h. After monitoring the reaction completion, ethyl acetate (15 mL) was added to dilute the reaction solution, filtered and concentrated to dryness to obtain 530 mg of the crude title compound as a black solid. The crude product was used directly in the next step without further purification.

[0267] LCMS: m / z 348.00 [M+H] + .

[0268] Step 7: Preparation of 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-isopropyl-3,5-dimethylthieno[3,2-c]pyridin-4(5H)-one (6 g).

[0269] 7-Isopropyl-3,5-dimethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thieno[3,2-c]pyridin-4(5H)-one (6f) (530 mg, 1.53 mmol), potassium phosphate (653 mg, 3.08 mmol) and 2,4-dichloro-5-fluoropyrimidine (386 mg, 2.31 mmol) were dissolved in a mixed solvent of 1,4-dioxane (6 mL) and water (1.5 mL) at room temperature, and Pd(dppf)Cl was added. 2 (113 mg, 0.154 mmol), the reaction solution was replaced with gas three times under nitrogen protection, and stirred at 100 ° C for 16 h. After monitoring the reaction completion, the reaction solution was cooled to room temperature, ethyl acetate (20 mL) and water (15 mL) were added, and the layers were separated by standing. The aqueous phase was extracted with ethyl acetate (20 mL x 2), and the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain 180 mg of the title compound as a yellow oil, with a two-step yield of 33%.

[0270] LCMS: m / z 351.90 [M+H] + .

[0271] Step 8: Preparation of 2-(5-fluoro-2-((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-7-isopropyl-3,5-dimethylthieno[3,2-c]pyridin-4(5H)-one (6).

[0272] At room temperature, 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-isopropyl-3,5-dimethylthieno[3,2-c]pyridin-4(5H)-one (6 g) (150 mg, 0.427 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (65.3 mg, 0.427 mmol) and diisopropylethylamine (110 mg, 0.855 mmol) were dissolved in N-methylpyrrolidone (50 mL), and the reaction solution was stirred at 120 °C for 16 h. After monitoring the completion of the reaction, the reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 mL x 3), washed with saturated brine (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to give 45.0 mg of the title compound as a light yellow solid, with a yield of 24%.

[0273] LC-MS: m / z 433.00 [M+H] + .

[0274] 1 H NMR(400MHz,DMSO)δ8.45(d,J=2.8Hz,1H),7.51(s,1H),7.26(d,J=7.8Hz,1H), 4.90(d,J=5.3Hz,1H),3.82(dd,J=11.0,5.1Hz,3H),3.59-3.41(m,4H),3.34(s, 1H),3.08-2.98(m,1H),2.87(dt,J=13.7,6.8Hz,1H),2.66(d,J=2.6Hz,3H),1.9 6(d,J=11.7Hz,1H), 1.47(ddd,J=16.0,12.1,4.5Hz,1H), 1.30(d,J=6.9Hz,6H).

[0275] Example 7: Preparation of (3S,4R)-4-((5-fluoro-4-(3-isopropyl-2,6-dimethyl-3H-thieno[2,3-d]imidazol-5-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (7)

[0276]

[0277] Step 1: Preparation of ethyl 2-(isopropylamino)-4-methylthiophene-3-carboxylate (7a).

[0278] At room temperature, ethyl 2-amino-4-methylthiophene-3-carboxylate (5.00 g, 0.0270 mol), 2-methoxypropyl-1-ene (2.92 g, 0.0405 mmol) and glacial acetic acid (2.43 g, 0.0405 mol) were dissolved in 1,2-dichloroethane (50 mL), sodium triacetoxyborohydride (8.58 g, 0.0405 mol) was added, and the reaction solution was stirred at room temperature for 16 h. After monitoring the reaction completion, saturated sodium bicarbonate aqueous solution was added and stirred for 10 min, extracted with dichloromethane (50 mL x 3), and the combined organic phase was washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was directly used in the next step to obtain 6.10 g of the title compound as a brown solid, with a yield of 99%.

[0279] LCMS: m / z 228.00 [M+H] + .

[0280] Step 2: Preparation of 2-(isopropylamino)-4-methylthiophene-3-carboxylic acid (7b).

[0281] At room temperature, ethyl 2-(isopropylamino)-4-methylthiophene-3-carboxylate (7a) (6.10 g, 0.0268 mmol) and lithium hydroxide monohydrate (4.51 g, 0.107 mmol) were dissolved in a mixed solvent of ethanol (60 mL) and water (12 mL), and the reaction solution was stirred at 80°C for 16 h. After monitoring the completion of the reaction, the reaction solution was concentrated, and the pH was adjusted to 5-6 with 2N dilute hydrochloric acid. The aqueous phase was extracted with ethyl acetate (30 mL x 3), and the combined organic phase was washed with saturated brine (20 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was directly used in the next step to obtain 5.1 g of the title compound as a yellow solid with a yield of 95%.

[0282] LCMS: m / z 200.00 [M+H] + .

[0283] Step 3: Preparation of 2-(N-isopropylacetylamino)-4-methylthiophene-3-carboxylic acid (7c).

[0284] At room temperature, 2-(isopropylamino)-4-methylthiophene-3-carboxylic acid (7b) (5.40 g, 0.0271 mol), triethylamine (8.21 g, 0.0542 mol) and DMAP (0.331 g, 0.00271 mol) were dissolved in dichloromethane (60 mL), and acetic anhydride (5.53 g, 0.0542 mmol) was added dropwise. The reaction solution was stirred at room temperature for 16 h. After monitoring the reaction completion, water (40 mL) was added and the mixture was allowed to stand for stratification. The aqueous phase was extracted with dichloromethane (30 mL x 3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain 2.71 g of the title compound as a yellow solid, with a yield of 41%.

[0285] LCMS: m / z 242.00 [M+H] + .

[0286] Step 4: Preparation of N-(3-amino-4-methylthiophen-2-yl)-N-isopropylacetamide (7d).

[0287] At room temperature, 2-(N-isopropylacetylamino)-4-methylthiophene-3-carboxylic acid (7c) (2.50 g, 0.0104 mol) and triethylamine (3.15 g, 0.0312 mol) were dissolved in 1,4-dioxane (13 mL), and diphenylphosphoryl azide (4.29 g, 0.0156 mol) was added. The reaction solution was stirred at room temperature for 1 h, and then water (13 mL) was added and stirred at 100°C for 18 h. After monitoring the completion of the reaction, NaOH aqueous solution was added to adjust the pH to 8-9, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain 2.10 g of the title compound as a black oil with a yield of 95%.

[0288] LCMS: m / z 213.00 [M+H] + .

[0289] Step 5: Preparation of 3-isopropyl-2,6-dimethyl-3H-thieno[2,3-d]imidazole (7e).

[0290] At room temperature, N-(3-amino-4-methylthiophen-2-yl)-N-isopropylacetamide (7d) (2.10 g, 9.91 mmol) was dissolved in toluene (20 mL), phosphorus oxychloride (1.67 g, 10.9 mmol) was added, and the reaction solution was stirred at 100°C for 16 h. After monitoring the completion of the reaction, the reaction solution was concentrated, 10% sodium carbonate aqueous solution (20 mL) was added, and extracted with ethyl acetate (20 mL x 3), the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (dichloromethane: methanol = 10: 1) to obtain 1.30 g of the title compound as a brown solid, with a yield of 67%.

[0291] LCMS: m / z 195.00 [M+H] + .

[0292] Step 6: Preparation of 3-isopropyl-2,6-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3H-thieno[2,3-d]imidazole (7f).

[0293] 3-Isopropyl-2,6-dimethyl-3H-thieno[2,3-d]imidazole (7e) (500 mg, 2.57 mmol) and 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxolane (1.20 g, 6.44 mmol) were dissolved in tetrahydrofuran (10 mL) at -78°C, LDA (12.8 mL, 25.7 mmol) was added dropwise, and the reaction solution was stirred at -78°C for 3 h under nitrogen protection. Aqueous ammonium chloride solution was added to the reaction solution, and it was extracted with ethyl acetate (20 mL x 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain 650 mg of the title compound as a yellow oil, with a yield of 79%.

[0294] LCMS: m / z 321.00 [M+H] + .

[0295] Step 7: Preparation of 5-(2-chloro-5-fluoropyrimidin-4-yl)-3-isopropyl-2,6-dimethyl-3H-thieno[2,3-d]imidazole (7 g).

[0296] 3-Isopropyl-2,6-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3H-thieno[2,3-d]imidazole (7f) (650 mg, 2.03 mmol), potassium phosphate (860 mg, 4.06 mmol) and 2,4-dichloro-5-fluoropyrimidine (508 mg, 3.04 mmol) were dissolved in a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL) at room temperature, and Pd(dppf)Cl was added. 2 (148mg, 0.203mmol), the reaction solution was replaced with gas three times under nitrogen protection, and stirred at 100°C for 16h. After monitoring the reaction completion, ethyl acetate (20mL) was added to dilute the reaction solution, water (20mL) was added, and the layers were separated by standing. The aqueous phase was extracted with ethyl acetate (20mL x 3), and the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain 460mg of the title compound as a yellow oil, with a yield of 69%.

[0297] LCMS: m / z 325.00 [M+H] + .

[0298] Step 8: Preparation of (3S,4R)-4-((5-fluoro-4-(3-isopropyl-2,6-dimethyl-3H-thieno[2,3-d]imidazol-5-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (7).

[0299] 5-(2-Chloro-5-fluoropyrimidin-4-yl)-3-isopropyl-2,6-dimethyl-3H-thieno[2,3-d]imidazole (7 g) (250 mg, 0.772 mmol) was dissolved in N-methylpyrrolidone (3 mL) at room temperature, and diisopropylethylamine (199 mg, 1.54 mmol) and (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (118 mg, 0.772 mmol) were added. The reaction solution was stirred at 120 °C for 18 h. After monitoring the completion of the reaction, the reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 mL x 3), washed with saturated brine (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 87.0 mg of the title compound as a white solid, with a yield of 28%.

[0300] LC-MS: m / z 406.00 [M+H] + .

[0301] 1 H NMR (400MHz, DMSO) δ8.33(d,J=3.6Hz,1H),7.05(d,J=7.7Hz,1H),4.91(d,J=5.3Hz,1H),4.76-4.58(m,1H),3.91-3.68( m,3H),3.53-3.46(m,1H),3.37-3.26(m,4H),3.14-2.98(m,1H),2.51(s,3H),1.99(d,J=10.9Hz,1H),1.54-1.37(m,7H).

