WO2022253328A1

WO2022253328A1 - Compound having kinase inhibitory function, and preparation method therefor and application thereof

Info

Publication number: WO2022253328A1
Application number: PCT/CN2022/096950
Authority: WO
Inventors: 江磊; 程战领; 冯志勇; 金贤; 刘胜洋; 聂士常; 石倩; 寿建勇; 汪兵; 许灵灵; 徐圆; 张建华; 张毅; 赵海霞
Original assignee: 轶诺(浙江)药业有限公司; 上海轶诺药业有限公司
Priority date: 2021-06-04
Filing date: 2022-06-02
Publication date: 2022-12-08
Also published as: CN115433161A; CN117751104A

Abstract

An HPK1 kinase inhibitor, and a preparation method therefor and an application thereof. Specifically, a compound shown in formula (I), the compound having HPK1 inhibitory activity and being used for preparing a pharmaceutical composition for treating cancer and other diseases related to HPK1 activity.

Description

A class of compounds with kinase inhibitory function and their preparation and application

technical field

The invention relates to the field of small molecule drugs, in particular, the invention relates to a kinase inhibitor and its preparation and application.

Background technique

T cells, B cells and dendritic cells (DCs) are powerful weapons that the body's immune system uses to fight against foreign invaders such as viruses and bacteria, as well as self-infected or abnormal cells such as cancerous cells. The human body has a complex and sophisticated regulatory system to ensure the normal operation of the immune system. When cancerous cells appear in the human body, if the immune system cannot completely kill them, these immune-evading cancer cells proliferate abnormally and form tumors. Traditional tumor treatment is mainly through surgery, radiotherapy, chemotherapy and molecular targeted drugs. However, surgical resection is often not a viable option for many forms of tumor or cancer. While radiation therapy and chemotherapy target tumor cells, they also damage some healthy cells. Tumor cells are prone to mutations and can develop resistance to drugs that specifically target tumor cells, making cancer treatment difficult. In recent years, using the patient's own immune system to overcome the immune escape strategy adopted by tumor cells and enhance the body's anti-tumor immunity is a new cancer treatment strategy. One such strategy is to overcome tumor cell immune escape by inhibiting negative regulators of the immune response that normally function to maintain peripheral tolerance, allowing tumor antigens to be recognized as nonself antigens. Hematopoietic progenitor kinase 1 (HPK1), also known as MAP4K1 (a member of the MAP4K family), is a negative regulator of the activation of dendritic cells (DCs), T cells, and B cells. Inhibiting its activity can specifically enhance the body's anti-tumor immunity. HPK1 is predominantly expressed by hematopoietic cells, including early hematopoietic progenitor cells. In T cells, HPK1 is considered to be able to degrade these proteins by phosphorylating the Ser376 site of the downstream SLP76 protein and the Thr254 site of the Gads protein and recruiting the 14-3-3 protein, thereby reducing the persistence of signal microclusters and playing a role to negatively regulate T cell activation. HPK1 can also be activated in response to prostaglandins (PGE2), which are normally secreted by tumors, helping tumor cells escape from the immune system. Moreover, HPK1 can also inhibit AP-1, and AP-1 plays a role in promoting cell proliferation, inhibiting differentiation, and promoting tumor cell invasion and metastasis in the process of tumor formation and development. Targeted disruption of alleles of HPK1 kinase allows T cells to increase Th1 cytokine (IL-2, IFNγ, etc.) production in TCR responses. HPK1 has multiple roles in immunity and has been implicated in the pathogenesis of autoimmune diseases, cancer and inflammatory responses. HPK1 kinase knockout (HPK1-/-) T cells proliferate much faster than monomeric wild type, and mice transfected with HPK1-/- T cells can resist tumor growth, and dendritic cells (DCs) that lose HPK1 kinase ) has better antigen presentation ability than wild type, and can better exhibit anti-tumor immune response. In addition, animal experiments have shown that HPK1 inhibition and PD-1/PD-L1 antibody drugs have obvious synergistic anti-tumor activity. Therefore, HPK1 kinase has a key role in disease treatment, especially cancer treatment.

At present, there is no drug on the market for this target. In order to meet the huge clinical needs in the future, we hope to design and develop a selective and highly active small molecule HPK1 inhibitor to provide a new oral drug for the treatment of immune-related diseases, especially tumors. Drugs, used alone or in combination with chemotherapy, radiotherapy, tumor-targeted drugs, other tumor immunotherapeutic agents (small molecule compounds and antibodies), and tumor vaccines, etc.

Contents of the invention

The purpose of the present invention is to provide a small molecule HPK1 inhibitor with selectivity and high activity.

The first aspect of the present invention provides a compound represented by the following formula I, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof;

in,

- M ² , M ³ - each independently CR ¹ ;-

^R is selected from the group consisting of CN, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, phenyl, substituted or unsubstituted C3-C8 cycloalkyl, or _-CONH2 ;

A - selected from the following group:

Wherein, V ¹ , V ² , V ³ , V ⁴ , V ⁵ , V ⁶ , V ⁷ or V ⁸ are each independently CR ² or N or NR ² ;

Each R ² is independently selected from the group consisting of H, CN, halogen, NH ₂ , substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, phenyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group with 1-3 heteroatoms selected from N, S and O in the following group, -C(O)-(substituted or unsubstituted A 3-12-membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O), -(CH ₂ ) _m N(R ³ ) ₂ , -C(O)(CH ₂ ) _m N(R ³ ) ₂ , -NHC(O)(CH ₂ ) _m N(R ³ ) ₂ ; wherein each of said R ³ is independently selected from the following group: H, substituted or unsubstituted C1-C6 Alkyl group, or two R ³ together with the connected N atoms form a 4-7 membered nitrogen-containing heterocyclic ring;

Or two ^R2 located on adjacent ring atoms together form a 5-7 membered substituted or unsubstituted cyclic group;

m is 0, 1, 2, 3, 4 or 5;

Ra has the structure shown in the following formula:

Wherein, the ring B is a substituted or unsubstituted C3-C8 cycloalkyl group, a substituted or unsubstituted 6-10 membered aryl group, a substituted or unsubstituted 5-12 membered heteroaryl group, a substituted or unsubstituted A 3-15 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O; and the ring B is a bicyclic structure;

n is 0, 1, 2, 3, 4 or 5;

^Each R is independently selected from the group consisting of H, halogen, CN, oxygen atom (=O), substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted Substituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O, substituted or unsubstituted C6-C10 aryl , a substituted or unsubstituted 3-10 membered heteroaryl group having 1-3 heteroatoms selected from the group consisting of N, S and O, R ⁷ -C(=O)-, R ⁷ -C(=O) -3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O-, R ⁸ -S(=O) ₂ -, R ⁹ R ¹⁰ NC(=O)-, R ⁹ R ¹⁰ NC(=O)NR-, R ¹¹ R ¹² NS(=O) ₂ -, R ⁷ O-, R ⁸ -S(O) ₂ NR-; and two R ⁴ can together form a substituted or unsubstituted 5-7 membered carbocyclic or heterocyclic ring (including saturated or unsaturated ring);

Each R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , and R are each independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkane Oxygen, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 6-10 membered aryl, substituted or unsubstituted with 1-3 heteroatoms selected from the group consisting of N, S and O 3-12 membered heterocyclic group; or said R ⁹ and R ¹⁰ or R ¹¹ and R ¹² together form a substituted or unsubstituted nitrogen atom with 1-3 heteroatoms selected from N, S and O The 3-12 membered heterocyclic group;

Unless otherwise specified, the "substitution" refers to being substituted by one or more (such as 2, 3, 4, etc.) substituents selected from the following group: D, halogen, hydroxyl, C1-C6 alkoxy Group, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, methylsulfone, oxo (=O), -CN, -NH ₂ , -NHS (O) ₂ CH ₃ , C1-C6 amino group, carboxyl group, C1-C6 amido group (-C(=O)-N(Rd) ₂ or -NH-C(=O)(Rd), Rd is H or C1-C5 alkyl), or unsubstituted or substituted by one or more substituents selected from Z group, selected from the following groups: C1-C6 alkyl, C6-C10 aryl, with 1-3 5-10 membered heteroaryl, -(CH ₂ )-C6-C10 aryl, -(CH ₂ )-(having 1-3 heteroatoms selected from N, S and O 3-10 membered heteroaryl group with heteroatoms), -(5-10 membered heteroarylene group with 1-3 heteroatoms selected from N, S and O)-(C1-C6 alkyl), with 1-3 3-12-membered heterocyclic groups (including monocyclic, spiro, bridged or parallel rings) with 1-3 heteroatoms selected from the following group of N, S and O, and the group Z substituents are selected from the following Groups: halogen, C1-C6 alkyl, C1-C6 alkylene-OH, C1-C6 alkoxy, oxo, -S(O) ₂ CH ₃ , -CN, -OH, C6-C10 aryl, A 3-10 membered heterocyclic group having 1-3 heteroatoms selected from N, S and O, -C(O)CH ₂ NH ₂ , -C(O)CH ₂ OH;

And in the compound of formula I, each chiral center is in R configuration or S configuration.

In another preferred example, the ring A has at least one substituent.

In another preferred example, the ring B is selected from the following group: 5-6 membered aryl (5-7 membered partially unsaturated or aromatic heterocycle), 5-7 membered heteroaryl (5 -7-membered partially unsaturated or aromatic heterocycle), 3-10-membered heterocyclic group with 1-3 heteroatoms selected from the group consisting of N, S and O (5-7-membered partially unsaturated or aromatic Heterocycle), [3-8 membered saturated or partially unsaturated ring] spiro [3-8 membered saturated or partially unsaturated ring] (can be carbocyclic or heterocyclic), or at least one selected from the following group Substituent substituted 6-7 membered aryl or 5-7 membered heteroaryl: substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 6-10 membered aryl, substituted or unsubstituted with 1 - a 3-12 membered heterocyclic group having 3 heteroatoms selected from the group consisting of N, S and O.

In another preferred example, the ring B is a substituted or unsubstituted group selected from the following group:

Wherein, said U ¹ , U ² , U ³ , U ⁴ , U ⁵ , U ⁶ , U ⁷ , and U ⁸ are each independently selected from the following group: CR ¹ or N;

Said Y ¹ , Y ² , Y ³ , Y ⁴ or Y ⁵ are each independently selected from the following group: None, or C(R ⁵ ) ₂ , C(R ⁵ ) ₂ C(R ⁵ ) ₂ , NR ⁵ , O or S;

Each of J and K is independently selected from the group consisting of none, or C(R ⁶ ) ₂ , NR ⁶ , O or S;

Wherein, p is 0, 1, 2 or 3;

q is 0, 1, 2 or 3;

Each R ⁵ is independently selected from the group consisting of H, CN, halogen, NH ₂ , substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, phenyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group (preferably 3-6 membered heterocyclic group) having 1-3 heteroatoms selected from N, S and O in the following group, -(CH ₂ ) _m N(R ³ ) ₂ , -C(O)N(R ³ ) ₂ , -NHC(O)(CH ₂ ) _m N(R ³ ) ₂ ; wherein, each of the R ³ Independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, or two R ³ together with the connected N atoms form a 4-7 membered nitrogen-containing heterocyclic ring;

Or two R ⁵ jointly constitute a group selected from the group consisting of: =O, -(J) _n -;

Or ^R5 located on two adjacent ring atoms and the connected ring atoms together form a substituted or unsubstituted 5-7 membered partially unsaturated or aromatic heterocyclic ring;

Each R ⁶ is independently selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl;

The linking site can be located on any ring atom of U ¹ , U ² , U ³ , U ⁴ , U ⁵ , U ⁶ , U ⁷ , U ⁸ , Y ¹ , Y ² , Y ³ , Y ⁴ , J and K .

In another preferred example, the ring B is a substituted or unsubstituted group selected from the group consisting of 6-7 membered aryl (5-7 membered partially unsaturated or aromatic heterocycle), 5- 7-membered heteroaryl (5-7 membered partially unsaturated or aromatic heterocycle).

In another preferred example, said M ³ is CR _b ; and said R _b is selected from the group consisting of CN, halogen, -C(O)N(R ⁶ ) ₂ , substituted or unsubstituted C1- C6 alkyl.

In another preferred embodiment, ring A is selected from the following group:

Wherein, V ¹ , V ² , V ³ , V ⁴ , V ⁵ , V ⁶ or V ⁷ are each independently CR ² or N; V ⁸ is NR ² ; each R ² is independently selected from the following group: H, CN, halogen, NH ₂ , substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, phenyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted A 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group N, S and O, -C(O)-substituted or unsubstituted having 1-3 heteroatoms selected from the group N, S and O 3-12 membered heterocyclic group with O heteroatom, -(CH ₂ ) _m N(R ³ ) ₂ , -C(O)(CH ₂ ) _m N(R ³ ) ₂ , -NHC(O)(CH ₂ ) _m N(R ³ ) ₂ ; wherein, each of said R ³ is independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, or two R ³ together with the connected N atom 4-7 membered nitrogen-containing heterocyclic ring.

In another preferred example, the A ring is selected from the following group:

Wherein, said R ² are each independently selected from the following group: H, CN, halogen, unsubstituted or halogenated C1-C4 alkyl, unsubstituted or halogenated C1-C4 alkoxy;

And said R _c is selected from the following group: substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered with 1-3 heteroatoms selected from the group N, S and O Heterocyclic group, -C(O)-substituted or unsubstituted 3-12 membered heterocyclic group with 1-3 heteroatoms selected from the group consisting of N, S and O, -(CH ₂ ) _m N(R ³ ) ₂ , -C(O)(CH ₂ ) _m N(R ³ ) ₂ , -NHC(O)(CH ₂ ) _m N(R ³ ) ₂ ; wherein, the R ³ are each independently selected from The following group: H, substituted or unsubstituted C1-C6 alkyl, or two R ³ together with the connected N atoms form a 4-7 membered nitrogen-containing heterocyclic ring.

In another preferred example, the compound of formula I has the structure shown in the following formula II-1 or II-2:

in,

The R _b are each independently selected from the group consisting of halogen, CN, substituted or unsubstituted C1-C6 alkyl.

In another preferred embodiment, the compound of formula I is selected from the following group:

The second aspect of the present invention provides a pharmaceutical composition comprising (1) the compound as described in the first aspect of the present invention or its stereoisomer or tautomer, or A pharmaceutically acceptable salt, hydrate or solvate; (2) a pharmaceutically acceptable carrier.

The third aspect of the present invention provides the compound as described in the first aspect of the present invention or its stereoisomer or tautomer, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or as described herein The use of the pharmaceutical composition described in the second aspect of the invention is used to prepare a pharmaceutical composition for preventing and/or treating diseases related to the activity or expression of HPK1 kinase.

The fourth aspect of the present invention provides a compound described in the first aspect of the present invention and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates and deuterated compounds and other tumor immunotherapy Combination tumor chemotherapy regimens with anti-tumor agents, the other anti-tumor immunotherapeutic agents are selected from the following group: small molecule compounds and antibodies (including but not limited to PD-1, PD-L1, CTLA-4, TIM-3, TGF-β and Its receptors, LAG3 antagonists or TLR4, TLR7, TLR8, TLR9, STING agonists, etc.), chemotherapy regimens, radiotherapy regimens, tumor targeting drugs, tumor vaccines, etc.

In another preferred example, the other tumor immunotherapeutic agent may be administered before, after or simultaneously with the agent, or may be co-administered with other known therapies. It can also be used as a vaccine adjuvant.

The fifth aspect of the present invention provides the compound as described in the first aspect of the present invention and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates and deuterated compounds and CAR-T immunotherapy Combination applications in cancer immunotherapy.