[0302] Example 8: Preparation of (3S,4R)-4-((5-fluoro-4-(3-isopropyl-2-methyl-3H-thieno[2,3-d]imidazol-5-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (8)

[0303]

[0304] Step 1: Preparation of ethyl 2-(isopropylamino)thiophene-3-carboxylate (8a).

[0305] At room temperature, ethyl 2-aminothiophene-3-carboxylate (5.00 g, 0.0290 mol), 2-methoxypropyl-1-ene (3.16 g, 0.0438 mmol) and glacial acetic acid (2.63 g, 0.0438 mol) were dissolved in 1,2-dichloroethane (50 mL), sodium triacetoxyborohydride (9.28 g, 0.0438 mol) was added, and the reaction solution was stirred at room temperature for 16 h. After monitoring the reaction completion, saturated sodium bicarbonate aqueous solution was added and stirred for 10 min, extracted with dichloromethane (50 mL x 3), and the combined organic phase was washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was directly used in the next step to obtain 6.10 g of the title compound as a brown solid with a yield of 98%.

[0306] LCMS: m / z 214.00 [M+H] + .

[0307] Step 2: Preparation of 2-(isopropylamino)thiophene-3-carboxylic acid (8b).

[0308] At room temperature, ethyl 2-(isopropylamino)thiophene-3-carboxylate (8a) (6.10 g, 0.0286 mmol) and lithium hydroxide monohydrate (4.80 g, 0.114 mmol) were dissolved in a mixed solvent of ethanol (50 mL) and water (10 mL), and the reaction solution was stirred at 80°C for 16 h. After monitoring the completion of the reaction, the reaction solution was concentrated, and the pH was adjusted to 5-6 with 2N dilute hydrochloric acid. The aqueous phase was extracted with ethyl acetate (30 mL x 3), and the combined organic phase was washed with saturated brine (20 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was directly used in the next step to obtain 4.3 g of the title compound as a yellow solid with a yield of 81%.

[0309] LCMS: m / z 286.00 [M+H] + .

[0310] Step 3: Preparation of 2-(N-isopropylacetylamino)thiophene-3-carboxylic acid (8c).

[0311] At room temperature, 2-(isopropylamino)thiophene-3-carboxylic acid (8b) (4.30 g, 0.0232 mol), triethylamine (7.03 g, 0.0696 mol) and DMAP (283 mg, 2.32 mmol) were dissolved in dichloromethane (50 mL), and acetic anhydride (4.73 g, 0.0464 mmol) was added dropwise. The reaction solution was stirred at room temperature for 16 h. After monitoring the reaction completion, water (40 mL) was added and the mixture was allowed to stand for stratification. The aqueous phase was extracted with dichloromethane (30 mL x 3), the combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain 3.50 g of the title compound as a black oil with a yield of 66%.

[0312] LCMS: m / z 228.00 [M+H] + .

[0313] Step 4: Preparation of N-(3-aminothiophen-2-yl)-N-isopropylacetamide (8d).

[0314] At room temperature, 2-(N-isopropylacetylamino)thiophene-3-carboxylic acid (8c) (3.30 g, 0.0145 mol) and triethylamine (4.40 g, 0.0436 mol) were dissolved in 1,4-dioxane (16 mL), and diphenylphosphoryl azide (5.99 g, 0.0218 mol) was added. The reaction solution was stirred at room temperature for 1 h, and then water (16 mL) was added and stirred at 100°C for 18 h. After monitoring the completion of the reaction, NaOH aqueous solution was added to adjust the pH to 8-9, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain 2.40 g of the title compound as a black oil with a yield of 83%.

[0315] LCMS: m / z 199.00 [M+H] + .

[0316] Step 5: Preparation of 3-isopropyl-2-methyl-3H-thieno[2,3-d]imidazole (8e).

[0317] At room temperature, N-(3-aminothiophen-2-yl)-N-isopropylacetamide (8d) (2.20 g, 11.1 mmol) was dissolved in toluene (22 mL), phosphorus oxychloride (1.86 g, 12.2 mmol) was added, and the reaction solution was stirred at 100°C for 16 h. After monitoring the completion of the reaction, the reaction solution was concentrated, 10% sodium carbonate aqueous solution (20 mL) was added, and extracted with ethyl acetate (20 mL x 3), the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (dichloromethane: methanol = 10: 1) to obtain 1.20 g of the title compound as a brown solid, with a yield of 60%.

[0318] LCMS: m / z 181.00 [M+H] + .

[0319] Step 6: Preparation of 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3H-thieno[2,3-d]imidazole (8f).

[0320] 3-Isopropyl-2-methyl-3H-thieno[2,3-d]imidazole (8e) (300 mg, 1.67 mmol) and 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxolane (776 mg, 4.17 mmol) were dissolved in tetrahydrofuran (15 mL) at -78°C, LDA (8.33 mL, 16.7 mmol) was added dropwise, and the reaction solution was stirred at -78°C for 3 h under nitrogen protection. The reaction solution was added with aqueous ammonium chloride solution and extracted with ethyl acetate (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain 400 mg of the title compound as a yellow oil with a yield of 78%.

[0321] LCMS: m / z 224.00 [M+H-82] + .

[0322] Step 7: Preparation of 5-(2-chloro-5-fluoropyrimidin-4-yl)-3-isopropyl-2-methyl-3H-thieno[2,3-d]imidazole (8 g).

[0323] 3-Isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3H-thieno[2,3-d]imidazole (8f) (400 mg, 1.78 mmol), potassium phosphate (757 mg, 3.57 mmol) and 2,4-dichloro-5-fluoropyrimidine (447 mg, 2.68 mmol) were dissolved in a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL) at room temperature, and Pd(dppf)Cl was added. 2 (130 mg, 0.178 mmol), the reaction solution was replaced with gas three times under nitrogen protection, and stirred at 100 ° C for 16 h. After monitoring the reaction completion, ethyl acetate (20 mL) was added to dilute the reaction solution, water (20 mL) was added, and the layers were separated by standing. The aqueous phase was extracted with ethyl acetate (20 mL x 3), and the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 1: 1) to obtain 110 mg of the title compound as a yellow oil, with a yield of 27%.

[0324] LCMS: m / z 311.00 [M+H] + .

[0325] Step 8: Preparation of (3S,4R)-4-((5-fluoro-4-(3-isopropyl-2-methyl-3H-thieno[2,3-d]imidazol-5-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (8).

[0326] 5-(2-Chloro-5-fluoropyrimidin-4-yl)-3-isopropyl-2-methyl-3H-thieno[2,3-d]imidazole (8 g) (110 mg, 0.355 mmol) was dissolved in N-methylpyrrolidone (2 mL) at room temperature, diisopropylethylamine (91.6 mg, 0.710 mmol) and (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (54.3 mg, 0.355 mmol) were added, and the reaction solution was stirred at 120 °C for 18 h. After monitoring the completion of the reaction, the reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 mL x 3), washed with saturated brine (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 25.0 mg of the title compound as a white solid, with a yield of 18%.

[0327] LC-MS: m / z 392.00 [M+H] + .

[0328] 1H NMR (400MHz, DMSO) δ8.32(d,J=3.6Hz,1H),7.75(d,J=1.4Hz,1H),7.02(d,J=7.6Hz,1H),4.91(d,J=4.5Hz,1H),4.69(dt,J=13.2,6.6Hz, 1H),3.82-3.72(m,4H),3.50(s,1H),3.37-3.31(m,2H),3.06(t,J=10.3Hz,1H),2.51(s,1H),1.99(d,J=10.9Hz,1H),1.49-1.44(m,7H).

[0329] Example 9: Preparation of (3S,4R)-4-((5-fluoro-4-(3-isopropyl-2,6-dimethyl-2H-thieno[3,2-c]pyrazol-5-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (9)

[0330]

[0331] Step 1: Preparation of 2,4-dimethylthiophene-3-amine (9a)

[0332] At room temperature, lithium aluminum hydride (17.6 g, 0.46 mol) and 1,4-dioxane (200 mL) were added to the reaction bottle, 3-amino-4-methylthiophene-2-carboxylic acid methyl ester (20.0 g, 0.11 mol) was added at 100°C in a nitrogen atmosphere, and the reaction was carried out at 100°C for 2 hours. After cooling to room temperature, methyl tert-butyl ether (100 mL), water (100 mL), and sodium hydroxide aqueous solution (15%, 100 mL) were added, and the mother liquor was collected by suction filtration and extracted with ethyl acetate (200 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 13.2 g of the title compound as a yellow oily liquid.

[0333] LCMS: m / z 128.1 [M+H] + .

[0334] Step 2: Preparation of 1-(6-methyl-1H-thieno[3,2-c]pyrazol-1-yl)ethan-1-one (9b)

[0335] At room temperature, 2,4-dimethylthiophene-3-amine (9a) (10.0 g, 78.74 mmol), potassium acetate (7.7 g, 78.57 mmol), and toluene (100 mL) were added to a reaction flask, and acetic anhydride (16.0 g, 156.86 mmol) and isoamyl nitrite (13.8 g, 117.95 mmol) were added dropwise at 80°C, and the temperature was slowly raised to 95°C and stirred for 2 hours. After the reaction was completed, the pH was adjusted to 8 with saturated sodium bicarbonate, and ethyl acetate (200 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 6.5 g of the title compound as a brown oily liquid with a yield of 46.1%.

[0336] LCMS: m / z 181.0 [M+H] + .

[0337] Step 3: Preparation of 6-methyl-1H-thieno[3,2-c]pyrazole (9c)

[0338] At room temperature, 1-(6-methyl-1H-thieno[3,2-c]pyrazol-1-yl)ethane-1-one (9b) (5.5 g, 30.55 mmol), concentrated hydrochloric acid (25 mL, 0.30 mol), ethanol (20 mL) and water (20 mL) were added to a reaction flask and stirred at 60°C for 2 hours. After the reaction, the pH was adjusted to 8 with saturated sodium bicarbonate, and ethyl acetate (100 mL) and water (100 mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 3.8 g of the title compound as a dark brown solid.

[0339] LCMS: m / z 139.0 [M+H] + .

[0340] Step 4: Preparation of 3-iodo-6-methyl-1H-thieno[3,2-c]pyrazole (9d)

[0341] At room temperature, 6-methyl-1H-thieno[3,2-c]pyrazole (9c) (3.8 g, 27.54 mmol), potassium carbonate (8.3 g, 60.14 mmol), iodine (7.6 g, 29.94 mmol) and methanol (40 mL) were added to a reaction flask and stirred at room temperature overnight. After the reaction was completed, ethyl acetate (100 mL) and saturated sodium thiosulfate (100 mL) were added for extraction, the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 5.3 g of the title compound as a brown solid, with a yield of 72.9%.

[0342] LCMS: m / z 264.90 [M+H]+ .

[0343] Step 5: Preparation of 3-iodo-2,6-dimethyl-2H-thieno[3,2-c]pyrazole (9e)

[0344] At room temperature, 3-iodo-6-methyl-1H-thieno[3,2-c]pyrazole (9d) (5.0 g, 18.94 mmol), trimethyloxonium tetrafluoroborate (2.9 g, 19.59 mmol), sodium bicarbonate (3.1 g, 37.50 mmol) and dichloromethane (50 mL) were added to a reaction flask and stirred at room temperature for 2 hours. After the reaction was completed, ethyl acetate (50 mL) and water (50 mL) were added for extraction. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 5.0 g of the title compound as a brown-yellow solid with a yield of 95.2%.

[0345] LCMS: m / z 278.80 [M+H] + .

[0346] Step 6: Preparation of 2,6-dimethyl-3-(prop-1-en-2-yl)-2H-thieno[3,2-c]pyrazole (9f)

[0347] At room temperature, 3-iodo-2,6-dimethyl-2H-thieno[3,2-c]pyrazole (9e) (5.0 g, 17.99 mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (4.5 g, 26.78 mmol), Pd(dppf)Cl 2 (1.3g, 1.77mmol), potassium phosphate (11.4g, 53.77mmol), 1,4-dioxane (100mL) and water (10mL) were added to the reaction bottle, and the reaction was stirred at 100°C in a nitrogen atmosphere overnight. After the reaction was completed, it was cooled to room temperature, and ethyl acetate (200mL) and water (100mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 6.5g of the dark brown liquid title compound with a yield of 94.2%.

[0348] LCMS: m / z 193.0 [M+H] + .

[0349] Step 7: Preparation of 3-isopropyl-2,6-dimethyl-2H-thieno[3,2-c]pyrazole (9 g)

[0350] At room temperature, 2,6-dimethyl-3-(prop-1-en-2-yl)-2H-thieno[3,2-c]pyrazole (9f) (6.5 g, 33.85 mmol), palladium on carbon (650 mg), anhydrous methanol (60 mL) and acetic acid (10 mL) were added to a reaction flask, and the mixture was stirred at room temperature under hydrogen atmosphere overnight. After the reaction was completed, the mother liquor was collected by suction filtration and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 5.0 g of the title compound as a brown liquid with a yield of 76.2%.

[0351] LCMS: m / z 195.0 [M+H] + .

[0352] Step 8: Preparation of 3-isopropyl-2,6-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-thieno[3,2-c]pyrazole (9h)

[0353] At room temperature, 3-isopropyl-2,6-dimethyl-2H-thieno[3,2-c]pyrazole (9g) (1.0g, 5.15mmol), 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborane (2.4g, 12.88mmol) and anhydrous tetrahydrofuran (10mL) were added to the reaction bottle, and lithium diisopropylamide (2N, 12.8mL, 25.77mmol) was added dropwise at -78°C under nitrogen atmosphere. The reaction was stirred at this temperature for 4 hours. After slowly returning to room temperature, ethyl acetate (50mL) and saturated ammonium chloride (50mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 300.0mg of the light yellow liquid title compound, with a yield of 18.7%.

[0354] LCMS: m / z 321.1 [M+H] + .

[0355] Step 9: Preparation of 5-(2-chloro-5-fluoropyrimidin-4-yl)-3-isopropyl-2,6-dimethyl-2H-thieno[3,2-c]pyrazole (9i)

[0356] At room temperature, 3-isopropyl-2,6-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-thieno[3,2-c]pyrazole (9h) (500.0 mg, 1.56 mmol), 2,4-dichloro-5-fluoropyrimidine (310.4 mg, 1.87 mmol), Pd(dppf)Cl 2(136.7 mg, 0.18 mmol), potassium phosphate (661.4 mg, 3.12 mmol), 1,4-dioxane (5 mL) and water (1 mL) were added to the reaction bottle and microwaved at 100 ° C for 1 hour under nitrogen atmosphere. After the reaction was completed, it was cooled to room temperature and extracted with ethyl acetate (50 mL) and water (50 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 4:1) to obtain 350 mg of the light yellow liquid title compound with a yield of 69.1%.

[0357] LCMS: m / z 325.0 [M+H] + .

[0358] Step 10: Preparation of (3S,4R)-4-((5-fluoro-4-(3-isopropyl-2,6-dimethyl-2H-thieno[3,2-c]pyrazol-5-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (9)

[0359] At room temperature, 5-(2-chloro-5-fluoropyrimidin-4-yl)-3-isopropyl-2,6-dimethyl-2H-thieno[3,2-c]pyrazole (9i) (100.0 mg, 0.30 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (56.8 mg, 0.37 mmol), diisopropylethylamine (79.6 mg, 0.61 mmol) and N-methylpyrrolidone (1 mL) were added to a reaction bottle and reacted at 130°C for 3 hours under microwave. After cooling to room temperature, ethyl acetate (20 mL) and water (5 mL) were added. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 24.0 mg of the title compound as a light yellow solid, with a yield of 19.2%.

[0360] LCMS: m / z 406.1 [M+H] + .

[0361] 1H NMR (400MHz, DMSO) δ8.43(d,J=3.0Hz,1H),7.20(d,J=7.8Hz,1H),4.90(d,J=5.3Hz,1H),3.99(s,3H),3.84-3.73(m,3H),3.53-3.47(m,1H),3. 38-3.33(m,1H),3.30-3.27(m,1H),3.07-3.01(m,1H),2.41(d,J=2.9Hz ,3H),1.97(d,J=11.7Hz,1H),1.51-1.42(m,1H),1.31(d,J=6.8Hz,6H).

[0362] Example 10: Preparation of 2-(2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)-5-methylpyrimidin-4-yl)-7-isopropyl-5-methylthieno[3,2-c]pyridin-4(5H)-one (10)

[0363]

[0364] Step 1: Preparation of 1-(5-bromothiophen-2-yl)-2-methylpropan-1-one (10a)

[0365] At room temperature, 2-bromothiophene (10.0 g, 6.17 mmol), aluminum chloride (9.0 g, 6.79 mmol), and dichloromethane (100 mL) were added to the reaction flask, and isobutyryl chloride (7.20 g, 6.79 mmol) was added dropwise at 0°C, and the temperature was slowly raised to room temperature and stirred overnight. After the reaction was completed, dichloromethane (100 mL) and water (100 mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 10:1) to obtain 13.1 g of the title compound as a light yellow oily liquid with a yield of 90.9%.

[0366] LCMS: m / z 234.90 [M+H] + .

[0367] Step 2: Preparation of ethyl 3-(5-bromothiophen-2-yl)-4-methylpent-2-enoate (10b)

[0368] At room temperature, triethyl phosphonoacetate (9.6 g, 43.20 mmol) and tetrahydrofuran (50 mL) were added to the reaction bottle, sodium hydride (1.7 g, 43.20 mmol) was added in batches at 0°C, the temperature was slowly raised to room temperature and stirred for 1 hour, 1-(5-bromothiophen-2-yl)-2-methylpropan-1-one (10a) (5.0 g, 21.55 mmol) was added, and the reaction was stirred at room temperature overnight under nitrogen atmosphere. After the reaction was completed, water (100 mL) was added to quench the reaction, and ethyl acetate (100 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 5.8 g of the title compound as a yellow-brown oily liquid, with a yield of 89.2%.

[0369] LCMS: m / z 304.90 [M+H] + .

[0370] Step 3: Preparation of 3-(5-bromothiophen-2-yl)-4-methylpent-2-enoic acid (10c)

[0371] At room temperature, ethyl 3-(5-bromothiophen-2-yl)-4-methylpent-2-enoate (10b) (5.8 g, 19.2 mmol), sodium hydroxide (3.0 g, 76.8 mmol), methanol (60 mL), and water (15 mL) were added to a reaction flask and stirred at 65°C for 1 hour. After the reaction, a large amount of methanol was concentrated, and the residue was adjusted to pH ≤ 3 with dilute hydrochloric acid (2N, 60 mL). Ethyl acetate (100 mL) and water (50 mL) were added for extraction, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 4.2 g of the title compound as an off-white solid.

[0372] LCMS: m / z 274.90 [M+H] + .

[0373] Step 4: Preparation of (2-bromo-7-isopropylthieno[3,2-c]pyridin-4(5H)-one (10d)

[0374] At room temperature, 3-(5-bromothiophen-2-yl)-4-methylpent-2-enoic acid (10c) (2.3 g, 8.39 mmol), triethylamine (2.5 g, 25.1 mmol), diphenylphosphoryl azide (3.4 g, 12.59 mmol), and diphenyl ether (20 mL) were added to a reaction flask, and the mixture was stirred in a microwave at 180°C for 30 minutes under a nitrogen atmosphere. After cooling to room temperature, the mixture was concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 10:1) to obtain 2.1 g of the title compound as a dark brown oily liquid with a yield of 46.7%.

[0375] LCMS: m / z 273.90 [M+H] + .

[0376] Step 5: Preparation of 2-bromo-7-isopropyl-5-methylthieno[3,2-c]pyridin-4(5H)-one (10e)

[0377] At room temperature, (2-bromo-7-isopropylthieno[3,2-c]pyridin-4(5H)-one (10d) (2.0 g, 7.38 mmol), iodomethane (2.0 g, 14.76 mmol), cesium carbonate (4.8 g, 14.76 mmol), and N,N-dimethylformamide (20 mL) were added to a reaction flask and stirred at room temperature overnight. After the reaction, ethyl acetate (50 mL) and water (50 mL) were added for extraction. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:2) to obtain 1.4 g of the title compound as a dark brown waxy solid in a yield of 66.7%.

[0378] LCMS: m / z 287.90 [M+H] + .

[0379] Step 6: Preparation of (7-isopropyl-5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridin-2-yl)boronic acid (10f)

[0380] At room temperature, 2-bromo-7-isopropyl-5-methylthieno[3,2-c]pyridin-4(5H)-one (10e) (1.4 g, 4.87 mmol), bis(boronic acid pinacol) (1.8 g, 7.28 mmol), Pd(dppf)Cl 2 (358.4 mg, 0.49 mmol), potassium acetate (1.4 g, 14.7 mmol), and 1,4-dioxane (10 mL) were added to the reaction flask and stirred at 100 ° C in a nitrogen atmosphere overnight. After cooling to room temperature, ethyl acetate (50 mL) and water (50 mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 1.3 g of the title compound as a brown-black solid.