In another preferred example, the diseases include but not limited to cancer, metastatic cancer, inflammation and autoimmune related diseases.

In another preferred example, the diseases include but are not limited to: lymphoma, blastoma, medulloblastoma, retinoblastoma, sarcoma, liposarcoma, synovial cell sarcoma, neuroendocrine tumor, carcinoid tumor, Gastrinoma, islet cell carcinoma, mesothelioma, schwannoma, acoustic neuroma, meningioma, adenocarcinoma, melanoma, leukemia or lymphoid malignancy, squamous cell carcinoma, epithelial squamous cell carcinoma, lung cancer, small Lung cell lung cancer, non-small cell lung cancer, adenocarcinoma lung cancer, lung squamous cell carcinoma, peritoneal carcinoma, hepatocellular carcinoma, gastric cancer, intestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, breast cancer Carcinoma, metastatic breast cancer, colon cancer, rectal cancer, colorectal cancer, uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, anal cancer, penile cancer, Merkel cell cancer, esophageal cancer, Biliary tract neoplasms, head and neck cancers, hematologic malignancies, nasopharyngeal carcinoma, multiple myeloma, large field villous adenoma, non-Hodgkin's lymphoma, bone cancer, testicular cancer, Hodgkin's disease, seminoma, Oral cancer, brain cancer, skin cancer, breast ductal carcinoma, renal pelvis carcinoma, Wilms tumor, esophageal adenoma, retinoblastoma, glioma, neurofibroma, gastrointestinal stromal tumor, carcinoma in situ, uterus Endometrial cancer and myelodysplastic syndrome, etc.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, we will not repeat them here.

Detailed ways

After long-term and in-depth research, the inventors designed and synthesized a new class of HPK1 kinase inhibitors. On this basis, the inventors have completed the present invention.

the term

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, the term "about" when used in reference to a specifically recited value means that the value may vary by no more than 5% from the recited value. For example, as used herein, the expression "about 100" includes 95 and 105 and all values in between (eg, 95.1, 95.2, 95.3, 95.4, etc.).

As used herein, the term "comprises" or "includes (comprising)" can be open, semi-closed and closed. In other words, the term also includes "consisting essentially of", or "consisting of".

definition

As used herein, the term "alkyl" includes straight or branched chain alkyl groups. For example, C ₁ -C ₈ alkyl means straight or branched chain alkyl with 1-8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl Wait.

As used herein, the term "alkenyl" includes straight or branched chain alkenyl groups. For example, C ₂ -C ₆ alkenyl refers to a linear or branched alkenyl group with 2-6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2 -Butenyl, or similar groups.

As used herein, the term "alkynyl" includes straight or branched chain alkynyl groups. For example, C ₂ -C ₆ alkynyl refers to a straight or branched alkynyl group having 2-6 carbon atoms, such as ethynyl, propynyl, butynyl, or similar groups.

As used herein, the term "C ₃ -C ₈ cycloalkyl" refers to a cycloalkyl group having 3-8 carbon atoms. It may be a single ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or similar groups. Bicyclic forms such as bridged or spiro forms are also possible.

As used herein, the term "C ₁ -C ₈ alkoxy" refers to a straight-chain or branched alkoxy group having 1-8 carbon atoms; for example, methoxy, ethoxy, propoxy, iso Propoxy, butoxy, isobutoxy, tert-butoxy, etc.

As used herein, the term "3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O" means a heterocyclic group having 3-12 atoms and wherein 1-3 atoms are selected from A saturated or partially saturated cyclic group of heteroatoms from the group N, S and O. It may be monocyclic or polycyclic, such as bridged, spiro or fused rings. Specific examples may be oxetane, azetidine, tetrahydro-2H-pyranyl, piperidyl, tetrahydrofuranyl, morpholinyl, pyrrolidinyl and the like.

As used herein, the term "C ₆ -C ₁₀ aryl" refers to an aryl group having 6-10 carbon atoms, for example, phenyl or naphthyl and the like.

As used herein, the term "5-10 membered heteroaryl group having 1-3 heteroatoms selected from the group consisting of N, S and O" refers to a heteroaryl group having 5-10 atoms and wherein 1-3 atoms are selected from A cyclic aromatic group of heteroatoms of N, S and O in the following group. It may be a single ring or a condensed ring. Specific examples may be pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl and (1,2, 4)-triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, etc.

Unless otherwise specified, groups of the present invention may be substituted by substituents selected from the group consisting of halogen, nitrile, nitro, hydroxyl, amino, C ₁ -C ₆ alkyl-amino, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy, halogenated C ₁ -C ₆ alkyl, halogenated C ₂ -C ₆ alkenyl, halo Substituted C ₂ -C ₆ alkynyl, halogenated C ₁ -C ₆ alkoxy, allyl, benzyl, C ₆ -C ₁₂ aryl, C ₁ -C ₆ alkoxy-C ₁ -C ₆ alkane C ₁ -C ₆ alkoxy-carbonyl, phenoxycarbonyl, C ₂ -C ₆ alkynyl-carbonyl, C ₂ -C ₆ alkenyl-carbonyl, C ₃ -C ₆ cycloalkyl-carbonyl, C ₁ -C ₆ alkyl-sulfonyl and the like.

As used herein, "halogen" or "halogen atom" refers to F, Cl, Br, and I. More preferably, halogen or halogen atoms are selected from F, Cl and Br. "Halosubstituted" means substituted with an atom selected from F, Cl, Br, and I.

Unless otherwise specified, the structural formulas described in the present invention are intended to include all isomeric forms (such as enantiomers, diastereoisomers and geometric isomers (or configurational isomers)): for example, those containing The R, S configuration of the symmetrical center, the (Z), (E) isomer of the double bond, etc. Accordingly, individual stereochemical isomers of the compounds of the present invention or mixtures thereof as enantiomers, diastereomers or geometric isomers (or configurational isomers) are within the scope of the present invention.

As used herein, the term "tautomer" means that structural isomers with different energies can interconvert beyond a low energy barrier. For example, proton tautomers (ie, prototropism) include interconversions via migration of a proton, such as 1H-indazole and 2H-indazole. Valence tautomers include interconversion by recombination of some of the bonding electrons.

As used herein, the term "solvate" refers to a complex in which a compound of the present invention coordinates with solvent molecules to form specific ratios.

Compound of formula I

The invention provides a compound shown in the following formula I:

in,

X is selected from the group consisting of None, NR, S, O, -NR-C(=O)R-, -C(=O)NR-, -NR-C(=O)C(=O)NR-, -NR-C(=O)NR-, -NR-C(=S)NR-, -NR-C(=O)NRCH ₂ -, -NR-C(=S)NRCH ₂ -, wherein the R is selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted with 1-3 selected from the group N, S and A 3-12 membered heterocyclic group with a heteroatom of O;

M ¹ , M ² , M ³ or M ⁴ are each independently CR ¹ or N; and when said M ¹ , M ² , M ³ or M ⁴ is CR ¹ , said

It may be located on the CR ¹ (that is, the corresponding M ¹ , M ² , M ³ or M ⁴ is C at this time);

is a single or double bond;

^M is selected from the group consisting of CRa or NRa;

^M6 is selected from the group consisting of CR or NR;

^R is selected from the group consisting of H, CN, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, phenyl, substituted or unsubstituted C3-C8 cycloalkyl ;

Ring A is selected from the following group:

Wherein, V ¹ , V ² , V ³ , V ⁴ , V ⁵ , V ⁶ , V ⁷ or V ⁸ are each independently CR ² or N;

Each R ² is independently selected from the group consisting of H, CN, halogen, NH ₂ , substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, phenyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group with 1-3 heteroatoms selected from the group N, S and O, -C(O)-substituted or unsubstituted 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O, -(CH ₂ ) _m N(R ³ ) ₂ , -C(O)(CH ₂ ) _m N (R ³ ) ₂ , -NHC(O)(CH ₂ ) _m N(R ³ ) ₂ ; wherein, each of said R ³ is independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl , or two R ³ together with the connected N atoms form a 4-7 membered nitrogen-containing heterocyclic ring;

m is 0, 1, 2, 3, 4 or 5;

Ra has the structure shown in the following formula:

Wherein, the ring B is a substituted or unsubstituted C3-C8 cycloalkyl group, a substituted or unsubstituted 6-10 membered aryl group, a substituted or unsubstituted 5-12 membered heteroaryl group, a substituted or unsubstituted A 3-15 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O;

n is 0, 1, 2, 3, 4 or 5;

Each R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² are each independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy , substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 6-10 membered aryl, substituted or unsubstituted 3- with 1-3 heteroatoms selected from the group N, S and O 12-membered heterocyclic group; or said R ⁹ and R ¹⁰ or R ¹¹ and R ¹² together form a substituted or unsubstituted nitrogen atom with 1-3 heteroatoms selected from N, S and O -12 membered heterocyclyl;

Unless otherwise specified, the "substitution" refers to being substituted by one or more (such as 2, 3, 4, etc.) substituents selected from the following group: D, halogen, hydroxyl, C1-C6 alkoxy Group, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, methylsulfone, oxo (=O), -CN, -NH ₂ , -NHS (O) ₂ CH ₃ , C1-C6 amino group, carboxyl group, C1-C6 amido group (-C(=O)-N(Rd) ₂ or -NH-C(=O)(Rd), Rd is H or C1-C5 alkyl), or unsubstituted or substituted by one or more substituents selected from Z group, selected from the following groups: C1-C6 alkyl, C6-C10 aryl, with 1-3 5-10 membered heteroaryl, -(CH ₂ )-C6-C10 aryl, -(CH ₂ )-(having 1-3 heteroatoms selected from N, S and O 3-10 membered heteroaryl group with heteroatoms), -(5-10 membered heteroarylene group with 1-3 heteroatoms selected from N, S and O)-(C1-C6 alkyl), with 1-3 3-12-membered heterocyclic groups (including monocyclic, spiro, bridged or parallel rings) with 1-3 heteroatoms selected from the following group of N, S and O, and the group Z substituents are selected from the following Groups: halogen, C1-C6 alkyl, C1-C6 alkylene-OH, C1-C6 alkoxy, oxo, -S(O) ₂ CH ₃ , -CN, -OH, C6-C10 aryl, A 3-10 membered heterocyclic group having 1-3 heteroatoms selected from N, S and O, -C(O)CHNH ₂ , -C(O)CHOH;

Preferably, said X, Y, A, B, M ¹ , M ² , M ³ , m, n, p, q, Ra, Re, Rf, R ¹ , R ² , R ³ , R ⁴ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , and R ¹² are each independently the corresponding group in the specific compound in each embodiment.

The compounds of the present invention can act as HPK1 kinase inhibitors, in preferred embodiments, are HPK1 kinase selective inhibitors.

The preparation of formula I compound

Compounds of formula I of the present invention can be prepared by the following exemplary methods:

method 1:

R is various protecting groups attached to nitrogen, and the remaining protecting groups are as defined above.

Pharmaceutical compositions and methods of administration

Since the compound of the present invention has excellent HPK1 kinase inhibitory activity, the compound of the present invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and compounds containing the compound of the present invention as the main active ingredient The pharmaceutical composition of can be used to prevent and/or treat diseases related to HPK1 kinase activity or expression level (for example, cancer).

The pharmaceutical composition of the present invention comprises the compound of the present invention and pharmaceutically acceptable excipients or carriers in a safe and effective amount range. Wherein, "safe and effective dose" refers to: the amount of the compound is sufficient to obviously improve the condition without causing severe side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 10-200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use, and must have sufficient purity and low toxicity. "Compatibility" herein means that the components of the composition can be blended with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Examples of pharmaceutically acceptable carrier parts include cellulose and derivatives thereof (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid , magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween

), wetting agent (such as sodium lauryl sulfate), coloring agent, flavoring agent, stabilizer, antioxidant, preservative, pyrogen-free water, etc.

The administration method of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative administration methods include (but not limited to): oral administration, parenteral (intravenous, intramuscular or subcutaneous) injection and the like.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with (a) fillers or extenders, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, For example, glycerol; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow agents, such as paraffin; (f) Absorption accelerators such as quaternary ammonium compounds; (g) wetting agents such as cetyl alcohol and glyceryl monostearate; (h) adsorbents such as kaolin; and (i) lubricants such as talc, hard Calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shell materials, such as enteric coatings and others well known in the art. They may contain opacifying agents and, in such compositions, the release of the active compound or compounds may be in a certain part of the alimentary canal in a delayed manner. Examples of usable embedding components are polymeric substances and waxy substances. The active compounds can also be in microencapsulated form, if desired, with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, etc.

Besides such inert diluents, the compositions can also contain adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When administered in combination, the pharmaceutical composition also includes one or more other pharmaceutically acceptable compounds. One or more of the other pharmaceutically acceptable compounds may be administered simultaneously, separately or sequentially with the compound of the present invention.

When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage when administered, for a person with a body weight of 60kg, the daily The dosage is usually 1-2000 mg, preferably 20-500 mg. Of course, factors such as the route of administration and the health status of the patient should also be considered for the specific dosage, which are within the skill of skilled physicians.

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. For the experimental methods without specific conditions indicated in the following examples, the conventional conditions or the conditions suggested by the manufacturer are usually followed. Percentages are by weight and parts by volume unless otherwise indicated. Unless otherwise specified, the "normal phase column chromatography" purification mentioned herein refers to purification using normal phase silica gel column chromatography. The experimental materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified, or can be synthesized by adopting or following methods known in the art.

Synthetic example

Intermediate A1: 5-Chloro-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole-6-carbonitrile

tert-Butyl 6-bromo-5-chloro-1H-indazole-1-carboxylate

At room temperature, A1-0 (10.0 g, 43.2 mmol), N-ethyl-N-isopropylpropan-2-amine (8.54 g , 66.1 mmol) and dioxane (100 ml), and the stirrer was started. A solution of di-tert-butyldicarbonate (11.3 g, 51.8 mmol) in dioxane (20 mL) was then added dropwise to the reaction solution. After the dropwise addition, the reaction solution was stirred and reacted at room temperature for 12 hours. LCMS detected that the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-3:1) to obtain white solid A1-1 (12.56 g, yield: 87.7%). MS(ESI):m/z=274.9,276.9[M+H-56] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ8.58(s,1H),8.09(s,1H),7.83(s,1H),1.72(s,9H).

tert-Butyl 5-chloro-6-cyano-1H-indazole-1-carboxylate

Under argon protection, to a mixture of compound A1-1 (10.5 g, 31.67 mmol) and zinc cyanide (4.46 g, 38.0 mmol) in N,N-dimethylformamide (100 mL) was added tris( Dibenzylideneacetone) dipalladium (1.45 g, 1.58 mmol) and 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (1.82 g, 3.16 mmol). After replacing with argon three times, the reaction solution was heated to 100°C and stirred for 10 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with ethyl acetate (100 mL), and filtered through diatomaceous earth. The filtrate was then poured into water (400 mL), extracted with ethyl acetate (100 mL×3). The combined organic layers were washed successively with water (100 mL×5) and saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-4:1) to obtain white solid A1-2 (3.20 g, yield: 36.4%). MS(ESI):m/z=230.9,232.9[M+H-56] ⁺ .

¹ H NMR (400MHz, CDCl ₃ ) δ8.62(s, 1H), 8.20(d, J=0.8Hz, 1H), 7.89(d, J=0.4Hz, 1H), 1.74(s, 9H).