[0381] LCMS: m / z 252.0 [M+H] + .

[0382] Step 7: Preparation of 2-(2-chloro-5-methylpyrimidin-4-yl)-7-isopropyl-5-methylthieno[3,2-c]pyridin-4(5H)-one (10 g)

[0383] At room temperature, (7-isopropyl-5-methyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridin-2-yl)boronic acid (10f) (1.3 g, 5.18 mmol), 2,4-dichloro-5-methylpyrimidine (1.25 g, 7.76 mmol), Pd(dppf)Cl 2 (372.8 mg, 0.51 mmol), potassium phosphate (3.3 g, 15.53 mmol), 1,4-dioxane (10 mL), and water (1 mL) were added to the reaction flask and stirred at 100 ° C in a nitrogen atmosphere overnight. After cooling to room temperature, ethyl acetate (50 mL) and water (50 mL) were added for extraction. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: dichloromethane: methanol = 5:1) to obtain 720 mg of the title compound as a black solid with a yield of 42.3%.

[0384] LCMS: m / z 334.0 [M+H] + .

[0385] Step 8: Preparation of 2-(2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)-5-methylpyrimidin-4-yl)-7-isopropyl-5-methylthieno[3,2-c]pyridin-4(5H)-one (10)

[0386] At room temperature, 2-(2-chloro-5-methylpyrimidin-4-yl)-7-isopropyl-5-methylthieno[3,2-c]pyridin-4(5H)-one (10 g) (100 mg, 0.30 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (50.8 mg, 0.30 mmol), diisopropylethylamine (77.4 mg, 0.60 mmol), N-methylpyrrolidone (NMP) (1 mL) were added to the reaction bottle, and the reaction was stirred at 120°C overnight. After cooling to room temperature, ethyl acetate (20 mL) and water (5 mL) were added. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 27.0 mg of the light yellow solid title compound, yield 21.6%

[0387] LCMS: m / z 415.1 [M+H] + .

[0388] 1H NMR (400MHz, DMSO) δ8.23(s,1H),7.95(s,1H),7.50(s,1H),6.94(d,J=7.5Hz,1H),4.92(d,J=5.3Hz,1H),3.87-3.75(m,3H),3.56-3.49(m ,4H),3.38-3.34(m,1H),3.12-3.03(m,1H),2.95-2.88(m,1H),2.39(s,3H),2.09-1.95(m,1H),1.51-1.42(m,1H),1.32(d,J=6.9Hz,6H).

[0389] Example 11: Preparation of (3S,4R)-4-((5-fluoro-4-(3-isopropyl-2-methyl-2H-thieno[3,2-c]pyrazol-5-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (11)

[0390]

[0391] Step 1: Preparation of 2-methylthiophene-3-amine (11a)

[0392] At room temperature, lithium aluminum hydride (9.6 g, 0.25 mol) and 1,4-dioxane (100 mL) were added to the reaction bottle, 3-aminothiophene-2-carboxylic acid methyl ester (10.0 g, 0.63 mol) was added at 100°C in a nitrogen atmosphere, and the reaction was carried out at 100°C for 2 hours. After cooling to room temperature, methyl tert-butyl ether (100 mL), water (100 mL), and sodium hydroxide aqueous solution (15%, 100 mL) were added, and the mother liquor was collected by suction filtration and extracted with ethyl acetate (200 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 5.0 g of the title compound as a yellow oily liquid.

[0393] LCMS: m / z 114.0 [M+H] + .

[0394] Step 2: Preparation of 1-(1H-thieno[3,2-c]pyrazol-1-yl)ethan-1-one (11b)

[0395] At room temperature, 2-methylthiophene-3-amine (11a) (5.0 g, 44.24 mmol), potassium acetate (9.0 g, 88.23 mmol), and toluene (50 mL) were added to a reaction flask, and acetic anhydride (4.3 g, 43.87 mmol) and isoamyl nitrite (7.7 g, 65.81 mmol) were added dropwise at 80 °C, and the temperature was slowly raised to 95 °C and stirred for 1 hour. After the reaction was completed, saturated sodium bicarbonate was used to adjust the pH to 8, and ethyl acetate (200 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 4.2 g of the title compound as a brown oily liquid with a yield of 76.6%.

[0396] LCMS: m / z 167.0 [M+H] + .

[0397] Step 3: Preparation of 1H-thieno[3,2-c]pyrazole (11c)

[0398] At room temperature, 1-(1H-thieno[3,2-c]pyrazol-1-yl)ethane-1-one (11b) (4.2 g, 25.30 mmol), concentrated hydrochloric acid (21 mL, 0.25 mol), ethanol (20 mL) and water (20 mL) were added to a reaction flask and stirred at 60°C for 2 hours. After the reaction, saturated sodium bicarbonate was used to adjust the pH to 8, and ethyl acetate (100 mL) and water (100 mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 2.0 g of the title compound as a dark brown solid.

[0399] LCMS: m / z 125.0 [M+H] + .

[0400] Step 4: Preparation of 3-iodo-1H-thieno[3,2-c]pyrazole (11d)

[0401] At room temperature, 1H-thieno[3,2-c]pyrazole (11c) (2.0 g, 16.1 mmol), potassium carbonate (4.9 g, 35.50 mmol), iodine (4.5 g, 17.73 mmol) and methanol (20 mL) were added to a reaction flask and stirred at room temperature overnight. After the reaction was completed, ethyl acetate (100 mL) and saturated sodium thiosulfate (100 mL) were added for extraction, the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 4.0 g of the title compound as a brown solid, with a yield of 99.2%.

[0402] LCMS: m / z 250.8 [M+H] + .

[0403] Step 5: Preparation of 3-iodo-2-methyl-2H-thieno[3,2-c]pyrazole (11e)

[0404] At room temperature, 3-iodo-1H-thieno[3,2-c]pyrazole (11d) (4.0 g, 16.00 mmol), trimethyloxonium tetrafluoroborate (2.4 g, 16.21 mmol), sodium bicarbonate (2.6 g, 30.95 mmol) and dichloromethane (40 mL) were added to a reaction flask and stirred at room temperature overnight. After the reaction was completed, ethyl acetate (100 mL) and water (100 mL) were added for extraction. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 3.2 g of the title compound as a brown-yellow solid with a yield of 76.2%.

[0405] LCMS: m / z 264.90 [M+H] + .

[0406] Step 6: Preparation of 2-methyl-3-(propyl-1-en-2-yl)-2H-thieno[3,2-c]pyrazole (11f)

[0407] At room temperature, 3-iodo-2-methyl-2H-thieno[3,2-c]pyrazole (13e) (3.2 g, 12.12 mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (3.0 g, 18.18 mmol), Pd(dppf)Cl 2 (884.5mg, 1.21mmol), potassium phosphate (7.7g, 36.36mmol), 1,4-dioxane (30mL) and water (5mL) were added to the reaction bottle, and the reaction was stirred at 100°C in a nitrogen atmosphere overnight. After the reaction was completed, it was cooled to room temperature, and ethyl acetate (100mL) and water (100mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 1.8g of the light yellow liquid title compound, with a yield of 66.6%.

[0408] LCMS: m / z 179.0 [M+H] + .

[0409] Step 7: Preparation of 3-isopropyl-2-methyl-2H-thieno[3,2-c]pyrazole (11 g)

[0410] At room temperature, 2-methyl-3-(propyl-1-en-2-yl)-2H-thieno[3,2-c]pyrazole (11f) (1.7 g, 9.55 mmol), platinum dioxide (100.0 mg), and anhydrous methanol (20 mL) were added to a reaction flask, and the mixture was stirred at room temperature under a hydrogen atmosphere overnight. After the reaction was completed, the mother liquor was collected by suction filtration and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 1.5 g of the light yellow liquid title compound, with a yield of 88.2%.

[0411] LCMS: m / z 181.0 [M+H] + .

[0412] Step 8: Preparation of (3-isopropyl-2-methyl-2H-thieno[3,2-c]pyrazol-5-yl)boronic acid (11h)

[0413] At room temperature, 3-isopropyl-2-methyl-2H-thieno[3,2-c]pyrazole (11 g) (600.0 mg, 3.33 mmol), 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborane (1.5 g, 8.33 mmol) and anhydrous tetrahydrofuran (6 mL) were added to a reaction flask, and lithium diisopropylamide (2N, 8.3 mL, 16.66 mmol) was added dropwise at -78 °C under a nitrogen atmosphere. The reaction was stirred at this temperature for 4 hours. After slowly returning to room temperature, ethyl acetate (50 mL) and saturated ammonium chloride (50 mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 700.0 mg of the light yellow liquid title compound with a yield of 93.8%.

[0414] LCMS: m / z 225.1 [M+H] + .

[0415] Step 9: Preparation of 5-(2-chloro-5-fluoropyrimidin-4-yl)-3-isopropyl-2-methyl-2H-thieno[3,2-c]pyrazole (11i)

[0416] At room temperature, (3-isopropyl-2-methyl-2H-thieno[3,2-c]pyrazol-5-yl)boronic acid (11h) (500.0 mg, 2.23 mmol), 2,4-dichloro-5-fluoropyrimidine (443.2 mg, 2.67 mmol), Pd(dppf)Cl 2(160.8mg, 0.22mmol), potassium phosphate (945.5mg, 4.46mmol), 1,4-dioxane (5mL) and water (1mL) were added to the reaction bottle and reacted overnight at 100°C in a nitrogen atmosphere. After the reaction was completed, it was cooled to room temperature and extracted with ethyl acetate (50mL) and water (50mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 4:1) to obtain 360mg of the title compound as a dark brown solid with a yield of 52.0%.

[0417] LCMS: m / z 311.0 [M+H] + .

[0418] Step 10: Preparation of (3S,4R)-4-((5-fluoro-4-(3-isopropyl-2-methyl-2H-thieno[3,2-c]pyrazol-5-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (13)

[0419] At room temperature, 5-(2-chloro-5-fluoropyrimidin-4-yl)-3-isopropyl-2-methyl-2H-thieno[3,2-c]pyrazole (11i) (100.0 mg, 0.32 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (59.2 mg, 0.38 mmol), diisopropylethylamine (83.2 mg, 0.64 mmol) and N-methylpyrrolidone (1 mL) were added to a reaction bottle and reacted at 130°C for 3 hours under microwave. After cooling to room temperature, ethyl acetate (20 mL) and water (20 mL) were added. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 58.0 mg of the title compound as a light yellow solid, with a yield of 46.0%.

[0420] LCMS: m / z 392.1 [M+H] + .