5-Chloro-1H-indazole-6-carbonitrile

A solution of A1-2 (3.20 g, 11.52 mmol) in 1,1,1,3,3,3-hexafluoropropan-2-ol (30 mL) was added to a 150 mL sealed tube. The reaction solution was heated to 110° C. and stirred for 2 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction was cooled to room temperature. Concentration under reduced pressure gave a crude product, which was purified by beating (petroleum ether/ethyl acetate=100:1-5:1, 20 mL) to obtain a white solid A1-3 (2.00 g, yield: 97.7%). MS(ESI):m/z=178.0,180.0[M+H] ⁺ .

5-Chloro-3-iodo-1H-indazole-6-carbonitrile

To a solution of A1-3 (2.0 g, 11.26 mmol) N,N-dimethylformamide (40 mL) was added N-iodosuccinimide (2.79 g, 12.39 mmol) at zero . After the reaction solution was stirred at room temperature for 2 hours, the reaction solution was placed in an 80°C oil bath, and the stirring was continued for 1 hour. After the reaction was cooled to room temperature, it was filtered, and the filter cake was washed three times with dichloromethane (10 mL). The resulting solid was dried in vacuo for 30 minutes to obtain light yellow solid A1-4 (3.4 g, yield: 99.4%). MS(ESI):m/z=304.0,306.0[M+H] ⁺ .

5-Chloro-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole-6-carbonitrile

Sodium hydride (purity: 60%, 538 mg, 13.44 mg Moore). After the reaction solution was stirred at zero for 30 minutes, (2-(chloromethoxy)ethyl)trimethylsilane (2.43 g, 14.56 mmol) was added dropwise thereto. After the dropwise addition, the ice bath was removed, and the reaction solution was stirred at room temperature for 2 hours. The reaction was detected by TLC. After the reaction was complete, the reaction solution was re-cooled to zero, and then a saturated ammonium chloride solution (100 ml) was slowly added to quench the reaction. The aqueous phase was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed successively with water (30 mL×3) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-4:1) to obtain light yellow solid A1 (3.3 g, yield: 67.9%). MS(ESI): m/z=434.0[M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ ) δ8.04-7.94 (m, 1H), 7.66 (d, J=0.4Hz, 1H), 5.73 (s, 2H), 3.52-5.58 (m, 2H), 0.94- 0.80(m,2H),-0.04(s,9H).

Intermediate A2: 5-Bromo-6-fluoro-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole

5-Bromo-6-fluoro-3-iodo-1H-indazole

To a solution of A2-0 (575 mg, 2.67 mmol) in acetonitrile (15 mL) was added N-iodosuccinimide (611 mg, 12.39 mmol) at room temperature. After the reaction mixture was stirred at room temperature for 2 hours, the reaction solution was placed in an 80°C oil bath, and stirred for 1 hour. After the reaction was cooled to room temperature, A2-1 (575 mg, yield: 57.8%) was obtained by filtration. MS(ESI): m/z=340.8,342.8[M+H] ⁺ .

5-Bromo-6-fluoro-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole

To a solution of A2-1 (545 mg, 1.60 mmol) in dry N,N-dimethylformamide (8.0 mL) was added sodium hydride (60%, 86 mg, 2.40 mmol) in portions at zero . After the reaction solution was stirred at zero for 20 minutes, a solution of (2-(chloromethoxy)ethyl)trimethylsilane (346 mg, 2.08 mmol) in N,N-dimethylformamide was added dropwise (0.5 ml). After the dropwise addition, the ice bath was removed, and the reaction solution was stirred at room temperature for 2 hours. The reaction was detected by TLC. After the reaction was complete, the reaction solution was cooled with an ice bath, and then a saturated ammonium chloride solution (10 mL) was slowly added dropwise to quench the reaction. The aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed successively with water (10 mL×3) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-6:1) to obtain light yellow solid A2-2 (503 mg, yield: 62.5%). MS (ESI): m/z = 471.4.4, 473.2 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ ) δ7.72(d, J=6.4Hz, 1H), 7.33(d, J=8.4Hz, 1H), 5.65(s, 2H), 3.58-3.49(m, 2H) ,0.91-0.83(m,2H),-0.05(s,9H).

The following compounds were obtained using a method similar to that of Example A1, replacing the corresponding raw materials.

Intermediate B1: (6-Methyl-5,6,7,8-tetrahydro-1,6-phthalazin-3-yl)boronic acid

3-Bromo-6-methyl-5,6,7,8-tetrahydro-1,6-naphthyridine

To B1-0 (0.50 g, 1.75 mmol) and paraformaldehyde (273 mg, 7.0 mmol) in methanol (10 mL) was added glacial acetic acid (210 mg, 3.5 mmol) at room temperature and stirred for 30 min Then, sodium cyanoborohydride (272 mg, 5.24 mmol) was added to the reaction solution. After the reaction solution was stirred at room temperature for 1 hour under the protection of nitrogen, the reaction solution was heated in a 65 degree oil bath and continued to stir for 1 hour. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a residue. Water (20 mL) was added to the residue, followed by extraction with dichloromethane (10 mL x 3). The combined organic phases were washed with saturated brine (10 mL), and the layers were separated. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse column chromatography to obtain yellow oil B1-1 (310 mg, yield: 78.1%). MS(ESI):m/z＝227.1,229.1[M+H] ⁺

(6-Methyl-5,6,7,8-tetrahydro-1,6-diazin-3-yl)boronic acid

Into a 50 ml round bottom flask, add B1-1 (310 mg, 1.37 mmol), double pinacol borate (693 mg, 2.73 mmol), [1,1-bis(diphenylphosphino ) ferrocene] palladium (II) dichloride (56 mg, 0.068 mmol), anhydrous potassium acetate (268 mg, 2.73 mmol) and anhydrous dioxane (5.0 mL). After replacing the reaction mixture with nitrogen three times, it was heated to 90° C. and stirred for 16 hours. The reaction was checked by LCMS. After the reactant was cooled to room temperature, dichloromethane (20 ml) was added to the reaction liquid, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. After the residue was dissolved in methanol, it was separated by reverse column chromatography to obtain pale yellow solid B1 (160 mg, yield: 61%). MS(ESI): m/z=193.5[M+H] ⁺ .

Intermediate B2: 2,5-Dimethyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3, 4-Tetrahydroisoquinoline

2-(4-Bromo-2-methylphenyl)ethane-1-amine hydrochloride

To a 250 mL round bottom flask was added B2-0 (5.00 g, 23.8 mmol), methanol (70 mL), di-tert-butyl dicarbonate (10.4 g, 47.6 mmol) and nickel chloride monohydrate in sequence (566 mg, 2.38 mmol). After the reaction mixture was reduced to zero degree, sodium borohydride (5.43 g, 142.8 mmol) was added slowly, and the reaction was stirred at 20 degree under nitrogen protection for 2 hours. LCMS detection reaction. After the conversion of the raw materials was complete, the reaction solution was poured into (200 mL) and extracted with dichloromethane (60 mL×3). The combined organic phases were washed with saturated brine (150 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a solid. This solid was dissolved in hydrogen chloride-dioxane solution (4N, 430 ml), and stirred at 20°C for 2 hours. The mixture was concentrated under reduced pressure to obtain B2-1 (5.18 g, yield 70.1%). MS(ESI):m/z=214.0,216.0[M+H] ⁺ .

N-(4-Bromo-2-methylphenethyl)formamide

A mixture of acetic anhydride (8.8 ml, 93.2 mmol) and formic acid (3.52 ml, 93.2 mmol) was stirred at 20° C. for 2 hours under nitrogen protection to obtain a mixed anhydride solution. Under nitrogen protection, a solution of B2-1 (4.68 g, 18.6 mmol) and triethylamine (5.2 mL, 37.2 mmol) in anhydrous tetrahydrofuran (50 mL) was cooled to zero, and then slowly added dropwise to the reaction solution A mixed anhydride solution. After the dropwise addition, the reaction solution was placed at room temperature and stirred for 16 hours under the protection of nitrogen. LCMS detected that the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain a residue. Water (80 mL) was added to the residue, followed by extraction with dichloromethane (30 mL×3). The combined organic phases were washed with saturated brine (30 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product B2-2 as brown oil (4.78 g, yield 100%). MS(ESI):m/z=242.0,244.0[M+H] ⁺ .

7-Bromo-5-methyl-3,4-dihydroisoquinoline-2(1H)-carbaldehyde

Under nitrogen protection, a solution of B2-2 (4.78 g, 18.6 mmol) in trifluoroacetic acid (30 mL) was added to a 100 mL round bottom flask equipped with a magnetic stirrer. Then paraformaldehyde (837 mg, 27.9 mmol) was added, and the reaction solution was heated to 75° C. and stirred for 3 hours. LCMS detection reaction. After the reaction was cooled to room temperature, the reaction solution was poured into water (100 mL), and extracted with dichloromethane (40 mL×3). The combined organic phases were washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=100:1-1:4) to obtain brown solid B2-3 (4.0 g, yield: 85%). MS(ESI):m/z=254.0,256.0[M+H] ⁺ .

7-bromo-5-methyl-1,2,3,4-tetrahydroisoquinoline

Dissolve B2-3 (2.0 g, 7.91 mmol) in a mixed solvent of methanol-water (20 mL/4.0 mL), then add sodium hydroxide (949 mg, 23.7 mmol), and stir the mixture at room temperature for 18 Hours, the reaction was detected by LCMS. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain a residue. Water (40 mL) was added thereto, followed by extraction with dichloromethane (15 mL×3). The combined organic phases were washed with saturated brine (40 mL), and the layers were separated. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a white solid crude product B2-4 (1.82 g, yield: 50.9%). MS(ESI):m/z=225.9,227.9[M+H] ⁺ .

7-bromo-2,5-dimethyl-1,2,3,4-tetrahydroisoquinoline

B2-4 (1.82 g, 8.05 mmol) was dissolved in methanol (15 mL), and paraformaldehyde (242 mg, 8.05 mmol) and sodium cyanoborohydride (556 mg, 8.85 mol), the reaction was stirred at room temperature for 2 hours under nitrogen protection. The reaction was checked by LCMS. After the reaction was completed, the solvent was concentrated under reduced pressure to obtain a residue. Water (30 mL) was added to the residue, followed by extraction with dichloromethane (15 mL x 3). The combined organic phases were washed with saturated brine (30 ml), and the layers were separated. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=100:1-1:2) to obtain yellow oil B2-5 (643 mg, yield: 33.2%). MS(ESI):m/z=240.0,242.0[M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ7.14(s,1H),7.02(s,1H),3.51(s,2H),2.69(m,4H),2.44(s,3H),2.18(s, 3H).

2,5-Dimethyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydro Isoquinoline

Into a 50 mL round bottom flask were added B2-5 (643 mg, 2.68 mmol), bis-linked pinacol borate (1.36 g, 5.35 mmol), [1,1-bis(diphenylphosphino) Ferrocene]palladium(II) dichloride (96 mg, 0.268 mmol), anhydrous potassium acetate (526 mg, 5.35 mmol) and anhydrous dioxane (5.0 mL). After the reaction mixture was replaced with nitrogen three times, it was heated to 90° C. and stirred for 5 hours. The reaction was checked by LCMS. After the reactant was cooled to room temperature, dichloromethane (20 ml) was added to the reaction liquid, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was dissolved in methanol and separated by reverse column chromatography to obtain light brown solid B2 (384 mg, yield: 50.0%). MS(ESI):m/z＝288.5[M+H] ⁺ .

Intermediate B3: 1-(2,6-Dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)- 4-Methylpiperazine

1-(4-Bromo-2,6-dimethylphenyl)-4-methylpiperazine

To B3-0 (4.88 g, 24.4 mmol) in diethylene glycol monomethyl ether (20 mL) was added 2-chloro-N-(2-chloroethyl)-N-methylethane at room temperature -1-Amine (4.7 g, 24.4 mmol). The reaction was heated to 160 degrees and stirred for 7 hours. After the reaction was detected by LCMS, after the reaction mixture was cooled to room temperature, the reaction solution was poured into aqueous sodium bicarbonate solution (250 mL), and extracted with dichloromethane (150 mL×2). The combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to remove the solvent to obtain an oily residue. The residue was separated by column chromatography (dichloromethane/methanol=100:1-7:3) to obtain B3-1 (2.2 g, yield: 32%). MS(ESI):m/z＝283.1,285.1[M+H] ⁺ .

1-(2,6-Dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-methyl Piperazine

To the mixed solvent of compound B3-1 (2.13 g, 7.55 mmol) in dioxane (20 ml) and water (4.0 ml), then add cesium carbonate (4.92 g, 15.1 mmol), bis-pina Alcohol borate (2.88 g, 11.3 mmol). After the mixture was replaced with nitrogen three times, it was placed in an oil bath at 100° C. and heated with stirring for 3 hours. After the reaction was detected by LCMS, the reaction solution was concentrated under reduced pressure. Water (100 mL) was added to the residue, followed by extraction with ethyl acetate (80 mL×2). The combined organic layers were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to remove the solvent to obtain a crude product. The crude product was separated by forward column chromatography (dichloromethane/methanol=100:1-3:7) to obtain B3 (1.7 g, yield: 47.8%). MS(ESI): m/z=331.0[M+H] ⁺ .

Intermediate B4: 6-isopropyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydro-4H -pyrazolo [1,5-d][1,4]diazoheptin-7(8H)-one

Methyl 2-(3-bromo-5-methyl-1H-pyrazol-1-yl)acetate

Dissolve B4-0 (9.00 g, 56 mmol), methyl chloroacetate (7.9 g, 72.8 mmol) and potassium carbonate (15 g, 112 mmol) in N,N-dimethylformamide (100 mL ), and the reaction was stirred at 30°C for 5 hours. LCMS detected that the reaction of the raw materials was complete, and the reaction solution was poured into water (500 mL), and extracted with ethyl acetate (500 mL×2). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=100:1-2:1) to obtain B4-1 (12.8 g, yield: 98%). MS(ESI):m/z=233.0,235.0[M+H] ⁺ .

Methyl 2-(3-bromo-5-(bromomethyl)-1H-pyrazol-1-yl)acetate

To B4-1 (6.8 g, 29 mmol) in carbon tetrachloride (60 mL) was added AIBN (480 mg, 2.9 mmol) at room temperature, and the solution was heated to 80°C after nitrogen replacement three times. Then NBS (5.7 g, 32 mmol) was added, and the reaction was stirred at 80° C. for 2 hours. LCMS detected that the reaction of raw materials was complete. After the reaction solution was cooled to room temperature, saturated sodium bicarbonate (200 ml) was added thereto for washing, the layers were separated, and the organic phase was collected. The aqueous phase was extracted with ethyl acetate (200 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude solid. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=100:1-5:1) to obtain B4-2 (5.4 g, yield: 59%). MS(ESI):m/z=312.8[M+H] ⁺ .

Methyl 2-(3-bromo-5-(cyanomethyl)-1H-pyrazol-1-yl)acetate

To B4-2 (5.3 g, 17 mmol) in dimethylsulfoxide (30 mL) was added sodium cyanide (1.25 g, 25.6 mmol) at room temperature. The reaction was stirred at 30°C for 1 hour. The reaction of the raw material was detected by LCMS to be complete. The reaction solution was poured into water (200 mL), extracted with ethyl acetate (200 mL×2). The combined organic phases were successively washed with water (100 mL×3), saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a crude solid. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=100:1-2:1) to obtain B4-3 (2.4 g, yield 55%). MS(ESI): m/z=257.9,259.9[M+H] ⁺ .

Methyl 2-(5-(2-aminoethyl)-3-bromo-1H-pyrazol-1-yl)acetate

To a solution of B4-3 (1.9 g, 7.4 mmol) in methanol (50 mL) was added platinum dioxide (168 mg, 0.74 mmol) at room temperature under argon protection. After cooling the reaction liquid to zero degree, trifluoroacetic acid (2.9 g, 25.9 mmol) was added. After three displacements of hydrogen, the reaction was stirred at room temperature under an atmosphere of hydrogen (15 psi) for 16 hours. The reaction was detected by LCMS, and the raw material was completely reacted. After filtration, the filtrate was concentrated under reduced pressure to obtain B4-4 (2 g, crude product). MS(ESI):m/z=262.0,264.0[M+H] ⁺ .