[0421] 1H NMR (400MHz, DMSO) δ8.43(d,J=3.3Hz,1H),7.78(s,1H),7.17(d,J=7.7Hz,1H),4.91(d,J=5.3Hz,1H),4.00(s,3H),3.89-3.67(m ,3H),3.59-3.41(m,1H),3.38-3.33(m,2H),3.06(t,J=10.4Hz,1H),2.04-1.94(m,1H),1.54-1.40(m,1H),1.34(d,J=6.8Hz,6H).

[0422] Example 12: Preparation of (3S,4R)-4-((4-(2,6-dimethyl-3-(1,1,1-trifluoro-2-hydroxypropan-2-yl)-2H-thieno[3,2-c]pyrazol-5-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (12)

[0423]

[0424] Step 1: Preparation of 2,4-dimethylthiophene-3-amine (12a)

[0425] At room temperature, lithium aluminum tetrahydride (4.44 g, 116.96 mmol) and 1,4-dioxane (100 mL) were placed in a reaction bottle, nitrogen was replaced and the temperature was raised to 80°C, and a mixture of 3-amino-4-methylthiophene-2-carboxylic acid methyl ester (5.0 g, 29.24 mmol) and 1,4-dioxane (40 mL) was slowly added dropwise, and the temperature was raised to 100°C, and the reaction was stirred for 2 hours. After the reaction was completed, water was added to quench the reaction, filtered with diatomaceous earth, the filtrate was collected, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), dried with anhydrous sodium sulfate, and condensed under reduced pressure, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 2.86 g of the title product as a yellow oily liquid, with a yield of 77.0%.

[0426] LCMS: m / z 128.0 [M+H] + .

[0427] Step 2: Preparation of 1-(6-methyl-1H-thieno[3,2-c]pyrazol-1-yl)ethan-1-one (12b)

[0428] At room temperature, 2,4-dimethylthiophene-3-amine (2.86 g, 22.44 mmol), potassium acetate (2.20 g, 22.44 mmol), acetic anhydride (4.58 g, 44.88 mmol) and toluene (30 ml) were added to the reaction bottle, the temperature was raised to 80 ° C, and the reaction was stirred for 10 min. Isoamyl nitrite (3.94 g, 33.66 mmol) was added and the temperature was raised to 95 ° C, and the reaction was stirred for 2 h. After the reaction was completed, the reaction was quenched with a saturated sodium bicarbonate aqueous solution, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was dried and concentrated, dried over anhydrous sodium sulfate, and condensed under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 2.53 g of the title product as a yellow oily liquid with a yield of 62.6%.

[0429] LCMS: m / z 181.0 [M+H] + .

[0430] Step 3: Preparation of 6-methyl-1H-thieno[3,2-c]pyrazole (12c)

[0431] At room temperature, 1-(6-methyl-1H-thieno[3,2-c]pyrazol-1-yl)ethane-1-one (2.53 g, 14.0 mmol), concentrated hydrochloric acid (11.68 mL, 140.0 mmol), anhydrous ethanol (10 mL) and water (10 mL) were placed in a reaction bottle, heated to 60°C, and stirred for 2 h. After the reaction, saturated sodium bicarbonate aqueous solution was added to adjust the pH to 7-8, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 1.07 g of the title product as a yellow solid with a yield of 55.4%.

[0432] LCMS: m / z 139.0 [M+H] + .

[0433] Step 4: Preparation of 3-iodo-6-methyl-1H-thieno[3,2-c]pyrazole (12d)

[0434] At room temperature, 6-methyl-1H-thieno[3,2-c]pyrazole (1.07 g, 7.78 mmol), potassium carbonate (2.15 g, 15.57 mmol), iodine (2.17 g, 8.56 mmol) and anhydrous methanol (10 mL) were placed in a reaction flask and stirred at room temperature overnight. After the reaction was completed, a saturated sodium thiosulfate aqueous solution was added to quench the reaction, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 1.98 g of the title product as a white solid with a yield of 96.4%.

[0435] LCMS: m / z 264.9 [M+H] + .

[0436] Step 5: Preparation of 3-iodo-2,6-dimethyl-2H-thieno[3,2-c]pyrazole (12e).

[0437] At room temperature, 3-iodo-6-methyl-1H-thieno[3,2-c]pyrazole (1.98 g, 7.50 mmol) and anhydrous tetrahydrofuran (30 mL) were placed in a reaction bottle, nitrogen was replaced, the temperature was lowered to 0°C, potassium tert-butoxide (9.0 mL, 9.0 mmol) was added, the reaction was stirred at room temperature for 10 min, iodomethane (1.28 g, 9.0 mmol) was added, and the reaction was stirred for 2 h. After the reaction was completed, saturated aqueous ammonium chloride solution was added to quench the reaction, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 1.60 g of the title product as a white solid with a yield of 76.8%.

[0438] LCMS: m / z 278.9 [M+H] + .

[0439] Step 6: Preparation of 1-(2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)ethan-1-one (12f).

[0440] At room temperature, 3-iodo-2,6-dimethyl-2H-thieno[3,2-c]pyrazole (3.41 g, 12.27 mmol), tributyl(1-ethoxyethylene)tin (6.65 g, 18.41 mmol), tetrakis(triphenylphosphine)palladium and toluene (30 mL) were placed in a reaction bottle, nitrogen was replaced, the temperature was raised to 100°C, and the reaction was stirred overnight. 10% potassium fluoride solution was added to quench the reaction, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure, anhydrous tetrahydrofuran (30 mL) and 1N hydrochloric acid (30 mL) were added, and the reaction was stirred at room temperature overnight. After the reaction, saturated sodium bicarbonate aqueous solution was added to adjust the pH to 7-8, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 1.35 g of the title product as a brown solid with a yield of 56.7%.

[0441] LCMS: m / z 195.1 [M+H] + .

[0442] Step 7: Preparation of 2-(2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)-1,1,1-trifluoropropan-2-ol (12 g).

[0443] At room temperature, 1-(2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)ethane-1-one (378 mg, 1.95 mmol) and anhydrous tetrahydrofuran (10 mL) were placed in a reaction bottle, nitrogen was replaced, the temperature was lowered to 0°C, (trifluoromethyl)trimethylsilane (1.94 g, 13.65 mmol) and tetrabutylammonium fluoride (2.93 mL, 2.93 mmol) were added, and the reaction was stirred for 1 hour. After the reaction was completed, saturated aqueous ammonium chloride solution was added to quench the reaction, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 568 mg of the title product as a yellow solid with a yield of 90.8%.

[0444] LCMS: m / z 265.1 [M+H] + .

[0445] Step 8: Preparation of (2,6-dimethyl-3-(1,1,1-trifluoro-2-hydroxypropyl-2-yl)-2H-thieno[3,2-c]pyrazol-5-yl)boronic acid (12h).

[0446] At room temperature, 2-(2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)-1,1,1-trifluoropropan-2-ol (388 mg, 1.47 mmol), 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborane (547.1 mg, 2.94 mmol) and anhydrous tetrahydrofuran (10 mL) were placed in a reaction bottle, nitrogen was replaced, the temperature was lowered to -78°C, LDA (2.94 mL, 7.35 mmol) was added, and the reaction was stirred overnight. After the reaction was completed, saturated aqueous ammonium chloride solution was added to quench the reaction, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 452 mg of the title product (crude product) as an orange solid.

[0447] LCMS: m / z 309.1[M+H] + .

[0448] Step 9: Preparation of 2-(5-(2-chloro-5-fluoropyrimidin-4-yl)-2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)-1,1,1-trifluoropropan-2-ol (12i).

[0449] At room temperature, (2,6-dimethyl-3-(1,1,1-trifluoro-2-hydroxypropyl-2-yl)-2H-thieno[3,2-c]pyrazol-5-yl)boronic acid (452 ​​mg, 1.47 mmol), 2,4-dichloro-5-fluoropyrimidine (366 mg, 2.21 mmol), Pd(dppf)Cl 2 (107mg, 0.147mmol), potassium carbonate (608mg, 4.41mmol), 1,4-dioxane (10mL) and water (3mL) were placed in a reaction flask, nitrogen was replaced, the temperature was raised to 100°C, and the reaction was stirred overnight. After the reaction was completed, the filtrate was filtered with diatomaceous earth, ethyl acetate (50mL) and water (20mL) were added to take the organic phase, the organic phase was washed with water (30mL*3), dried with anhydrous sodium sulfate, and condensed under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 120mg of the title product as a yellow liquid with a yield of 30.5%.

[0450] LCMS: m / z 395.0 [M+H] + .

[0451] Step 10: Preparation of (3S,4R)-4-((4-(2,6-dimethyl-3-(1,1,1-trifluoro-2-hydroxypropan-2-yl)-2H-thieno[3,2-c]pyrazol-5-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (12).

[0452] At room temperature, 2-(5-(2-chloro-5-fluoropyrimidin-4-yl)-2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)-1,1,1-trifluoropropan-2-ol (100 mg, 0.25 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (39.1 mg, 0.25 mmol), DIEA (69 mg, 0.50 mmol) and NMP (2 mL) were added to a reaction bottle, the temperature was raised to 120°C, and the reaction was stirred overnight. After the reaction, the filtrate was collected by diatomaceous earth filtration, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was dried and concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 36 mg of the light yellow solid title compound, with a yield of 30.3%.

[0453] LCMS: m / z 476.1[M+H] + .

[0454] 1H NMR (400MHz, CDCl3) δ8.24(d,J=2.1Hz,1H),5.33(d,J=5.8Hz,1H),4.16(s,3H),4.05(dd ,J=11.4,4.8Hz,1H),3.98(dd,J=11.8,3.4Hz,2H),3.85(ddd,J=15.5,15.0,9.6Hz,1H),3 .61(td,J=9.4,4.9Hz,1H),3.47(td,J=11.8,2.1Hz,1H),3.20(dd,J=11.3,9.9Hz,1H),2. 54(d,J=3.3Hz,3H),2.12–2.02(m,1H),1.80(s,3H),1.68(ddd,J=24.6,11.8,4.7Hz,2H).

[0455] Example 13: Preparation of (3S,4R)-4-((5-fluoro-4-(3-(2-hydroxypropan-2-yl)-2,6-dimethyl-2H-thieno[3,2-c]pyrazol-5-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (13)

[0456]

[0457] Step 1: Preparation of 2-(2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)propan-2-ol (13a)

[0458] At room temperature, 1-(2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)ethane-1-one (909 mg, 4.68 mmol) and anhydrous tetrahydrofuran (15 mL) were placed in a reaction bottle, nitrogen was replaced and the temperature was lowered to 0°C, methylmagnesium bromide (14 mL, 14.0 mmol) was slowly added dropwise, and the reaction was stirred for 16 hours. After the reaction was completed, saturated aqueous ammonium chloride solution was added dropwise to quench the reaction, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), dried over anhydrous sodium sulfate, and condensed under reduced pressure, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 560 mg of the title product as a yellow solid, with a yield of 57.0%.

[0459] LCMS: m / z 211.1 [M+H] + .

[0460] Step 2: Preparation of 2-(2,6-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-thieno[3,2-c]pyrazol-3-yl)propan-2-ol (13b)

[0461] At room temperature, 2-(2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)propan-2-ol (560 mg, 2.66 mmol), 2-(tert-butoxy)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (989.5 mg, 5.32 mmol) and anhydrous tetrahydrofuran (20 mL) were added to the reaction bottle, nitrogen was replaced and the temperature was lowered to -78°C, the reaction was stirred for 10 min, LDA (6.65 mL, 13.30 mmol) was added and the reaction was stirred for 16 h. After the reaction was completed, the reaction was quenched with saturated aqueous ammonium chloride solution, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was dried and concentrated, dried over anhydrous sodium sulfate, and condensed under reduced pressure, and the crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 760 mg of the title product as a light yellow solid, with a yield of 84.7%.

[0462] LCMS: m / z 337.2 [M+H] + .

[0463] Step 3: Preparation of 2-(5-(2-chloro-5-fluoropyrimidin-4-yl)-2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)propan-2-ol (13c)

[0464] At room temperature, 2-(2,6-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-thieno[3,2-c]pyrazol-3-yl)propan-2-ol (1.40 g, 4.17 mmol), 2,4-dichloro-5-fluoropyrimidine (1.04 g, 6.25 mmol), Pd(dppf)Cl 2 (305mg, 0.417mmol), potassium carbonate (1.73g, 12.5mmol), 1,4-dioxane (20mL) and water (6mL) were placed in a reaction flask, nitrogen was replaced, the temperature was raised to 100°C, and the reaction was stirred overnight. After the reaction was completed, the filtrate was filtered with diatomaceous earth, ethyl acetate (50mL) and water (20mL) were added to take the organic phase, the organic phase was washed with water (30mL*3), dried with anhydrous sodium sulfate, and condensed under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 1.12g of the title product as a yellow liquid with a yield of 78.8%.

[0465] LCMS: m / z 341.0 [M+H] + .

[0466] Step 4: Preparation of (3S,4R)-4-((5-fluoro-4-(3-(2-hydroxypropan-2-yl)-2,6-dimethyl-2H-thieno[3,2-c]pyrazol-5-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (13)

[0467] At room temperature, 2-(5-(2-chloro-5-fluoropyrimidin-4-yl)-2,6-dimethyl-2H-thieno[3,2-c]pyrazol-3-yl)propan-2-ol (120 mg, 0.36 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (55.1 mg, 0.36 mmol), DIEA (93.1 mg, 0.72 mmol) and NMP (2 mL) were added to a reaction bottle, the temperature was raised to 120°C, and the reaction was stirred overnight. After the reaction, the filtrate was collected by diatomaceous earth filtration, ethyl acetate (50 mL) and water (20 mL) were added to take the organic phase, the organic phase was washed with water (30 mL*3), the organic phase was dried and concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 45 mg of the light yellow solid title compound, with a yield of 29.7%.

[0468] LCMS: m / z 422.2 [M+H] + .

[0469] 1H NMR (400MHz, DMSO) δ8.41(t,J=14.9Hz,1H),7.21(d,J=7.8Hz,1H),5.29(d,J=19.1Hz,1H),4.91(d,J=5.3Hz,1H),4.06(s,3H),4.00–3.92(m,1H), 3.87–3.69(m,3H),3.50(ddd,J=14.4,9.6,5.0Hz,1H),3.11–2.97(m,1H) ,2.54(d,J=2.7Hz,3H),1.95(t,J=14.1Hz,1H),1.49(s,6H),1.46(s,1H).

[0470] Example 14: Preparation of (3S,4R)-4-((5-fluoro-4-(7-isopropyl-3-(trifluoromethyl)thieno[3,2-b]pyridin-2-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (14)

[0471]

[0472] Step 1: Preparation of 7-(prop-1-en-2-yl)thieno[3,2-b]pyridine (14a)

[0473] At room temperature, 7-chlorothieno[3,2-b]pyridine (6.00 g, 35.5 mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (9.00 g, 53.2 mmol), Pd(dppf)Cl 2 (2.60g, 3.50mmol), potassium phosphate (22.5g, 0.106mol), 1,4-dioxane (60mL), and water (18mL) were added to the reaction flask, and the reaction was stirred at 100°C in a nitrogen atmosphere overnight. After cooling to room temperature, ethyl acetate (300mL) and water (300mL) were added for extraction. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 5.80g of the title compound as a light yellow oily liquid with a yield of 93.5%.

[0474] LCMS: m / z 176.0 [M+H] + .

[0475] Step 2: Preparation of 7-isopropylthieno[3,2-b]pyridine (14b)

[0476] At room temperature, 7-(prop-1-en-2-yl)thieno[3,2-b]pyridine (5.80 g, 33.1 mmol) (14a), 5% palladium on carbon (580 mg), and anhydrous methanol (60 mL) were added to a reaction flask, and stirred under a hydrogen atmosphere at room temperature overnight. After the reaction was completed, the mother liquor was collected by suction filtration and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 4.3 g of the title compound as a brown-yellow oily liquid, with a yield of 74.1%.

[0477] LCMS: m / z 178.0 [M+H] + .

[0478] Step 3: Preparation of 3-iodo-7-isopropylthieno[3,2-b]pyridine (14c)

[0479] At room temperature, 7-isopropylthieno[3,2-b]pyridine (14b) (2.50 g, 14.1 mmol) and trifluoromethanesulfonic acid (25 mL) were added to the reaction flask, N-iodosuccinimide (2.40 g, 14.1 mmol) was added in batches at 0°C, the temperature was slowly restored to room temperature, and the reaction was allowed to proceed overnight. After the reaction was completed, the pH was adjusted to 8 with saturated sodium bicarbonate, ethyl acetate (100 mL) and water (50 mL) were added for extraction, the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 1.20 g of the title compound as a dark brown oily liquid, with a yield of 28.5%.

[0480] LCMS: m / z 303.90 [M+H] + .

[0481] Step 4: Preparation of 7-isopropyl-3-(trifluoromethyl)thieno[3,2-b]pyridine (14d)

[0482] At room temperature, 3-iodo-7-isopropylthieno[3,2-b]pyridine (14c) (1.10 g, 3.60 mmol), methyl fluorosulfonyldifluoroacetate (3.50 g, 18.1 mmol), cuprous iodide (3.80 g, 20.0 mmol), and N,N-dimethylformamide (10 mL) were added to a reaction flask and reacted overnight at 100°C in a nitrogen atmosphere. After cooling to room temperature and the reaction was completed, the mother liquor was collected by suction filtration and extracted with ethyl acetate (300 mL) and water (300 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 540 mg of the title compound as a yellow-brown oily liquid with a yield of 60.7%.

[0483] LCMS: m / z 246.0 [M+H] + .

[0484] Step 5: Preparation of 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-isopropyl-3-(trifluoromethyl)thieno[3,2-b]pyridine (14e)

[0485] At room temperature, 7-isopropyl-3-(trifluoromethyl)thieno[3,2-b]pyridine (14d) (245 mg, 1.00 mmol) and tetrahydrofuran (2.0 mL) were added to the reaction bottle. Under nitrogen atmosphere, a tetrahydrofuran solution of lithium diisopropylamide (2N, 3.5 mL, 7.00 mmol) was added dropwise at -78°C. After reacting at this temperature for 10 minutes, a tetrahydrofuran solution of 2-chloro-5-fluoropyrimidine (264 mg, 2.00 mmol) (1.0 mL) was added dropwise. After reacting at this temperature for 20 minutes, the mixture was slowly returned to room temperature and stirred overnight. After the reaction was completed, ethyl acetate (50 mL) and water (50 mL) were added for extraction. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 4:1) to obtain 150 mg of the title compound as a light yellow oily liquid with a yield of 40.0%.

[0486] LCMS: m / z 375.9 [M+H] + .

[0487] Step 6: Preparation of (3S,4R)-4-((5-fluoro-4-(7-isopropyl-3-(trifluoromethyl)thieno[3,2-b]pyridin-2-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (14)

[0488] At room temperature, 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-isopropyl-3-(trifluoromethyl)thieno[3,2-b]pyridine (14e) (100 mg, 0.26 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (37.4 mg, 0.32 mmol), N,N-diisopropylethylamine (68.7 mg, 0.53 mmol), N-methylpyrrolidone (NMP) (1 mL) were added to a reaction bottle and reacted at 130°C for 2 hours under microwave. After cooling to room temperature, ethyl acetate (20 mL) and water (5 mL) were added. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 15.0 mg of the off-white solid title compound, with a yield of 12.3%

[0489] LCMS: m / z 457.0 [M+H] + .

[0490] 1 H NMR (400MHz, DMSO) δ8.83(d,J=4.8Hz,1H),8.59(d,J=1.7Hz,1H),7.56(d,J=5.0Hz,2H),4.93(d,J=5.3Hz,1H),3.84-3.73(m,3H),3.54-3. 46(m,1H),3.35(s,1H),3.23(dd,J=13.7,6.8Hz,1H),3.07-2.99(m,1H),1.96(d,J=10.0Hz,1H),1.56-1.43(m,1H),1.39(d,J=6.8Hz,6H).

[0491] Example 15: Preparation of (3S,4R)-4-((5-fluoro-4-(7-isopropyl-3-methylthieno[3,2-c]pyridin-2-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (15)

[0492]

[0493] Step 1: Preparation of 2-bromo-4-chloro-7-isopropyl-3-methylthieno[3,2-c]pyridine (15a).

[0494] 2-Bromo-7-isopropyl-3-methylthieno[3,2-c]pyridin-4(5H)-one (6d) (500 mg, 1.75 mmol) was dissolved in phosphorus oxychloride (2.5 mL) at room temperature, and the reaction solution was stirred at 100°C for 2 h. After monitoring the completion of the reaction, the reaction solution was poured into ice water (10 mL) and stirred for 10 min. Ethyl acetate (15 mL) was added and allowed to stand for stratification. The aqueous phase was extracted with ethyl acetate (10 mL x 2), and the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain 520 mg of the title compound as a yellow solid, with a yield of 97%.

[0495] LCMS: m / z 303.9 [M+H] + .