2-Bromo-5,6-dihydro-4H-pyrazolo[1,5-d][1,4]diazoheptin-7(8H)-one

To a solution of B4-4 (2.00 g) in methanol (20 mL) was added triethylamine (5.0 mL) under argon protection, and the reaction was stirred at 30°C for 1 hour. As detected by LCMS, the raw materials were completely reacted. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was separated and purified by silica gel column (dichloromethane/methanol=100:1-40:1) to obtain compound B4-5 (820 mg, yield: 46%). MS(ESI):m/z=230.0,232.0[M+H] ⁺ .

2-Bromo-6-isopropyl-5,6-dihydro-4H-pyrazolo[1,5-d][1,4]diazoheptin-7(8H)-one

To a solution of B4-5 (475 mg, 2.1 mmol) in N,N-dimethylformamide (10 mL) was added portionwise sodium hydride (198 mg, 8.4 mmol) under cooling in an ice-water bath. After the reaction solution was stirred at zero for 30 minutes, 2-iodopropane (3.57 g, 21.0 mmol) was added dropwise thereto. After the dropwise addition, the reaction solution was slowly raised to room temperature, and then continued to stir for 2 hours. Complete conversion of starting material was detected by LCMS. Water (20 ml) was added to the reaction solution, solids were precipitated, filtered, and the filter cake was purified by silica gel column (dichloromethane/methanol=100:1-20:1) to obtain B4-6 (220 mg, yield: 39 %). MS(ESI):m/z=272.0,274.0[M+1] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ6.02(s,1H),4.99(s,2H),4.76(dt,J=13.6,6.8Hz,1H),3.65-3.55(m,2H),3.03- 2.94(m,2H),1.11(d,J＝6.8Hz,6H).

6-isopropyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydro-4H-pyrazolo [1,5-d][1,4]diazo Heptin-7(8H)-one

Under argon protection, B4-6 (250 mg, 0.918 mmol), bis-boronic acid pinacol ester (700 mg, 2.76 mmol), [1,1-bis( Diphenylphosphino)ferrocene]palladium(II) dichloride (75 mg, 0.092 mmol), anhydrous potassium acetate (180 mg, 1.84 mmol) and anhydrous dioxane (5.0 mL). After the reaction mixture was replaced with argon three times, it was heated to 100°C and stirred for 16 hours. The reaction was checked by LCMS. After the reactant was cooled to room temperature, it was filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was separated by reverse column chromatography to obtain B4 (101 mg, yield: 46.4%). MS(ESI):m/z＝238.1[M+H] ⁺ .

Intermediate B5: Methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroisoquinoline-1 (2H)-Kone

7-Bromo-2-methyl-3,4-dihydroisoquinolin-1(2H)-one

To a solution of B5-0 (300 mg, 1.33 mmol) in N,N-dimethylformamide (10 mL) was added sodium hydride (60%, 64 mg, 1.60 mmol) at zero degrees. After the reaction solution was stirred at this temperature for 30 minutes, iodomethane (226.5 mg, 1.60 mmol) was added dropwise, and then the reaction was warmed to room temperature and stirred for 2 hours. LCMS detection reaction. After the reaction was completed, water (30 mL) was added to quench the reaction, and then extracted with ethyl acetate (10 mL×3). The combined organic phases were washed successively with water (10 mL×3), saturated brine (10 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography to obtain B5-1 (318 mg, 1.32 mmol). MS(ESI):m/z=240.0,242.0[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ8.04 (d, J = 2.0Hz, 1H), 7.61 (dd, J = 8.0, 2.0Hz, 1H), 7.21 (d, J = 8.0Hz, 1H), 3.62(t,J=6.8Hz,2H),3.14(s,3H),3.00(t,J=6.8Hz,2H).

Methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroisoquinoline-1(2H)- ketone

Under the protection of argon, to B5-1 (318 mg, 1.33 mmol), bis-boronic acid pinacol ester (1.01 g, 3.99 mmol), [1,1'-bis (diphenylphosphine) diphenocene Iron]Palladium dichloride dichloromethane complex (109 mg, 0.13 mmol) and anhydrous potassium acetate (391 mg, 3.99 mmol) were added 1,4-dioxane (2.5 mL) . After argon replacement three times, the reaction was heated to 90°C and stirred for 16 hours. The reaction was checked by LCMS. After filtration, the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-5:1) to obtain B5 (300 mg, yield: 78.5%). MS(ESI):m/z＝288.2[M+H] ⁺ .

The following compounds were obtained using a method similar to that of Example B5, replacing the corresponding raw materials.

Intermediate B10: 2-methyl-1-(5-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-3, 4-Dihydroisoquinoline- 2(1H)-yl)propan-2-ol

1-(7-Bromo-5-methyl-3,4-dihydroisoquinolin-2(1H)-yl)-2-methylpropan-2-ol

To a solution of 7-bromo-5-methyl-1,2,3,4-tetrahydroisoquinoline (133 mg, 0.59 mmol) in ethanol (5 mL) was added 2,2-dimethyl Dioxaline (190.6 mg, 2.65 mmol). The reaction solution was heated to 80°C and stirred for 12 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain crude product B10-1 (176 mg). The crude product was directly used in the next step without isolation and purification. MS(ESI):m/z=298.0,300.0[M+H] ⁺ .

2-Methyl-1-(5-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4 dihydro Isoquinolin-2(1H)-yl)propane alkan-2-ol

Under argon protection, the crude product compound B10-1 (176 mg), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1, 3,2-dioxaborolane) (599 mg, 2.36 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (48.3 mg, 0.059 mmol) and potassium acetate (173.5 mg, 1.77 mmol) was added 1,4-dioxane (5 mL). After argon replacement three times, the reaction solution was heated to 90°C and stirred for 4 hours. The reaction was checked by LCMS. After the reaction was completed, ethyl acetate (10 mL) was added to dilute the reaction solution, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse column chromatography to obtain B10 (129 mg, yield: 63.3%). MS(ESI): m/z=346.2[M+H] ⁺ .

Intermediate C1: tert-butyl (3‐fluoro‐5‐methyl‐4‐(4,4,5,5‐tetramethyl‐1,3,2‐dioxolan‐2‐yl)benzyl) (Methyl)amino Formate

4-Bromo-3-fluoro-5-methylaniline

To a solution of C1-0 (5.00 g, 39.95 mmol) in N,N-dimethylformamide (40 mL) was added N-bromosuccinimide (7.25 g, 40.75 Moore). After the addition, the reaction solution was stirred at 0°C for 30 minutes, then the ice bath was removed, and the temperature was raised to room temperature to continue stirring for 1 hour. The reaction was checked by LCMS. After the reaction, water (120 mL) was added to the reaction solution, followed by extraction with ethyl acetate (50 mL×3). The combined organic phases were successively washed with saturated aqueous sodium bicarbonate (50 mL), water (50 mL×3), and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-3:1) to obtain C1-1 (7.9 g, yield: 96.9%). MS(ESI):m/z=204.0,206.0[M+H] ⁺ .

2-Bromo-1-fluoro-5-iodo-3-methylbenzene

To a solution of compound C1-1 (8.0 g, 40.0 mmol) in acetonitrile (200 mL) was added 20% aqueous sulfuric acid (99 mmol of concentrated sulfuric acid diluted in 20 mL of water) at zero temperature. After stirring for 5 minutes, a solution of sodium nitrite (5.4 g, 78.0 mmol, in 120 ml of water) was added dropwise to the reaction solution, and the addition was completed in 15 minutes. Under ice-cooling, potassium iodide (25.6 g, 158 mmol) was added, and then the reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, sodium sulfite was added to the reaction liquid to quench the reaction. Add ethyl acetate (100 ml) for extraction, and separate the layers. The organic phase was washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=100:1-1:10) to obtain C1-2 (11.2 g, yield: 89%).

¹ H NMR (400MHz, CDCl ₃ )δ7.39(m,1H),7.29(m,1H),2.38(s,3H).

2-Bromo-1-fluoro-3-methyl-5-vinylbenzene

At room temperature, to compound C1-2 (7.34 g, 47.6 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxolane (10 g, 31.7 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (2.32 g, 3.17 mmol) and potassium phosphate (13.4 g, 63.5 mmol), adding 1,4-Dioxane (100ml) and water (10ml). After nitrogen replacement three times, the reaction was heated to 70°C and stirred for 16 hours. As detected by TLC, a new point was formed in the reaction. After filtration, the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column separation (petroleum ether=100%) to obtain colorless oil C1-3 (5.6 g, yield: 81%).

4-Bromo-3-fluoro-5-methylbenzaldehyde

At zero degrees, C1-3 (5.40 g, 25.1 mmol) was dissolved in a mixed solvent of tetrahydrofuran (120 ml) and water (30 ml), and potassium dihydrate (0.925 g, 2.51 mmol) was added successively. Sodium periodate (21.4 g, 100 mmol). The reaction was stirred at 15°C for 2 hours. The reaction was detected by thin layer chromatography. After the reaction was completed, the insoluble matter was filtered off, and the filtrate was poured into water (100 mL), followed by extraction with dichloromethane (100 mL×2). The combined organic layers were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column (petroleum ether/ethyl acetate=100:1-10:1) to obtain C1-4 (4.70 g, yield: 96%).

1-(4-Bromo-3-fluoro-5-methylphenyl)-N-methylmethylamine

Compound C1-4 (4.70 g, 21.6 mmol) was dissolved in methanol (70 ml), and after stirring at 15°C for 30 minutes, methylamine hydrochloride (7.31 g, 108 mmol) and cyanoboron were added successively Sodium hydride (5.44 g 86.6 mmol). The reaction was stirred at room temperature for 16 hours. Thin-layer chromatography detected that the reaction was complete. Then, saturated sodium bicarbonate solution (100 ml) was added for washing, and dichloromethane was added for extraction (100 ml×3). The combined organic layers were washed with saturated brine (150 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain C1-5 (4.90 g, yield: 97%). MS(ESI):m/z=232.1,234.1[M+H] ⁺ .

tert-Butyl(4-bromo-3-fluoro-5-methylbenzyl)(methyl)carbamate

To a solution of C1-5 (4.9 g, 21.1 mmol) and triethylamine (6.41 g, 63.3 mmol) in dichloromethane (60 mL) was added di-tert-butyl dicarbonate (13.8 g, 63.3 mmol) at room temperature. mol), the reaction mixture was stirred at room temperature for 16 hours. The reaction was detected by thin layer chromatography. After conversion of the starting material, water (100 mL) was added to the reaction mixture for washing. The organic phase was separated, and the aqueous phase was extracted with dichloromethane (100 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-10:1) to obtain C1-6 (1.88 g, yield: 26%). MS(ESI):m/z=275.9,277.7[M+H-56] ⁺ .

tert-butyl(3-fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxolan-2-yl)benzyl)(methyl) Urethane

Under nitrogen protection, compound C1-6 (1.68 g, 5.04 mmol), biboronic acid pinacol ester (5.13 g, 20.2 mmol), [1,1'-bis(diphenylphosphino) diphenocene To a mixture of iron]palladium dichloride (411 mg, 0.504 mmol) and potassium acetate (3.21 g, 15.2 mmol), 1,4-dioxane (30 mL) was added. After the mixture was replaced with nitrogen three times, the reaction solution was placed in a 100°C oil bath and stirred for 16 hours. LCMS detection reaction. After the reaction was completed, it was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse phase column to obtain intermediate C1 (1.4 g, yield: 72%). MS(ESI):m/z=380.2[M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ6.68(m,2H),4.30(m,2H),2.85(s,3H).2.42(s,3H),1.45(s,9H),1.37(s, 12H)

Intermediate C2: Methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroisoquinoline-1 (2H)-Kone

3-N,N-di-tert-butoxycarbonyl 5-bromo-4-methylpyridine

To a solution of C2-0 (1.00 g, 5.35 mmol) in dichloromethane (40 mL), di-tert-butyl dicarbonate (2.45 g, 11.24 mmol) and 4-dimethylaminopyridine (32.7 mg, 0.27 mmol), the reaction system was stirred at room temperature for 16 hours. The reaction solution was distilled off the solvent under reduced pressure to obtain an oil, which was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-6:1) to obtain a colorless oil C2-1 (860 mg, Yield: 41.5%). MS(ESI):m/z=387.1,389.1[M+H] ⁺ .

4-N,N-bis-tert-butoxycarbonyl 4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine

Under argon protection, in a 50 ml round bottom flask equipped with a magnetic stirrer, add C2-1 (860 mg, 2.22 mmol), bisanalyl borate (2.82 g, 11.1 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (362.6 mg, 0.44 mmol), potassium acetate (653 mg, 6.66 mmol) and anhydrous 1 , 4-dioxane (10 ml). After argon replacement three times, the reaction was placed in an oil bath and stirred at 90°C for 3.5 hours. The reaction was detected by TLC. After the reaction was cooled to room temperature, ethyl acetate (20 mL) was added to dilute the reaction solution, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-5:1) to obtain yellow oil C2 (898 mg, yield: 93.2%). MS(ESI): m/z=435.2[M+H] ⁺ .

Intermediate C3: 6,8-difluoro-7-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-3,4-dihydroisoquine Phenyl-2(1H)-carboxylate tert-butyl acid

2-(3,5-Difluorophenyl)ethyl-1-amine hydrochloride

To a mixed solution of C3-0 (5.0 g, 32.65 mmol) in aqueous hydrochloric acid (4.0 mL) and methanol (60 mL) was added Pd/C (10%, 500 mg) at room temperature. The reaction mixture was stirred at room temperature for 5 hours under an atmosphere of hydrogen (45 psi). TLC showed the reaction was complete. The reaction solution was diluted with 50 ml of methanol, filtered through celite, and the filter cake was washed with 50 ml of methanol. The filtrate was concentrated in vacuo to obtain crude product C3-1 (6.3 g, yield: 100%). MS(ESI):m/z=158.1[M+H] ⁺ .

¹ H NMR (400MHz,DMSO-d ₆ )δ8.09(br s,3H),7.13(m,1H),7.09-7.01(m,2H),3.12-3.02(m,2H),2.96-2.89( m,2H).

N-(3,5-Difluorophenethyl)-2,2,2-trifluoroacetamide

To a mixture of C3-1 (6.30 g, 32.54 mmol) in ethyl acetate (60 mL) was added triethylamine (22.6 mL, 162.7 mmol) at room temperature, and then trifluoroacetic anhydride (6.0 mL, 42.3 mmol). The reaction mixture was stirred at room temperature for 5 hours. TLC showed the reaction was complete. The reaction solution was diluted with ethyl acetate (50 mL), and washed with saturated brine (50 mL×2). The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain C3-2 (8.2 g, yield: 100%).

1H NMR (400MHz, CDCl ₃ )δ6.94-6.78(m,1H),6.75-6.63(m,3H),3.59(q,J=6.8Hz,2H),2.88(t,J=7.2Hz,2H ).

1-(6,8-Difluoro-3,4-dihydroisoquinolin-2(1H)-yl)-2,2,2-trifluoroethyl-1-one

To a solution of C3-2 (4.00 g, 15.8 mmol) in glacial acetic acid (40 mL) was slowly added concentrated sulfuric acid (20 mL) at 5°C, followed by paraformaldehyde (1.42 g, 15.8 mmol). The reaction mixture was warmed slowly and stirred at room temperature for 5 hours. TLC showed the reaction was complete. The reaction mixture was poured into vigorously stirred ice water (200 mL), neutralized with sodium bicarbonate, and extracted with ethyl acetate (200 mL). The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-6:1) to obtain C3-3 (2.35 g, yield: 56%).