[0496] Step 2: Preparation of 7-isopropyl-3-methylthieno[3,2-c]pyridine (15b).

[0497] At room temperature, 2-bromo-4-chloro-7-isopropyl-3-methylthieno[3,2-c]pyridine (15a) (430 mg, 1.42 mmol) was dissolved in methanol (5 mL), palladium / carbon (86 mg) was added, and the reaction solution was replaced with hydrogen gas three times and stirred at room temperature for 16 h. After monitoring the reaction completion, the reaction solution was filtered with diatomaceous earth and concentrated to obtain 260 mg of the title compound as a yellow solid with a yield of 96%. It can be used directly in the next step.

[0498] LCMS: m / z 192.10 [M+H] + .

[0499] Step 3: Preparation of 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-isopropyl-3-methylthieno[3,2-c]pyridine (15c).

[0500] 7-Isopropyl-3-methylthieno[3,2-c]pyridine (15b) (100 mg, 0.524 mmol) was dissolved in tetrahydrofuran (5 mL) at -78°C, and lithium diisopropylamide (1.83 mL, 3.67 mmol) was slowly added dropwise, and the reaction solution was stirred at -78°C for 10 min. 2-Chloro-5-fluoropyrimidine (138 mg, 1.05 mmol) was then added, and the reaction solution was stirred at -78°C for 18 h. Water (10 mL) was added to the reaction solution, and it was extracted with ethyl acetate (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain 50 mg of the title compound as a yellow solid, with a yield of 30%.

[0501] LCMS: m / z 321.90 [M+H] + .

[0502] Step 4: Preparation of (3S,4R)-4-((5-fluoro-4-(7-isopropyl-3-methylthieno[3,2-c]pyridin-2-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (15).

[0503] 2-(2-Chloro-5-fluoropyrimidin-4-yl)-7-isopropyl-3-methylthieno[3,2-c]pyridine (15c) (50 mg, 0.156 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (23.9 mg, 0.156 mmol) and diisopropylethylamine (40.2 mg, 0.312 mmol) were dissolved in N-methylpyrrolidone (3 mL) at room temperature, and the reaction solution was stirred at 120 °C for 18 h. After monitoring the completion of the reaction, the reaction solution was diluted with ethyl acetate (10 mL), washed with water (10 mL x 3), washed with saturated brine (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 11.0 mg of the title compound as a yellow solid, with a yield of 18%.

[0504] LC-MS: m / z 403.00 [M+H] + .

[0505] 1 H NMR (400MHz, DMSO) δ9.06 (s, 1H), 8.52 (d, J = 2.7Hz, 1H), 8.46 (s, 1H), 7.35 (d, J=7.8Hz,1H),4.92(d,J=5.2Hz,1H),3.82(dd,J=11.0,5.0Hz,3H),3.57-3.46 (m,1H),3.35(s,1H),3.20(dd,J=13.9,6.9Hz,1H),3.07-2.99(m,1H),2.62(d ,J=2.1Hz,3H),1.97(d,J=11.8Hz,1H),1.55-1.45(m,1H),1.45-1.32(m,6H).

[0506] Example 16: Preparation of 2-(5-fluoro-2-((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-7-isopropyl-3-methylthieno[3.2c]pyridin-4(5H)-one (16)

[0507]

[0508] Step 1: Preparation of 2-bromo-7-isopropyl-5-(4-methoxybenzyl)-3-methylthieno[3,2-c]pyridin-4(5H)-one (16a).

[0509] At room temperature, 2-bromo-7-isopropyl-3-methylthieno[3,2-c]pyridin-4(5H)-one (500 mg, 1.74 mmol) and cesium carbonate (1.13 g, 3.47 mmol) were dissolved in N,N-dimethylformamide (5 mL), p-methoxybenzyl bromide (523 mg, 2.60 mmol) was added dropwise, and the reaction solution was stirred at room temperature for 2 h. After monitoring the reaction completion, ethyl acetate (20 mL) was added to dilute the reaction solution, water (20 mL) was added, and the layers were separated by standing. The aqueous phase was extracted with ethyl acetate (20 mL x 2), and the combined organic phase was washed with saturated brine (20 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain 670 mg of the title compound as a yellow solid, with a yield of 95%.

[0510] LCMS: m / z 406.00 [M+H] + .

[0511] Step 2: Preparation of 7-isopropyl-5-(4-methoxybenzyl)-3-methyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thieno[3,2-c]pyridin-4(5H)-one (16b).

[0512] At room temperature, 2-bromo-7-isopropyl-5-(4-methoxybenzyl)-3-methylthieno[3,2-c]pyridin-4(5H)-one (600 mg, 1.48 mmol), bis(boronic acid pinacol) (452 ​​mg, 1.78 mmol), potassium acetate (290 mg, 2.96 mmol) and Pd(dppf)Cl were added. 2 (108 mg, 0.148 mmol) was dissolved in 1,4-dioxane (6 mL), and the reaction solution was replaced with gas three times under nitrogen protection and stirred at 100 ° C for 16 h. After monitoring the reaction completion, the reaction solution was diluted with ethyl acetate (20 mL), filtered and concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 4: 1) to obtain 140 mg of the title compound as a yellow solid, with a yield of 21%.

[0513] LCMS: m / z 454.00 [M+H] + .

[0514] Step 3: Preparation of 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-isopropyl-5-(4-methoxybenzyl)-3-methylthieno[3.2c]pyridin-4(5H)-one (16c).

[0515] 7-Isopropyl-5-(4-methoxybenzyl)-3-methyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thieno[3,2-c]pyridin-4(5H)-one (140 mg, 0.309 mmol), potassium phosphate (131 mg, 0.618 mmol) and 2,4-dichloro-5-fluoropyrimidine (77.5 mg, 0.464 mmol) were dissolved in a mixed solvent of 1,4-dioxane (4 mL) and water (1 mL) at room temperature, and Pd(dppf)Cl was added. 2 (22.6 mg, 0.0309 mmol), the reaction solution was replaced with gas three times under nitrogen protection, and stirred at 100 ° C for 16 h. After monitoring the reaction completion, ethyl acetate (20 mL) was added to dilute the reaction solution, water (20 mL) was added, and the layers were separated by standing. The aqueous phase was extracted with ethyl acetate (20 mL x 2), and the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain 120 mg of the title compound as a yellow solid, with a yield of 85%.

[0516] LCMS: m / z 459.00 [M+H] + .

[0517] Step 4: Preparation of 2-(5-fluoro-2-((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-7-isopropyl-5-(4-methoxybenzyl)-3-methylthieno[3.2c]pyridin-4(5H)-one (16d).

[0518] At room temperature, 2-(2-chloro-5-fluoropyrimidin-4-yl)-7-isopropyl-5-(4-methoxybenzyl)-3-methylthieno[3.2c]pyridin-4(5H)-one (120 mg, 0.262 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (40.1 mg, 0.262 mmol) and diisopropylethylamine (67.6 mg, 0.524 mmol) were dissolved in N-methylpyrrolidone (3 mL), and the reaction solution was stirred at 120 °C for 16 h. After monitoring the completion of the reaction, the reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 mL x 3), washed with saturated brine (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 1: 1) to give 75 mg of the title compound as a yellow oil with a yield of 53%.

[0519] LCMS: m / z 539.00 [M+H] + .

[0520] Step 5: Preparation of 2-(5-fluoro-2-((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-7-isopropyl-3-methylthieno[3.2c]pyridin-4(5H)-one (16).

[0521] 2-(5-Fluoro-2-((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-7-isopropyl-5-(4-methoxybenzyl)-3-methylthieno[3.2c]pyridin-4(5H)-one (75 mg, 0.139 mmol) was dissolved in trifluoroacetic acid (3 mL) at room temperature, and the reaction solution was stirred at 120 °C for 18 h. After monitoring the completion of the reaction, the reaction solution was concentrated, diluted with ethyl acetate (20 mL), washed with saturated sodium bicarbonate aqueous solution (10 mL x 3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to give 17.5 mg of the title compound as a yellow solid, with a yield of 30%.

[0522] LC-MS: m / z 419.00 [M+H] + .

[0523] 1 H NMR (400MHz, DMSO) δ11.38(s,1H),8.45(s,1H),7.25(d,J=7.8Hz,1H),7.07(s,1H),4.90(d,J=4.6Hz,1H),3.80(d,J=5.2Hz,3H),3.50(s,1H) ,3.35(s,1H),3.03(t,J=10.5Hz,1H),2.93-2.81(m,1H),2.67(s,3H),1.96(d,J=10.4Hz,1H),1.47(d,J=10.4Hz,1H),1.28(d,J=5.6Hz,6H).

[0524] Example 17: Preparation of 2-(2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)-5-methylpyrimidin-4-yl)-7-isopropylthieno[3,2-c]pyridin-4(5H)-one (17)

[0525]

[0526] Step 1: Preparation of (7-isopropyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridin-2-yl)boronic acid (17a)

[0527] At room temperature, 2-bromo-7-isopropylthieno[3,2-c]pyridin-4(5H)-one (10d) (1.0 g, 3.69 mmol), bis(boronic acid pinacol) (1.4 g, 5.53 mmol), Pd(dppf)Cl 2 (270.4 mg, 0.37 mmol), potassium acetate (1.0 g, 11.07 mmol), and 1,4-dioxane (10 mL) were added to the reaction flask, and the reaction was stirred at 100 ° C in a nitrogen atmosphere overnight. After cooling to room temperature, ethyl acetate (50 mL) and water (50 mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: dichloromethane: methanol = 5:1) to obtain 800 mg of the title compound as a black solid, with a yield of 91.5%.

[0528] LCMS: m / z 238.1 [M+H] + .

[0529] Step 2: Preparation of 2-(2-chloro-5-methylpyrimidin-4-yl)-7-isopropylthieno[3,2-c]pyridin-4(5H)-one (17b)

[0530] At room temperature, (7-isopropyl-4-oxo-4,5-dihydrothieno[3,2-c]pyridin-2-yl)boronic acid (17a) (800.0 mg, 3.37 mmol), 2,4-dichloro-5-methylpyrimidine (820 mg, 5.05 mmol), Pd(dppf)Cl 2 (246.4 mg, 0.33 mmol), potassium phosphate (2.1 g, 10.12 mmol), 1,4-dioxane (10 mL), and water (1 mL) were added to the reaction flask, and the reaction was stirred at 100 ° C in a nitrogen atmosphere overnight. After cooling to room temperature, ethyl acetate (50 mL) and water (50 mL) were added for extraction. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: dichloromethane: methanol = 5:1) to obtain 600 mg of the title compound as a black solid, with a yield of 55.7%.

[0531] LCMS: m / z 320.0 [M+H] + .

[0532] Step 3: Preparation of 2-(2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)-5-methylpyrimidin-4-yl)-7-isopropylthieno[3,2-c]pyridin-4(5H)-one (17)

[0533] At room temperature, 2-(2-chloro-5-methylpyrimidin-4-yl)-7-isopropylthieno[3,2-c]pyridin-4(5H)-one (17b) (100 mg, 0.30 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol hydrochloride (71.9 mg, 0.47 mmol), diisopropylethylamine (121.1 mg, 0.94 mmol), and N-methylpyrrolidone (1 mL) were added to the reaction bottle and stirred at 120°C overnight. After cooling to room temperature, ethyl acetate (20 mL) and water (5 mL) were added. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 43.0 mg of the light yellow solid title compound, with a yield of 34.4%.

[0534] LCMS: m / z 401.1 [M+H] + .

[0535] 1 H NMR (400MHz, DMSO) δ8.03(d,J=0.8Hz,1H),7.71(d,J=5.3Hz,1H),7.56(d,J=5. 3Hz,1H),7.31(s,1H),6.87(d,J=7.7Hz,1H),4.89(d,J=5.5Hz,1H),4.04-3.87 (m,1H),3.87-3.71(m,2H),3.70-3.54(m,1H),3.29-3.23(m,1H),3.08-2.83(m ,2H),2.10(s,3H),1.98-1.90(m,1H),1.61-1.46(m,1H),1.29(d,J=8.0Hz,6H).

[0536] Biological tests

[0537] Experimental Example 1: In vitro kinase assay

[0538] The ADP-Glo ​​assay (Promega, V9102) was used to detect the inhibitory activity of the test compound on CDK4 / CyclinD1 and CDK6 / CyclinD3 kinases.

[0539] First, the compound was dissolved in DMSO (Sigma, D8418), the experimental starting concentration was 10 μM, 3-fold dilution, 10 gradients. 5 μL of the diluted compound and 2.5 μL of kinase (CDK4 / Cyclin D1: Invitrogen, PV4436; CDK6 / CyclinD3: Carna, 04-107) were added to a 384-well reaction plate (Greiner, 784075) using Echo 550, the plate was sealed with a sealing film, centrifuged at 1000 rpm for 1 min, and incubated at 25°C for 10 min. Then 2.5 μL of a mixture of substrate Histone H1 protein (SignalChem, H10-54N) and ATP (Promega, V910B) was added to the reaction plate, centrifuged at 1000g for 30 s, and incubated at room temperature for 2 h. 4 μL of ADP-GLO reagent (Promega, V9102) was pipetted into the reaction plate, centrifuged briefly, and incubated at room temperature for 40 min. Finally, 10 μL of detection reagent was added to the reaction plate and incubated for 40 min. The relative luminescence unit (RLU) value was read using the Envision 2104 multi-function plate reader (PerkinElmer, Oct-04). The RLU value is used to characterize the degree of reaction between the enzyme and the substrate.

[0540] The experimental data were calculated using GraphPad prism 8.0 software with nonlinear fitting formula.

[0541] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope)).

[0542] X is the log value of compound concentration; Y is the inhibition level of kinase; Top and Bottom are the Y values ​​of the highest and lowest plateaus of the curve; Hillslope is the Hill constant.

[0543] The inhibitory activity of the compounds of the present invention on various kinases is shown in Table 1 below, where: A represents IC 50 Value is less than 10nM, B represents IC 50 The value is 10-100nM, and C represents IC 50 The value is 100-500nM, and D represents IC 50 The value is greater than 500nM.

[0544] Table 1 Kinase inhibitory activity of the compounds of the present invention

[0545]

[0546] Conclusion: It can be seen from Table 1 that the compounds of the present invention have good CDK4 kinase inhibitory activity.

Claims

1. A compound represented by the general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, in, Ring A is a 5-6 membered heteroaryl group; X is CR 7 or N; Y is CR 8 or N; Z is CR 9 or N; Q is NR 10 , O or CR 11 R 12 , where R 11 and R 12 Together with the C atoms to which they are attached, they form a 13 or a 4-6 membered heterocyclic ring with O as a ring member; Each R 3 independently selected from hydrogen, deuterium, halogen, oxo, hydroxyl, cyano, nitro, alkyl, alkoxy, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, alkylthioether, -NR a R b 、-C(O)R c 、-OC(O)R c 、-C(O)OR c 、-C(O)NR a R b 、-S(O) p R c 、-S(O) p NR a R b 、-S(O)(=NR a )R c 、-SF5、-P(O)R a R b 、-P(O)(OR c )(OR d ) and -B(OR c )(OR d ), wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted by one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R 4 Selected from hydrogen atom, halogen, -NR a R b , nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R 5 Selected from hydrogen atom, halogen, -NR a R b , nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R 6 Selected from hydrogen atom, halogen, -NR a R b , nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R 7 , R 8 and R 9 are each independently selected from hydrogen, halogen, -NR a R b , nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R 10 and R 13 are independently selected from hydrogen, alkyl, halogenated alkyl, -S(O) p R c 、-S(O) p NR a R b 、-C(O)R a 、-C(O)OR a and -C(O)NR a R b , the alkyl and haloalkyl are each independently optionally further selected from halogen, -NR a R b , nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroaryl, -S(O) p R c 、-S(O) p NR a R b 、-C(O)R a 、-C(O)OR a 、-C(O)NR a R b and -OC(O)R a is substituted by one or more groups; R a , R b , R c and R d each independently selected from hydrogen, halogen, hydroxy, cyano, amino, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxy, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; Or, R a and R b Together with the nitrogen atom to which they are attached, they form a heterocyclic group, which is optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl; p is 1 or 2; q is 1, 2, 3 or 4.

2. The compound of the general formula (I) according to claim 1, or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, which is the compound of the general formula (II) or its stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, in, Ring A, R 3 , R 4 , R 5 , R 6 , R 7 , Q and q are as defined in claim 1.

3. A compound of the general formula (I) according to claim 1 or 2, or a stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, which is a compound of the general formula (III) or general formula (IV), or a stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, in, Ring A, R 3 , R 4 , R 5 , R 6 , R 7 , R 10 and q as defined in claim 1.

4. A compound of the general formula (I) according to any one of claims 1 to 3, or a stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, which is a compound of the general formula (V) or general formula (VI), or a stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, in, Ring A, R 3 , R 4 , R 5 , R 7 and q as defined in claim 1.

5. A compound of the general formula (I) according to any one of claims 1 to 4, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Ring A is selected from pyrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, triazolyl, tetrazolyl, pyrrolyl, furanyl, thienyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl and triazinyl, preferably pyrazolyl, imidazolyl and pyridinyl.

6. A compound of the general formula (I) according to any one of claims 1 to 5, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Each R 3 independently selected from hydrogen atoms, deuterium atoms, halogens, oxo groups, hydroxyl groups, cyano groups, nitro groups, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl, preferably hydrogen, halogen, oxo, hydroxyl, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-6 Cycloalkyl; and / or q is 1 or 2.

7. A compound of the general formula (I) according to any one of claims 1 to 6, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Selected from 8. A compound of the general formula (I) according to any one of claims 1 to 7, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 4 Selected from hydrogen, halogen, -NH2, nitro, cyano, hydroxyl, mercapto, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl, preferably hydrogen, halogen, -NH2, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 Hydroxyalkyl, more preferably hydrogen atom, halogen, C 1-6 Alkyl and C 1-6 Haloalkyl; and / or R 5 Selected from hydrogen, halogen, -NH2, nitro, cyano, hydroxyl, mercapto, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl, preferably hydrogen, halogen, -NH2, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 Hydroxyalkyl, more preferably halogen and C 1-6 alkyl; and / or R 7 Selected from hydrogen, halogen, -NH2, nitro, cyano, hydroxyl, mercapto, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl, preferably hydrogen, halogen, -NH2, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 A hydroxyalkyl group is more preferably a hydrogen atom.

9. A compound of the general formula (I) according to any one of claims 1 to 3 and 5 to 8, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 6 Selected from hydrogen, halogen, -NH2, nitro, cyano, hydroxyl, mercapto, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl, preferably hydrogen, halogen, -NH2, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 A hydroxyalkyl group is more preferably a hydroxy group.

10. A compound of the general formula (I) according to any one of claims 1 to 9, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R a and R b are each independently selected from hydrogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, 3-6 membered heterocyclic group, wherein the C 1-6 Alkyl, C 3-8 The cycloalkyl and 3-6 membered heterocyclic groups are each independently further selected from halogen, amino, cyano, hydroxyl, mercapto, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 One or more groups in the haloalkoxy group are substituted; Or, R a and R b Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group is optionally further selected from halogen, amino, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl and C 1-6 One or more groups in the haloalkoxy group are substituted.

11. A compound of the general formula (I) according to any one of claims 1 to 10, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

12. A method for preparing a compound of the general formula (I) according to any one of claims 1 to 11 or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, comprising the following steps: In a solvent, in the presence of a base, compound Id reacts with compound Ie to obtain a compound represented by general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof; The solvent is, for example, N-methylpyrrolidone, and the base is, for example, N,N-diisopropylethylamine; Among them, rings A, X, Y, Z, Q, R 3 , R 4 , R 5 , R 6 and q as defined in claim 1.

13. A pharmaceutical composition comprising a compound of the general formula (I) according to any one of claims 1 to 11 or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

14. Use of a compound of the general formula (I) according to any one of claims 1 to 11 or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 13 in the preparation of a cyclin-dependent kinase (CDK) inhibitor.

15. Use of the compound represented by the general formula (I) according to any one of claims 1 to 11 or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereoisomer, or mixture thereof, or its pharmaceutically acceptable salt, or the pharmaceutical composition according to claim 13 in the preparation of a drug for inhibiting cancer cell proliferation, inhibiting cancer cell invasion, or inducing cancer cell apoptosis.

16. Use of a compound of formula (I) according to any one of claims 1 to 11 or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 13 in the preparation of a medicament for preventing and / or treating a disease associated with cyclin-dependent kinase (CDK) activity, such as cancer, in particular a cancer characterized by amplification or overexpression of CDK4 and cyclin D3, preferably breast cancer.

17. The use according to any one of claims 15 to 16, wherein the compound represented by the general formula (I) according to any one of claims 1 to 10 or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereoisomer, or mixture thereof, or its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 13 can be administered or used in combination with another anticancer therapeutic agent or anticancer treatment method simultaneously, separately or sequentially.

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