¹ H NMR (400MHz, CDCl ₃ )δ6.77-6.67(m,2H),4.78-4.69(m,2H),3.93-3.80(m,2H),2.99-2.89(m,2H).

6,8-Difluoro-1,2,3,4-tetrahydroisoquinoline

At room temperature, anhydrous potassium carbonate (3.67 g, 26.6 mmol) was added to a mixture of C3-3 (2.35 g, 8.86 mmol) in water (10 ml) and methanol (30 ml), and the reaction mixture was heated at 60° C. Stir for 5 hours. TLC showed detection reaction. After the reaction was complete, the solution was concentrated under reduced pressure to obtain a residue, which was then extracted with ethyl acetate-methanol mixed solution (50 mL, 10:1). The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain C3-4 (1.38 g, yield: 92%). MS(ESI):m/z=170.1[M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ ) δ6.66-6.55(m, 2H), 3.96(s, 2H), 3.09(t, J=6.0Hz, 2H), 2.76(t, J=5.6Hz, 2H) ,1.71(br s,1H).

tert-butyl 6,8-difluoro-3,4-dihydroisoquinoline-2(1H)-carboxylate

To a solution of C3-4 (1.38 g, 8.15 mmol) and triethylamine (1.7 mL, 12.2 mmol) in ethyl acetate (30 mL) was added di-tert-butyl dicarbonate (2.14 g, 9.79 mmol) at room temperature. Moore). The reaction mixture was stirred at room temperature for 5 hours. The reaction was detected by TLC. The reaction mixture was diluted with ethyl acetate (20 mL) and washed with water (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain C3-5 (2.11 g, yield: 96%). MS (ESI): m/z = 214.0 [M+H-56] ⁺ .

6,8-Difluoro-7-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-3,4-dihydroisoquinoline-2( 1H)-tert-butyl carboxylate

To a solution of C3-5 (2.50 g, 9.28 mmol) in anhydrous methyl tetrahydrofuran (40 mL) was slowly added dropwise lithium diisopropylamide (11.6 mL, 23.2 mmol) at -65°C. After the dropwise addition, the reaction mixture was stirred at -65° C. for 1 hour under the protection of nitrogen. Then 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborane (6.6 mL, 32.5 mmol) was slowly added dropwise to the reaction solution. The mixture was stirred at -65°C for 1 hour, then slowly warmed to room temperature and stirred for 2 hours. The reaction was checked by TLC. Water (40 mL) was added to quench the reaction, then extracted with ethyl acetate (80 mL). The organic phase was concentrated under reduced pressure to obtain a crude product, which was purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-6:1) to obtain C3 (2.05 g, yield: 55.9%). MS(ESI):m/z=340[M+H-56] ⁺ .

1H NMR (400MHz, CDCl ₃ ) δ6.65(d, J=9.2Hz, 1H), 4.51(s, 2H), 3.62(t, J=5.6Hz, 2H), 2.80(t, J=5.2Hz, 2H), 1.48(s,9H), 1.37(s,12H).

Intermediate C4: 2-(dimethylamino)-N-(3-fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane -2-yl)benzene base) acetamide

4-Bromo-3-fluoro-5-methylaniline

To a solution of C1-0 (5.00 g, 40.0 mmol) in N,N-dimethylformamide (40 mL) was added N-bromosuccinimide (7.25 g, 40.75 Moore). The reaction was stirred at zero degrees for 30 minutes, then the ice bath was removed and allowed to warm to room temperature and stirring was continued for 1 hour. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was poured into water (100 mL), and extracted with ethyl acetate (40 mL×3). The combined organic phases were successively washed with saturated aqueous sodium bicarbonate (50 mL), water (50 mL×3), brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-3:1) to obtain off-white solid C4-1 (7.9 g, yield: 96.9%). MS(ESI):m/z=203.0,205.0[M+H] ⁺ .

(4-Bromo-3-fluoro-5-methylphenyl)-2-chloroacetamide

To C4-1 (612 mg, 3.0 mmol), 2-chloroacetic acid (284 mg, 3.0 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'- In a solution of tetramethylurea hexafluorophosphate (1.71 g, 4.5 mmol) in dichloromethane (7.0 ml), N,N-diisopropylethylamine (774 mg, 6.0 mmol) was added, and the reaction solution Stir at room temperature for 2 hours. The reaction was checked by LCMS. After the reaction was completed, water (10 ml) was added to quench the reaction, the layers were separated, and the aqueous phase was extracted with dichloromethane (5 ml×2). The combined organic phases were washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-2:1) to obtain light yellow solid C4-2 (504 mg, yield: 60.0%). MS(ESI):m/z=280.0,282.0[M+H] ⁺ .

N-(4-bromo-3-fluoro-5-methylphenyl)-2-(dimethylamino)acetamide

To a solution of C4-2 (841 mg, 3.0 mmol) in N,N-dimethylformamide (8 mL) was added dimethylamine in THF (2.0 mol/L, 3.0 mL, 6.0 mmol) at room temperature Solution, the temperature of the reaction solution was raised to 80°C and stirred for 2 hours. The reaction was checked by LCMS. After the reaction was completed, it was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (dichloromethane/methanol=100:1-15:1) to obtain light yellow oil C4-3 (280 mg, yield: 32.3%). MS(ESI):m/z=289.0,291.0[M+H] ⁺ .

(Dimethylamino)-N-(3-fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl ) phenyl) acetamide

Under the protection of argon, to C4-3 (140 mg, 0.48 mmol) and), bisanalyl borate (614.5 mg, 2.42 mmol), [1,1'-bis (diphenylphosphine ) ferrocene] dichloropalladium dichloromethane complex (79.2 mg, 0.097 mmol) and anhydrous potassium acetate (142.3 mg, 1.45 mmol) mixture, add anhydrous dioxane (1.5 ml ). After argon replacement three times, the reaction solution was heated to 100°C and stirred for 12 hours. The reaction was checked by LCMS. After the reaction was completed, ethyl acetate (10 mL) was added to dilute the reaction solution, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (dichloromethane/methanol=100:1-15:1) to obtain yellow solid C4 (148 mg, yield: 73%). MS(ESI):m/z=337.2[M+H] ⁺ .

Intermediate C5: tert-butyl 2-(3-fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl ) phenyl) piperazine Pyridine-1-carboxylate

tert-butyl 6-(4-bromo-3-fluoro-5-methylphenyl)-3,4-dihydropyridine-1(2H)-carboxylate

At room temperature, to C1-2 (500 mg, 1.59 mmol), N-tert-butoxycarbonyl-3,4-dihydropyridine-6-boronic acid pinacol ester (490 mg, 1.59 mmol), [1 ,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (116 mg, 0.159 mmol) and anhydrous potassium carbonate (438 mg, 3.18 mmol), add 1,4-di Hexane (10ml) and water (4ml). After the reaction mixture was replaced with nitrogen three times, under the protection of nitrogen, it was heated to 65° C. and stirred for 16 hours. TLC detection reaction. The reaction mixture was concentrated, added with water (5 mL), and extracted with ethyl acetate (20 mL×2). The combined organic phases were concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-6:1) to obtain C5-1 (265 mg, yield: 45%). MS(ESI):m/z=313.9,315.9[M+H-56] ⁺ .

6-(4-Bromo-3-fluoro-5-methylphenyl)-1,2,3,4-tetrahydropyridine

To a solution of C5-1 (265 mg, 0.715 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (0.53 mL, 2.85 mmol) at room temperature. After the reaction solution was stirred at room temperature for 2 hours, the reaction was concentrated under reduced pressure to obtain C5-2 (193 mg, yield: 100%). MS(ESI):m/z=270.0,272.0[M+H] ⁺ .

2-(4-Bromo-3-fluoro-5-methylphenyl)piperidine

To a solution of C5-2 (193 mg, 0.71 mmol) and triethylamine (287 mg, 2.84 mmol) in THF (5 mL) was added sodium cyanoborohydride (90 mg, 1.43 mmol) at room temperature . The reaction body was stirred at room temperature for 3 hours. After the reaction solution was concentrated, ethyl acetate (20 mL) and water (10 mL) were added to stir, extract, and collect the organic phase. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain C5-3 (194 mg, yield: 100%). MS(ESI):m/z=274.0,274.0[M+H] ⁺ .

tert-butyl 2-(4-bromo-3-fluoro-5-methylphenyl)piperidine-1-carboxylate

To a solution of C5-3 (190 mg, 0.698 mmol) and triethylamine (106 mg, 10.5 mmol) in ethyl acetate (10 mL) was added di-tert-butyl dicarbonate (183 mg, 0.838 Millimoles). After the reaction mass was stirred at room temperature for 2 hours, TLC showed that the reaction was complete. The reaction solution was diluted with ethyl acetate (10 mL), washed with water (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-6:1) to obtain C5-4 (260 mg, yield: 100%). MS(ESI):m/z=315.9,317.9[M+H-56] ⁺ .

¹ H NMR (400MHz, CDCl ₃ ) δ6.86(s, 1H), 6.81(d, J=9.6Hz, 1H), 5.34-5.30(m, 1H), 4.04(d, J=12.8Hz, 1H) ,2.78-2.67(m,1H),2.41(s,3H),2.17-2.24(m,1H),1.93-1.81(m,1H),1.61-1.53(m,2H),1.46(s,9H) ,1.42-1.23(m,2H).

tert-butyl 2-(3-fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl) piperidine-1-carboxylate

Under the protection of argon, C5-4 (200 mg, 0.54 mmol), bisanalyl borate (546 mg, 2.15 mmol), [1,1'-bis(diphenylphosphine) di Ferrocene] dichloropalladium dichloromethane complex (43 mg, 0.053 mmol) and anhydrous potassium acetate (211 mg, 2.15 mmol) mixture, add anhydrous 1,4-dioxane ( 3.0 ml). After argon replacement three times, the reaction solution was heated to 100°C and stirred for 16 hours. The reaction was checked by LCMS. After the reaction was completed, ethyl acetate (10 mL) was added to dilute the reaction solution, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse column chromatography to obtain C5 (132 mg, yield: 58.6%). MS(ESI):m/z=364.1[M+H-56] ⁺ .

Example 1: 1-(4-(3-(2,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-6-isocyano-1H-indazole -5-yl)-3-fluoro- 5-methylphenyl)-N-methylmethylamine

5-Chloro-3-(2,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-1-((2-(trimethylsilyl)ethoxy Base) methyl) -1H- Indazole-6-carbonitrile

Under argon protection, add A1 (250 mg, 0.576 mmol), B2 (199 mg, 0.691 mmol), potassium phosphate (245 mg, 1.15 mmol) and 1,4-dioxane (4.5 mL) To a mixture of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (42 mg, 0.057 mmol) was added to a mixture of water (1.5 mL). After replacing with argon three times, the reaction solution was heated to 80°C and stirred for 3 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (15 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-1:99) to obtain light brown solid compound 1-1 (260 mg, yield: 96%). MS(ESI): m/z=467.2,469.2[M+H] ⁺ .

tert-Butyl(4-(3-(2,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-6-isocyano-1-(2-(tri Methylsilyl)ethoxy Base) methyl) -1H-indazol-5-yl) -3-fluoro-5-methylbenzyl) (methyl) carbamate

Under argon protection, to compound 1-1 (100 mg, 0.214 mmol), tert-butyl (3-fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1, 3,2-dioxolan-2-yl)benzyl)(methyl)carbamate (97 mg, 0.257 mmol), potassium phosphate (91 mg, 0.428 mmol) and 1,4-dioxo To a mixture of hexacyclic (1.5 ml)/water (0.5 ml), methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-bis Phenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (17 mg, 0.02 mmol). Under the protection of argon, the reaction solution was heated to 100°C and stirred for 3 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (5 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=4:1-1:99) to obtain light yellow solid compound 1-2 (83 mg, yield: 56.7%). MS(ESI):m/z＝685.0[M+H] ⁺ .

1-(4-(3-(2,5-Dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-6-isocyano-1H-indazol-5-yl )-3-fluoro-5-methylphenyl)- N-Methylmethylamine

Add compound 1-2 (83 mg, 0.121 mmol) and anhydrous dichloromethane (2.0 ml) successively to a 50 ml round bottom flask equipped with a magnetic stirrer, start the stirrer, and wait for the reaction solution to cool to zero. Trifluoroacetic acid (1.0 mL) was added to the reaction vial. The ice bath was removed, and the reaction was stirred at room temperature for 4 hours. The solution was concentrated under reduced pressure to obtain an oily residue, to which anhydrous methanol (3.0 mL) was added. After cooling to zero with an ice-water bath, anhydrous potassium carbonate was added to the reaction solution to adjust the pH to 10-11. Start the stirrer, stir for 30 minutes, filter, and concentrate the filtrate under reduced pressure to obtain the crude product. The crude product was separated and purified by reverse column chromatography to obtain the target product 1 (16 mg, yield: 29.1%) as a white solid. MS(ESI): m/z=454.1[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ8.21(s,1H),8.06(s,1H),7.76(s,1H),7.61(s,1H),7.35(s,1H),7.27(d ,J=9.2Hz,1H),4.59-4.46(m,2H),4.24(s,2H),3.14(t,J=6.4Hz,2H),3.05(s,3H),2.77(s,3H) ,2.37(s,3H),2.20(s,3H).

Example 2: 3-(5-(2-fluoro-6-methoxyphenyl)-6-isocyano-1H-indazol-3-yl)-6-methyl-5,6,7,8 -tetrahydro-1,6-di Azine

5-Chloro-3-(6-methyl-5,6,7,8-tetrahydro-1,6-naphthyridine-3-yl)-1-((2-(trimethylsilyl )ethoxy)methyl)- 1H-Indazole-6-carbonitrile

Under argon protection, compound A1 (200 mg, 0.461 mmol), B2 (106 mg, 0.553 mmol), potassium phosphate (196 mg, 0.992 mmol) and 1,4-dioxane (3.0 mL )/water (1.0 mL), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (37 mg, 0.046 mmol) was added. Under the protection of argon, the reaction solution was heated to 90°C and stirred for 3 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (10 mL×4). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-1:99) to obtain light brown solid compound 2-1 (135 mg, yield: 64.5%). MS(ESI): m/z=454.1,456.1[M+H] ⁺ .

3-(5-(2-fluoro-6-methoxyphenyl)-6-isocyano-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole -3-base)-6- Methyl-5,6,7,8-tetrahydro-1,6-naphthyridine

Under argon protection, compound 2-1 (65 mg, 0.143 mmol), (2-fluoro-6-methoxyphenyl) boronic acid (49 mg, 0.286 mmol), potassium phosphate (76 mg, 0.358 mmol) mol) and 1,4-dioxane (1.2 ml)/water (0.4 ml) mixture was added methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-iso Propyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (11.8 mg, 0.014 mmol). Under the protection of argon, the reaction solution was heated to 90°C and stirred for 3 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction liquid was cooled to room temperature, and ethyl acetate (5 mL×3) was added for extraction. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse silica gel column to obtain off-white solid compound 2-2 (59 mg, yield: 57.3%). MS(ESI): m/z=544.1[M+H] ⁺ .

3-(5-(2-fluoro-6-methoxyphenyl)-6-isocyano-1H-indazol-3-yl)-6-methyl-5,6,7,8-tetrahydro- 1,6-naphthyridine

To compound 2-2 (59 mg, 0.108 mmol) in anhydrous dichloromethane (2.0 mL) was added trifluoroacetic acid (1.0 mL) under ice-cooling. The ice bath was removed, and the reaction was stirred at room temperature for 4 hours. The solution was concentrated under reduced pressure to obtain an oily residue, which was then dissolved in anhydrous methanol (3.0 mL). After the solution was cooled to zero with an ice-water bath, anhydrous potassium carbonate was added to the reaction solution to adjust the pH to 10-11. Start the stirrer, stir for 30 minutes, filter, and concentrate the filtrate under reduced pressure to obtain the crude product. The crude product was separated by preparative high performance liquid chromatography to obtain white solid compound 2 (30 mg, yield: 66.9%). MS(ESI):m/z=414.1[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ8.89(d, J=2.0Hz, 1H), 8.12(s, 1H), 8.06(d, J=2.0Hz, 1H), 8.04(s, 1H), 7.45(d,J=6.8Hz,1H),6.97(d,J=8.8Hz,1H),6.87(s,1H),3.79(s,3H),3.73(s,2H),3.09(d,J =6.0Hz, 2H), 2.89(t, J=6.0Hz, 2H), 2.50(s, 3H).

Example 3: 5-(6,8-difluoro-1,2,3,4-tetrahydroisoquinolin-7-yl)-3-(2,5-dimethyl-1,2,3, 4-Tetrahydroisoquinoline-7- base)-1H-indazole-6-carbonitrile

tert-Butyl 7-(6-cyano-3-(2,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-1-((2-(tri Methylsilyl)ethoxy Base)methyl)-1H-indazol-5-yl)-6,8-difluoro-3,4-dihydroisoquinoline-2(1H)-carboxylate

Under argon protection, compound 1-1 (80 mg, 0.17 mmol), C3 (81 mg, 0.21 mmol), [1,1'-bis(diphenylphosphino)ferrocene]-di To a mixture of palladium(II) chloride dichloromethane complex (15 mg, 0.017 mmol) and potassium phosphate (109 mg, 0.51 mmol), dioxane (1.0 mL) and water (0.3 mL) were added . Under argon protection, the reaction was heated to 100°C and stirred for 3 hours. The reaction was checked by LCMS. After the reaction solution was cooled to room temperature, ethyl acetate (10 mL×2) was added for extraction. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (dichloromethane/methanol=100:1-20:1) to obtain brown solid compound 3-1 (120 mg). MS(ESI):m/z=700.3[M+H] ⁺ .

5-(6,8-difluoro-1,2,3,4-tetrahydroisoquinolin-7-yl)-3-(2,5-dimethyl-1,2,3,4-tetrahydro Isoquinolin-7-yl)-1H-indazole- 6-carbonitrile

Into a 25 mL round bottom flask equipped with a magnetic stirrer, compound 3-1 (120 mg, 0.17 mmol) and dichloromethane (2.0 mL) were added, and then trifluoroacetic acid (2.0 mL) was added to the reaction solution. The reaction was stirred at room temperature for 2 hours. The oily residue of the solution was concentrated under reduced pressure, anhydrous methanol (3.0 ml) was added, and after cooling to zero in an ice bath, anhydrous potassium carbonate (60 mg) was added to the reaction solution. The reaction was stirred for an additional 20 minutes at room temperature. After the reaction was completed, it was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography to obtain the target product 3 (14.5 mg, yield: 18%). MS(ESI):m/z=470.1[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ8.16(s, 1H), 8.08(s, 1H), 7.59(s, 1H), 7.46(s, 1H), 6.96(d, J=9.2Hz, 1H ),3.99(s,2H),3.70(s,2H),3.20-3.03(m,2H),2.90(t,J＝6.0Hz,2H),2.87-2.80(m,4H),2.48(s, 3H),2.32(s,3H).

The following compounds were obtained using a method similar to that of Example 3, replacing the corresponding raw materials.

Example 14: N-(4-(3-(2,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-6-isocyano-1H-indazole -5-yl)-3-fluoro- 5-methylphenyl)-2-(dimethylamino)acetamide

N-(4-(3-(2,5-Dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-6-isocyano-1-((2-(three Methylsilyl) Ethoxy) Methyl)-1H-indazol-5-yl)-3-fluoro-5-methylphenyl)-2-(dimethylamino)acetamide

Under argon protection, to compound 1-1 (80 mg, 0.17 mmol), C4 (81 mg, 0.24 mmol), methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (15 mg, 0.017 mmol) and potassium phosphate ( 109 mg, 0.51 mmol), 1,4-dioxane (1.5 mL) and water (0.4 mL) were added. Under argon protection, the reaction was heated to 100°C and stirred for 3 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (5 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (dichloromethane/methanol=100:1-15:1) to obtain yellow solid compound 14-1 (56 mg, yield: 51%). MS(ESI):m/z＝641.3[M+H] ⁺ .

N-(4-(3-(2,5-Dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-6-isocyano-1H-indazol-5-yl )-3-fluoro-5-methylbenzene base)-2-(dimethylamino)acetamide

To a solution of compound 14-1 (56 mg, 0.087 mmol) in dichloromethane (2 mL) was added dropwise trifluoroacetic acid (2 mL) under ice-cooling, and the reaction was stirred at room temperature for 2 hours. The solvent was distilled off under reduced pressure to obtain an oily residue, to which anhydrous methanol (3.0 ml) was added. After cooling to zero with an ice-water bath, potassium carbonate (60 mg) was added to the reaction solution and stirred for 20 minutes. After filtration, the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by preparative high performance liquid chromatography to obtain the target product 14 (11 mg, yield: 24.9%). MS(ESI):m/z=511.1[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ8.16(s, 1H), 7.98(s, 1H), 7.62(d, J=2.4Hz, 1H), 7.59(s, 1H), 7.46(s, 1H ),7.35(s,1H),3.69(s,2H),3.18(s,2H),2.91-2.81(m,4H),2.47(s,3H),2.40(s,6H),2.32(s, 3H), 2.14(s, 3H).

The following compounds were obtained using a method similar to that of Example 14, replacing the corresponding raw materials.

Example 16: 5-(6,8-difluoro-1,2,3,4-tetrahydroisoquinolin-7-yl)-3-(2-methyl-1-oxo-1,2,3 ,4-Tetrahydroisoquinoline-7- base)-1H-indazole-6-carbonitrile

7-(5-Chloro-6-isocyano-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-3-yl)-2-methyl- 3,4-dihydro Isoquinolin-1(2H)-one

Under argon protection, compound A1 (200 mg, 0.46 mmol), B5 (159 mg, 0.55 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride To a mixture of methyl chloride complex (38 mg, 0.046 mmol) and potassium phosphate (215 mg, 1.01 mmol), 1,4-dioxane (2.4 mL) and water (0.6 mL) were added. Under the protection of argon, the reaction solution was heated to 90°C and stirred for 15 hours. After filtration, the filtrate was extracted with ethyl acetate (5 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by a reverse silica gel column to obtain the target compound 16-1 (210 mg, yield: 97.8%) as a yellow oil. MS(ESI): m/z=467.2[M+H] ⁺ .

Tert-butyl 7-(6-cyano-3-(2-methyl-1-carbonyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-1-((2-( Trimethylsilyl) B Oxy)methyl)-1H-indazol-5-yl)-6,8-difluoro-3,4-dihydroisoquinoline-2(1H)-carboxylate

Using compound 16-1 (210 mg, 0.45 mmol) and C3 (213 mg, 0.54 mmol) as starting materials, compound 16-2 (259 mg, yield: 82%) was prepared by referring to the synthetic method of compound 14-1. MS(ESI): m/z=722.2[M+Na] ⁺ .

5-(6,8-difluoro-1,2,3,4-tetrahydroisoquinolin-7-yl)-3-(2-methyl-1-carbonyl-1,2,3,4-tetra Hydroisoquinolin-7-yl)-1H-ind Azole-6-carbonitrile

Using compound 16-2 (59 mg, 0.37 mmol) as a raw material, compound 16 (27 mg, yield: 15.5%) was synthesized by referring to the preparation method of Example 14. MS(ESI): m/z=470.2[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD)δ8.51(s,1H),8.24(s,1H),8.20(s,1H),8.09(d,J＝8.0Hz,1H),7.45(d,J =8.0Hz, 1H), 6.96(d, J=9.6Hz, 1H), 4.00(s, 2H), 3.68(t, J=6.0Hz, 2H), 3.18(s, 3H), 3.16-3.07(m ,4H),2.92(m,2H).

The following compounds were obtained using a method similar to that of Example 16, replacing the corresponding raw materials.

Example 18: 3-(2,5-Dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-5-(2-fluoro-6-methyl-4-(piper Pyridine-2-yl)phenyl)- 1H-Indazole-6-carbonitrile

tert-butyl 2-(4-(3-(2,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-6-isocyano-1-(( 2-(trimethylsilyl) Ethoxy)methyl)-1H-indazol-5-yl)-3-fluoro-5-methylphenyl)piperidine-1-carboxylate

Under argon protection, compound 1-1 (110 mg, 0.236 mmol), C5 (120 mg, 0.283 mmol), potassium phosphate (100 mg, 0.471 mmol) and methanesulfonic acid (2-dicyclohexyl Phosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II)( 27 mg, 0.023 mmol), 1,4-dioxane (3.0 mL) and water (1.0 mL) were added. Under the protection of argon, the reaction solution was heated to 90°C and stirred for 14 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (5 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse column chromatography to obtain light yellow solid 18-1 (89 mg, yield: 52.2%). MS(ESI):m/z=724.6[M+H] ⁺ .

3-(2,5-Dimethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-5-(2-fluoro-6-methyl-4-(piperidine-2- Base) phenyl) -1H-indazole-6-carba Nitrile

To a solution of compound 18-1 (89 mg, 0.123 mmol) in dichloromethane (2.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was stirred at room temperature for 2 hours. After complete conversion of starting materials, the reaction solution was concentrated under reduced pressure to obtain an oily residue, and then anhydrous methanol (3.0 mL) was added thereto. The solution was cooled in an ice-water bath, then potassium carbonate (60 mg) was added and stirring was continued for 20 minutes. After filtration, the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography to obtain the title compound 18 (52 mg, yield: 85.7%). MS(ESI): m/z=494.2[M+H] ⁺ .

¹ H NMR (400MHz, MeOD) δ8.24(s, 1H), 8.08(d, J=1.4Hz, 1H), 7.81(s, 1H), 7.65(s, 1H), 7.38(d, J=11.6 Hz,1H),7.29(t,J=11.6,1H),4.71-4.59(m,1H),4.51-4.39(m,1H),4.38-4.30(m,1H),3.91-3.78(m,1H ),3.57-3.46(m,2H),3.29-3.20(m,1H),3.19-3.12(m,2H),3.09(s,3H),2.40(s,3H),2.27-2.14(m,4H ),2.10-2.00(m,3H),1.89-1.76(m,2H).

Example 19: 1-(3-fluoro-4-(6-isocyano-3-(2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-1H-ind Azol-5-yl)-5-carba phenyl)-N-methylmethylamine

5-Chloro-3-(2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-1-((2-(trimethylsilyl)ethoxy)methyl Base) -1H-indazole- 6-carbonitrile

Under argon protection, A1 (130 mg, 0.30 mmol), B8 (90 mg, 0.33 mmol), potassium phosphate (127 mg, 0.06 mmol) and [1,1'-bis(diphenylphosphine ) to a mixture of ferrocene]palladium dichloride (24.5 mg, 0.03 mmol), 1,4-dioxane (2.0 mL) and water (0.6 mL) were added. Under the protection of argon, the reaction solution was placed in a 90 degree oil bath and stirred for 4 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (5 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse silica gel column to obtain compound 19-1 (100 mg, yield: 70.7%). MS(ESI): m/z=453.1,455.1[M+H] ⁺ .

tert-Butyl(3-fluoro-4-(6-isocyano-3-(2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-1-(2-( Trimethylsilyl) B Oxy)methyl)-1H-indazol-5-yl)-5-methylbenzyl)(methyl)carbamate

Under argon protection, compound 19-1 (80 mg, 0.18 mmol), C1 (74 mg, 0.20 mmol), potassium phosphate (112 mg, 0.53 mmol) and methanesulfonic acid (2-dicyclohexyl Phosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II)( 14.4 mg, 0.017 mmol), 1,4-dioxane (1.5 mL) and water (0.5 mL) were added. The reaction solution was heated to 100° C. and stirred for 3 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (5 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse silica gel column to obtain 19-2 (70 mg, yield: 57.2%). MS(ESI):m/z=670.3[M+H] ⁺ .

1-(3-fluoro-4-(6-isocyano-3-(2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-1H-indazole-5- base)-5-methylphenyl)-N- methyl methylamine

Using compound 19-2 (70 mg, 0.104 mmol) as starting material, the target product 19 (10.2 mg, yield: 22.2%) was synthesized by referring to the preparation method of compound 1. MS(ESI): m/z=440.3[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ8.14(s, 1H), 7.97(s, 1H), 7.70(d, J=8.0Hz, 1H), 7.61(s, 1H), 7.27(d, J =8.0Hz,1H),7.17(s,1H),7.12-7.06(m,1H),3.77(s,2H),3.68(s,2H),2.98(t,J=6.4Hz,2H),2.78 (t,J=6.0Hz,2H),2.45(s,3H),2.42(s,3H),2.14(s,3H).

The following compounds were obtained using a method similar to that of Example 19, replacing the corresponding raw materials.

Example 22: 5-(5-amino-4-methylpyridin-3-yl)-3-(6-isopropyl-7-carbonyl-5,6,7,8-tetrahydro-4H-pyrazole and [1,5-d][1,4]diazoheptin-2-yl)-1H-indazole-6-carbonitrile

5-Chloro-3-(6-isopropyl-7-carbonyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-d][1,4]diazoheptin -2-base)-1-((2-(three Methylsilyl)ethoxy)methyl)-1H-indazole-6-carbonitrile

Under argon protection, compound A1 (135 mg, 0.31 mmol), B4 (101 mg, 0.34 mmol), potassium phosphate (132 mg, 0.62 mmol) and [1,1'-bis(diphenyl To a mixture of phosphine)ferrocene]palladium dichloride (25 mg, 0.031 mmol), 1,4-dioxane (2.0 mL) and water (0.5 mL) were added. Under the protection of argon, the reaction solution was heated to 100°C and stirred for 3 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (5 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-1:2) to obtain solid 22-1 (134 mg, yield: 86.2%). MS(ESI):m/z=499.1,501.1[M+H] ⁺ .

tert-butyl (5-(6-isocyano-3-(6-isopropyl-7-oxo-5,6,7,8-tetrahydro-4H-pyrazole-[1,5-d] [1,4]diaza-2- Base)-1-(2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)-4-methylpyridin-3-yl)carbamate

Under argon protection, compound 22-1 (60 mg, 0.12 mmol), 4-N, N-bis-tert-butoxycarbonyl 4-methyl-3-(4,4,5,5-tetramethyl 1,3,2-dioxaborolan-2-yl)pyridine (63 mg, 0.144 mmol), potassium phosphate (51 mg, 0.24 mmol) and methanesulfonic acid (2-dicyclohexylphosphino -2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (10.1 mg , 0.012 mmol), 1,4-dioxane (1.2 ml) and water (0.4 ml) were added. Under the protection of argon, the reaction solution was heated to 90°C and stirred for 3 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (5 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (petroleum ether/ethyl acetate=9:1-1:9) to obtain the title compound 22-2 (56 mg, yield: 59.2%). MS(ESI): m/z=771.4[M+H] ⁺ .

5-(5-amino-4-methylpyridin-3-yl)-3-(6-isopropyl-7-carbonyl-5,6,7,8-tetrahydro-4H-pyrazolo[1, 5-d][1,4]diazo Heptin-2-yl)-1H-indazole-6-carbonitrile

To compound 22-2 (56 mg, 0.072 mmol) in anhydrous dichloromethane (2.0 mL) was added trifluoroacetic acid (1.0 mL) under ice-water bath cooling. The ice bath was removed, and the reaction solution was stirred at room temperature for 2 hours. After complete conversion of starting materials, the reaction solution was concentrated under reduced pressure to obtain an oily residue, and then anhydrous methanol (3.0 mL) was added thereto. The reaction solution was cooled in an ice-water bath, then solid potassium carbonate was added to adjust the pH to 10-11, and stirring was continued for 30 minutes. After filtration, the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography to obtain the target product 22 (8.6 mg, yield: 26.9%) as a white solid. MS(ESI):m/z=441.2[M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ8.32(s,1H),8.07(s,1H),8.03(s,1H),7.79(s,1H),7.75(s,1H),6.68(s, 1H),5.23-5.13(m,2H),4.76-4.70(m,1H),4.51(s,2H),3.89-3.85(m,2H),3.22-3.15(m,2H),2.02(s, 3H), 1.21(d, J=7.2Hz, 6H).

Example 23: 5-(6,8-difluoro-1,2,3,4-tetrahydroisoquinolin-7-yl)-3-(8-methyl-2,3-dihydro-1H- pyrido[2,3- b][1,4]oxazin-7-yl)-1H-indazole-6-carbonitrile

tert-Butyl 7-(5-chloro-6-cyano-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-3-yl)-8- Methyl-2,3- Dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate

Under argon protection, to A1 (300 mg, 0.69 mmol), tert-butyl 8-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborin Alk-2-yl)-2,3-dihydro-1H-pyridyl[2,3-b][1,4]oxazine-1-carboxylate (312.3 mg, 0.83 mmol), [1, To a mixture of 1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (56.7 mg, 0.069 mmol) and potassium phosphate (323 mg, 1.52 mmol), was added 1, 4-Dioxane (1.0 mL) and water (0.3 mL). Under argon protection, the reaction was warmed to 90°C and stirred for 12 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (5 mL×2). The combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography to obtain the title compound 23-1 (283 mg, yield: 73.6%). MS(ESI): m/z=556.2[M+H] ⁺ .

tert-butyl 7-(5-(2-(tert-butoxycarbonyl)-6,8-difluoro-1,2,3,4-tetrahydroisoquinolin-7-yl)-6-cyano- 1-((2-(Triple A ylsilyl)ethoxy)methyl)-1H-indazol-3-yl)-8-methyl-2,3-dihydro-1H-pyridyl[2,3-b][1,4 ]oxazine-1-carboxylate Ester

Under argon protection, to compound 23-1 (140 mg, 0.25 mmol), C3 (120 mg, 0.30 mmol), methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (22 mg, 0.025 mmol) and potassium phosphate ( 161 mg, 0.76 mmol), 1,4-dioxane (2.4 mL)/water (0.6 mL) was added. Under argon protection, the reaction was warmed to 100°C and stirred for 4 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (5 mL×2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography to obtain the title compound 23-2 (160 mg, yield: 80.5%). MS(ESI): m/z=789.4[M+H] ⁺ .

(6,8-Difluoro-1,2,3,4-tetrahydroisoquinolin-7-yl)-3-(8-methyl-2,3-dihydro-1H-pyridyl[2,3 -b][1,4]oxazine-7- base)-1H-indazole-6-carbonitrile

To a solution of compound 23-2 (160 mg, 0.20 mmol) in dichloromethane (3.0 mL) was added trifluoroacetic acid (1.0 mL) dropwise at room temperature. The reaction was stirred at room temperature for 2 hours. The reaction was detected by LCMS. After the reactant disappeared, the solvent was concentrated under reduced pressure. Anhydrous methanol (3.0 ml) was added to the residue, and the reaction solution was cooled to zero with an ice-bath, and potassium carbonate (60 mg) was added thereto, followed by stirring for 20 minutes. After filtration, the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography, and the target product 23 was obtained as a white solid (21.1 mg, yield: 22.3%). MS(ESI): m/z=459.2[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ8.21(s, 1H), 7.67(s, 1H), 7.55(s, 1H), 6.93(d, J=9.6Hz, 1H), 4.46-4.35(m ,2H),3.97(d,J=4.2Hz,2H),3.54-3.44(m,2H),3.17-3.04(m,2H),2.90(s,2H),2.12(s,3H).

Example 24: 7-(6-fluoro-5-(2-fluoro-6-methylphenyl)-1H-indazol-3-yl)-2,5-dimethyl-1,2,3, 4-Tetrahydroisoquinoline

7-(5-Bromo-6-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-3-yl)-2,5-dimethyl -1,2,3,4-four Hydroisoquinoline

Under argon protection, A2 (210 mg, 0.40 mmol, purity: 90%), B1 (127 mg, 0.4411 mmol), potassium phosphate (170 mg, 0.80 mmol) and [1,1'-bis To a mixture of (diphenylphosphine)ferrocene]palladium dichloride (32 mg, 0.04 mmol), 1,4-dioxane (3.0 mL) and water (0.6 mL) were added. After argon replacement three times, the reaction solution was heated to 65°C and stirred for 4 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature, and extracted with ethyl acetate (10 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by a flash reverse silica gel column to obtain the title compound 24-1 (118 mg, yield: 58.3%). MS(ESI):m/z=504.0,506.0[M+H] ⁺ .

7-(6-fluoro-5-(2-fluoro-6-methylphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole-3 -base)-2,5-di Methyl-1,2,3,4-tetrahydroisoquinoline

Under argon protection, compound 24-1 (55 mg, 0.11 mmol), (2-fluoro-6-methylphenyl) boronic acid (20.2 mg, 0.13 mmol), chloro(2-dicyclohexylphosphine yl-2,4,6-triisopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II) (8.6 mg, 0.011 mmol) and To a mixture of potassium phosphate (69.4 mg, 0.33 mmol), dioxane (1.0 mL) and water (0.3 mL) were added. After argon replacement three times, the reaction mixture was warmed up to 100°C and stirred for 3 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature and extracted with ethyl acetate (5 mL×2). The combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by normal phase column chromatography (dichloromethane/methanol=100:1-20:1) to obtain the title compound 24-2 (58 mg, yield: 50.6%). MS(ESI): m/z=534.3[M+H] ⁺ .

7-(6-fluoro-5-(2-fluoro-6-methylphenyl)-1H-indazol-3-yl)-2,5-dimethyl-1,2,3,4-tetrahydro Isoquinoline

To a solution of compound 24-2 (58 mg, 2.59 mmol) in dichloromethane (3.0 mL) was added trifluoroacetic acid (1.0 mL) dropwise at room temperature. The reaction solution was stirred at room temperature for 3 hours. It should be detected by LCMS. After the reactant disappeared, the solvent was concentrated under reduced pressure. Anhydrous methanol (3.0 ml) was added to the residue, the reaction solution was cooled to zero with an ice-water bath, potassium carbonate (70 mg) was added, and the reaction solution was stirred at room temperature for 20 minutes. After filtration and concentration under reduced pressure, the crude product was obtained. The crude product was purified by high performance liquid chromatography to obtain the title compound 24 (11.2 mg, 25.7%) as a white solid. MS(ESI): m/z=404.2[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ7.80(d, J=6.8Hz, 1H), 7.57(s, 1H), 7.44(s, 1H), 7.36(d, J=9.6Hz, 1H), 7.34-7.29(m,1H),7.16(d,J=7.6Hz,1H),7.03(t,J=8.8Hz,1H),3.70(s,2H),2.88-2.80(m,4H),2.48 (s,3H),2.31(s,3H),2.17(s,3H).

The following compounds were obtained using a method similar to that of Example 24, substituting the corresponding raw materials.

Example 26: 1-(3-fluoro-4-(6-fluoro-3-(2-(4-methylpiperazin-1-yl)pyrimidin-5-yl)-1H-indazol-5-yl )-5-Methylbenzene base) -N-methylmethylamine

5-Bromo-6-fluoro-3-(2-(4-methylpiperazin-1-yl)pyrimidin-5-yl)-1-(2-(trimethylsilyl)ethoxy)methyl base)-1H-ind azole

Using compound A2 (120 mg, 0.254 mmol) and (2-(4-methylpiperazin-1-yl) pyrimidin-5-yl) boronic acid (62.0 mg, 0.280 mmol) as raw materials, refer to compound 1-1 Compound 26-1 (90 mg, yield: 67.7%) was synthesized by the method. MS(ESI):m/z=521.1,523.1[M+H] ⁺ .

tert-Butyl(3-fluoro-4-(6-fluoro-3-(2-(4-methylpiperazin-1-yl)pyrimidin-5-yl)-1-(2-(trimethylsilane base) ethoxy) methyl Base)-1H-indazol-5-yl)-5-methylbenzyl)(methyl)carbamate

Using compound 26-1 (90 mg, 0.172 mmol) and C1 (80 mg, 0.207 mmol) as raw materials, 26-2 (56 mg, yield: 46.7%) was synthesized according to the method of compound 1-2. MS(ESI):m/z＝694.3[M+H] ⁺ .

1-(3-fluoro-4-(6-fluoro-3-(2-(4-methylpiperazin-1-yl)pyrimidin-5-yl)-1H-indazol-5-yl)-5- Methylphenyl)-N-methylmethanol amine

Using compound 26-2 (56 mg, 0.081 mmol) as starting material, compound 26 (13 mg, yield: 34.8%) was synthesized according to the method of compound 1. MS(ESI): m/z=464.2[M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.85(s, 2H), 7.77(d, J=6.8Hz, 1H), 7.36(d, J=9.6Hz, 1H), 7.13(s, 1H) ,7.04(d,J＝10.0Hz,1H),3.93-3.86(s,4H),3.75(s,2H),2.55-2.48(m,4H),2.42(s,3H),2.33(s,3H ),2.17(s,3H).

The following compounds were obtained using a method similar to that of Example 26, substituting the corresponding raw materials.

Example 29: 5-(2-(difluoromethoxy)-6-fluorophenyl)-3-(2-(2-hydroxy-2-methylpropyl)-5-methyl-1,2 ,3,4- Tetrahydroisoquinolin-7-yl)-1H-indazole-6-carbonitrile

5-(2-(Difluoromethoxy)-6-fluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole-6-methyl Nitrile

Under argon protection, to compound A1-2 (200 mg, 0.65 mmol), 2-(2-(difluoromethoxy)-6-fluorophenyl)-4,4,5,5-tetramethyl 1,3,2-dioxaborolane (225 mg, 0.78 mmol), potassium phosphate (415 mg, 1.95 mmol) and methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tris-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (56 mg, 0.065 mmol) To the mixture, 1,4-dioxane (3.0 mL) and water (0.6 mL) were added. After argon replacement three times, the reaction solution was heated to 100°C and stirred for 4 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature, and extracted with ethyl acetate (20 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=100:1-7:3) to obtain the title compound 29-1 (214 mg, yield: 75.8%) as light yellow foamy solid. MS(ESI): m/z=434.1[M+H] ⁺ .

5-(2-(Difluoromethoxy)-6-fluorophenyl)-1H-indazole-6-carbonitrile

To compound 29-1 (214 mg, 0.49 mmol) in anhydrous dichloromethane (4.0 mL) was added trifluoroacetic acid (3.0 mL) under ice-cooling. The ice bath was removed, and the reaction was stirred at room temperature for 2 hours. The solution was concentrated under reduced pressure to obtain an oily residue, which was dissolved by adding anhydrous methanol (10 mL). After the solution was cooled to zero with an ice-water bath, anhydrous potassium carbonate was added to the reaction solution to adjust the pH to 10-11. Start the stirrer, stir for 30 minutes, filter, and concentrate the filtrate under reduced pressure to obtain the crude product. The crude product was separated and purified by reverse column chromatography to obtain light yellow foamy solid 29-2 (88 mg, yield: 58.7%). MS(ESI):m/z=304.1[M+H] ⁺ .

5-(2-(Difluoromethoxy)-6-fluorophenyl)-3-iodo-1H-indazole-6-carbonitrile

To a solution of 29-2 (88 mg, 0.29 mmol) in N,N-dimethylformamide (2.0 mL) was added N-iodosuccinimide (72 mg, 0.32 mmol) at room temperature . After the reaction body was stirred at room temperature for 12 hours, the reaction solution was separated and purified by reverse column chromatography to obtain 29-3 (114 mg, yield: 91.6%) as a pale yellow foamy solid. MS(ESI):m/z=429.9[M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ ) δ8.00(s, 1H), 7.61(s, 1H), 7.58-7.42(m, 1H), 7.20-7.06(m, 2H), 6.50(t, J=72.0 Hz,1H).

tert-Butyl 6-cyano-5-(2-(difluoromethoxy)-6-fluorophenyl)-3-iodo-1H-indazole-1-carboxylate

To a solution of compound 29-3 (114 mg, 0.266 mmol) in dichloromethane (2 mL) was added di-tert-butyldicarbonate (116 mg, 0.532 mmol) at room temperature, N,N - Dimethylpyridin-4-amine (3.3 mg, 0.0266 mmol) and triethylamine (54 mg, 0.532 mmol). The reaction solution was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse column chromatography to obtain light yellow foamy solid 29-4 (128 mg, yield: 91.0%). MS(ESI):m/z=429.9[M-56+H] ⁺ .

tert-Butyl 6-cyano-5-(2-(difluoromethoxy)-6-fluorophenyl)-3-(2-(2-hydroxy-2-methylpropyl)-5-methanol base-1,2,3,4- Tetrahydroisoquinolin-7-yl)-1H-indazole-1-carboxylate

Under argon protection, compound 29-4 (115 mg, 0.22 mmol), B10 (99 mg, 0.29 mmol), potassium phosphate (103 mg, 0.48 mmol) and [1,1'-bis(di To a mixture of phenylphosphino)ferrocene]palladium dichloride (18 mg, 0.022 mmol), 1,4-dioxane (5 mL) and water (1 mL) were added. The reaction solution was heated to 70° C. and stirred for 3 hours. The reaction was checked by LCMS. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (25 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse column chromatography to obtain pale yellow foamy solid 29-5 (67 mg, yield: 49.1%). MS(ESI):m/z=621.2[M+H] ⁺ .

5-(2-(Difluoromethoxy)-6-fluorophenyl)-3-(2-(2-hydroxy-2-methylpropyl)-5-methyl-1,2,3,4 -Tetrahydroisoquinolin-7-yl)- 1H-Indazole-6-carbonitrile

To a solution of compound 29-5 (67 mg, 0.108 mmol) in dichloromethane (2 mL) was added trifluoroacetic acid (2 mL) under ice-cooling. After the reaction solution was stirred at room temperature for 1 hour, the reaction was concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography to obtain white solid compound 29 (38.4 mg, yield: 68.5%). MS(ESI): m/z=521.0[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ8.16(s,1H),8.06(s,1H),7.62-7.51(m,2H),7.42(s,1H),7.22-7.18(m,2H) ,6.84(t,J＝72.0Hz,1H),3.86(s,2H),2.99-2.94(m,2H),2.86-2.81(m,2H),2.53(s,2H),2.32(s,3H ),1.24(s,6H).

The following compounds were obtained using a method similar to that of Example 29, substituting the corresponding raw materials.

Example 30: 7-(5-(2-(Difluoromethoxy)-6-fluorophenyl)-6-isocyano-1H-indazol-3-yl)-5-methyl-2- (Methyl-d3)- 1,2,3,4-Tetrahydroisoquinoline

tert-Butyl 7-(5-chloro-6-isocyano-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-3-yl)-5 -methyl- 3,4-Dihydroisoquinoline-2(1H)-carboxylate

Under argon protection, compound A1 (750 mg, 1.73 mmol), B7 (776 mg, 2.08 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (139 mg, 0.17 mmol) and anhydrous potassium phosphate (734 mg, 3.46 mmol) were added 1,4-dioxane (15 mL) and water (3 mL). Under argon protection, the reaction was stirred at 90°C for 6 hours. After the reaction was completed, the reaction solution was concentrated. The residue was purified by normal phase column chromatography (petroleum ether/ethyl acetate=100:1-85:15) to obtain compound 30-1 (300 mg, yield: 31%). MS(ESI):m/z=497.1[M-56+H] ⁺ .

7-(5-Chloro-6-isocyano-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-3-yl)-5-methyl- 1,2,3,4-four Hydroisoquinoline Hydrochloride

Compound 30-1 (300 mg, 0.54 mmol) was dissolved in ethyl acetate (3 ml), the reaction solution was cooled to 0°C, and ethyl acetate hydrochloride solution (5 ml, 4N) was slowly added dropwise to the reaction solution . After the dropwise addition was completed, the reaction solution was warmed up to room temperature, and stirring was continued for 2 hours. After the reaction was completed, the reaction solution was concentrated to obtain the title compound 30-2 (280 mg). MS(ESI): m/z=453.2[M+H] ⁺ .

7-(5-Chloro-6-isocyano-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-3-yl)-5-methyl- 2-(Methyl- d3)-1,2,3,4-Tetrahydroisoquinoline

Deuterated formaldehyde (346 mg, 2.16 mmol, 20% aqueous solution) was added to a solution of 30-2 (280 mg, 0.54 mmol) in methanol (5 mL), and stirred at room temperature for 10 minutes. Then sodium deuterated cyanoborohydride (71 mg, 1.08 mmol) was added to the reaction solution in portions, and stirring was continued at room temperature for 1 hour. The reaction was quenched with saturated sodium bicarbonate solution (20 mL), extracted twice with ethyl acetate (20 mL), the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by reverse phase column chromatography to obtain the title compound 30-3 (180 mg, yield: 71%). MS(ESI): m/z=470.3[M+H] ⁺ .

7-(5-(2-(Difluoromethoxy)-6-fluorophenyl)-6-isocyano-1-((2-(trimethylsilyl)ethoxy)methyl) -1H-indazole -3-yl)-5-methyl-2-(methyl-d3)-1,2,3,4-tetrahydroisoquinoline

Under argon protection, compound 30-3 (180 mg, 0.38 mmol), 2-(2-(difluoromethoxy)-6-fluorophenyl)-4,4,5,5-tetramethyl -1,3,2-dioxaborolane (164 mg, 0.57 mmol), methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1 ,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (32 mg, 0.038 mmol) and anhydrous potassium phosphate (161 mg, 0.76 mmol ), 1,4-dioxane (2 mL) and water (0.5 mL) were added. The reaction mixture was heated to 100° C. and stirred for 4 hours under the protection of argon. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by column chromatography (dichloromethane/methanol=100:1-92:8) to obtain the title compound 30-4 (80 mg, yield: 79%). MS(ESI):m/z=596.2[M+H] ⁺ .

7-(5-(2-(Difluoromethoxy)-6-fluorophenyl)-6-isocyano-1H-indazol-3-yl)-5-methyl-2-(methyl- d3)-1,2,3,4-four Hydroisoquinoline

To a solution of 30-4 (180 mg, 0.30 mmol) in dichloromethane (2 mL) was added trifluoroacetic acid (2 mL). The reaction solution was stirred at room temperature for 2 hours and then concentrated to obtain a residue. The residue was dissolved in methanol (4 mL), cooled to zero degrees Celsius, anhydrous potassium carbonate (208 mg, 1.5 mmol) was added, and the reaction solution was stirred at room temperature for half an hour. After the reaction was completed, the reaction solution was concentrated. The residue was separated and purified by reverse phase column to obtain the title compound 30 (105 mg, yield: 77%). MS(ESI): m/z=466.1[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ8.15(s,1H),8.07(s,1H),7.59(s,1H),7.58-7.52(m,1H),7.46(s,1H),7.23 -7.16(m,2H),6.84(d,J=72.8Hz,1H),3.68(s,2H),2.91-2.79(m,4H),2.31(s,3H).

The following compounds were obtained using a method similar to that of Example 30, substituting the corresponding raw materials.

Biological test example 1 HPK1 ADP-Glo enzymatic activity test

Prepare enzyme activity assay buffer containing 40mM Tris, pH 7.5; 20mM MgCl ₂ ; 0.1 mg/mL BSA; 50 μM DTT. Compounds were dissolved in 100% DMSO, stock solution concentration was 10 mM. The DMSO solution of the compound was serially diluted three times from 100 μM, with a total of eleven concentrations. After the diluted compound was diluted 1:20 with the enzyme activity test buffer, 1 μL was added to the working well, and each concentration was two times. Repeat hole. Add 1 μL of 1:20 diluted DMSO solution to negative control wells and positive control wells. Prepare 2.5× substrate/ATP working solution as enzymatic activity test buffer containing 0.25 μg/μL MBP protein and 45 μM ATP, and add 2 μL 2.5× substrate/ATP working solution to each working well. Prepare 2.5×enzyme reaction working solution as enzyme activity test buffer containing 0.5ng/μL HPK1 recombinant protein (Signalchem, Cat. No. M23-11G-10), add 2μL 2.5×enzyme reaction working solution to each working well, negative control well Add only 2 µL of Enzyme Assay Buffer. Attach the sealing film and centrifuge at room temperature for 30 minutes. After the reaction, add 5 μL ADP-Glo reagent (Promega, product number V1901) to each well and react for 40 minutes at room temperature, then add 10 μL kinase detection reagent (Promega, product number V1901). After reacting at room temperature for 20 minutes, the final luminescent signal was measured.

The average values of positive wells and negative wells were calculated as positive control values (Signal _pos ) and negative control values (Signal _neg ), respectively. The signal value of the working well (Signal _test ) was calculated according to the formula Inhibition rate=(Signal _pos -Signal _test )/(Signal _pos -Signal _neg )×100% to calculate the inhibition rate. The obtained inhibition rate was drawn in the GraphPad Prism software according to nonlinear fitting to draw the concentration-inhibition rate curve, and the IC ₅₀ was calculated.

Biological test example 2 GLK ADP-Glo enzyme activity test

Prepare enzyme activity assay buffer containing 40mM Tris, pH 7.5; 20mM MgCl ₂ ; 0.1 mg/mL BSA; 50 μM DTT. Compounds were dissolved in 100% DMSO, stock solution concentration was 10 mM. The DMSO solution of the compound was serially diluted three-fold from 1 mM, with a total of eleven concentrations. After the diluted compound was diluted 1:20 with the enzyme activity test buffer, 1 μL was added to the working well, and each concentration was two times. Repeat hole. Add 1 μL of 1:20 diluted DMSO solution to negative control wells and positive control wells. Prepare 2.5×substrate/ATP working solution as enzymatic activity test buffer containing 0.5 μg/μL PKA substrate polypeptide and 105 μM ATP, and add 2 μL 2.5× substrate/ATP working solution to each working well. Prepare 2.5×enzyme reaction working solution as enzyme activity test buffer containing 2.5ng/μL GLK recombinant protein (Signalchem, Cat. No. M25-11G-10), add 2μL 2.5×enzyme reaction working solution to each working well, negative control well Add only 2 µL of Enzyme Assay Buffer. Attach the sealing film and centrifuge briefly at room temperature for 1 hour reaction. After the reaction, add 5 μL ADP-Glo reagent (Promega, product number V1901) to each well and react at room temperature for 40 minutes, then add 10 μL kinase detection reagent (Promega, product number V1901). After reacting at room temperature for 20 minutes, the final luminescent signal was measured.

The average values of positive wells and negative wells were calculated as positive control values (Signal _pos ) and negative control values (Signal _neg ), respectively. The signal value of the working well (Signal _test ) was calculated according to the formula Inhibition rate=(Signal _pos -Signal _test )/(Signal _pos -Signal _neg )×100% to calculate the inhibition rate. The obtained inhibition rate was drawn in the GraphPad Prism software according to nonlinear fitting to draw the concentration-inhibition rate curve, and the IC ₅₀ was calculated. The experimental results are shown in the table below:

Table 1 Results of HPK1 and GLK enzyme activity experiments

实施例编号Example number	HPK1 ADP-Glo IC50(nM)HPK1 ADP-Glo IC50(nM)	GLK ADP-Glo IC50(nM)GLK ADP-Glo IC50(nM)
11	AA	AA
22	AA	BB
33	AA	AA
44	AA	BB
55	AA	AA
66	AA	AA
77	BB	BB
88	AA	AA
99	AA	AA
1010	AA	BB
1111	AA	AA
1212	AA	BB
1313	AA	AA
1414	AA	AA
1515	AA	AA
1616	AA	AA
1717	AA	AA
1818	AA	AA
1919	AA	AA
2020	AA	AA
21twenty one	AA	AA
22twenty two	BB	BB
23twenty three	BB	BB
24twenty four	BB	BB
2525	AA	AA
2626	AA	AA
2727	AA	AA
2828	BB	AA
2929	AA	CC
3030	AA	AA
3131	BB	CC
3232	BB	CC
3333	AA	AA
3434	AA	CC
3535	BB	BB

3636	AA	BB
3737	BB	CC
3838	BB	CC

Among them, A means IC ₅₀ value≤50nM; B means 50nM<IC ₅₀ value≤500nM; C means 500nM<IC ₅₀ value≤10uM.

Biological test example 3 SLP76 phosphorylation cytology test

Inhibition of HPK1 can inhibit the phosphorylation of its downstream SLP76. Phosphorylation of SLP76 protein was performed using Jurkat (ATCC, Clone E6-1

TIB-152 ^TM ) cells were tested. On the first day of the experiment, the cells were diluted to 10 ⁶ /mL with medium (RPMI 1640+0.5% FBS), and the amount of 10 ⁵ cells per well was spread in 96-well cell culture Plates were starved for 4 hours for incubation. Compounds were dissolved in 100% DMSO with a stock solution concentration of 4 mM. The DMSO solution of the compound was serially diluted four times from 4 mM, with a total of 9 concentrations. Dilute 4 μL of the diluted compound to 196 μL of 37-degree preheated RPMI 1640 and mix well. Add 50 μL of the final diluted compound to the cells, incubate at 37 degrees for 20 minutes, add 50 μL of diluted human CD3/CD28 T cell activator (Stemcell, product number: 10971), so that the final concentration of the activator volume is 1% of the total system. /40, incubate at 37 degrees for 30 minutes. After the reaction, the cells were placed in a 4-degree centrifuge at 1200 rpm for 5 minutes, the medium was sucked away, and 150 μL of cell lysate (provided in the ELISA kit, Cell Signaling, product number 30794C) was added and placed on ice for 30 minutes to fully lyse the cells. After the cells were pipetted evenly, they were placed in a 4-degree centrifuge and centrifuged at 4000 rpm for 5 minutes, and then 50 μL of the supernatant was added to the FastScan ^TM Phospho-SLP-76 (Ser376) ELISA Kit (Cell Signaling, Cat. No. 30794C) to test the phosphorylation level of SLP76 in the cells .

Table 2 SLP76 phosphorylation inhibition experiment results

实施例编号Example number	IC50(nM)IC50(nM)
11	AA
22	BB
33	AA
44	AA
55	AA
66	BB
88	AA
99	AA
1010	AA
1111	BB
1212	BB
1313	AA
1414	AA
1515	AA
1616	BB
1717	AA
1818	AA
1919	AA
2020	AA

21twenty one	AA
23twenty three	BB
2525	BB
2626	AA
2727	AA
3030	AA
3131	BB
3232	BB
3333	AA
3434	BB
3535	BB
3636	BB
3838	BB

Among them, A means IC ₅₀ value ≤ 1000nM; B means 1000nM < IC ₅₀ value ≤ 20000nM;

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims

A compound represented by the following formula I, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof;

in,

- M 2 , M 3 - each independently CR 1 ;-

R is selected from the group consisting of CN, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, phenyl, substituted or unsubstituted C3-C8 cycloalkyl, or -CONH2 ;

A - selected from the following group:
Wherein, V 1 , V 2 , V 3 , V 4 , V 5 , V 6 , V 7 or V 8 are each independently CR 2 or N or NR 2 ;

Each R 2 is independently selected from the group consisting of H, CN, halogen, NH 2 , substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, phenyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group with 1-3 heteroatoms selected from N, S and O in the following group, -C(O)-(substituted or unsubstituted A 3-12-membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O), -(CH 2 ) m N(R 3 ) 2 , -C(O)(CH 2 ) m N(R 3 ) 2 , -NHC(O)(CH 2 ) m N(R 3 ) 2 ; wherein each of said R 3 is independently selected from the following group: H, substituted or unsubstituted C1-C6 Alkyl group, or two R 3 together with the connected N atoms form a 4-7 membered nitrogen-containing heterocyclic ring;

Or two R2 located on adjacent ring atoms together form a 5-7 membered substituted or unsubstituted cyclic group;

m is 0, 1, 2, 3, 4 or 5;

Ra has the structure shown in the following formula:

Wherein, the ring B is a substituted or unsubstituted C3-C8 cycloalkyl group, a substituted or unsubstituted 6-10 membered aryl group, a substituted or unsubstituted 5-12 membered heteroaryl group, a substituted or unsubstituted A 3-15 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O; and the ring B is a bicyclic structure;

n is 0, 1, 2, 3, 4 or 5;

Each R is independently selected from the group consisting of H, halogen, CN, oxygen atom (=O), substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted Substituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O, substituted or unsubstituted C6-C10 aryl , a substituted or unsubstituted 3-10 membered heteroaryl group having 1-3 heteroatoms selected from the group consisting of N, S and O, R 7 -C(=O)-, R 7 -C(=O) -3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O-, R 8 -S(=O) 2 -, R 9 R 10 NC(=O)-, R 9 R 10 NC(=O)NR-, R 11 R 12 NS(=O) 2 -, R 7 O-, R 8 -S(O) 2 NR-; and two R 4 can together form a substituted or unsubstituted 5-7 membered carbocyclic or heterocyclic ring (including saturated or unsaturated ring);

Each R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , and R are each independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkane Oxygen, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 6-10 membered aryl, substituted or unsubstituted with 1-3 heteroatoms selected from the group consisting of N, S and O 3-12 membered heterocyclic group; or said R 9 and R 10 or R 11 and R 12 together form a substituted or unsubstituted nitrogen atom with 1-3 heteroatoms selected from N, S and O The 3-12 membered heterocyclic group;

Unless otherwise specified, the "substitution" refers to being substituted by one or more (such as 2, 3, 4, etc.) substituents selected from the following group: D, halogen, hydroxyl, C1-C6 alkoxy Group, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, methylsulfone, oxo (=O), -CN, -NH 2 , -NHS (O) 2 CH 3 , C1-C6 amino group, carboxyl group, C1-C6 amido group (-C(=O)-N(Rd) 2 or -NH-C(=O)(Rd), Rd is H or C1-C5 alkyl), or unsubstituted or substituted by one or more substituents selected from Z group, selected from the following groups: C1-C6 alkyl, C6-C10 aryl, with 1-3 5-10 membered heteroaryl, -(CH 2 )-C6-C10 aryl, -(CH 2 )-(having 1-3 heteroatoms selected from N, S and O 3-10 membered heteroaryl group with heteroatoms), -(5-10 membered heteroarylene group with 1-3 heteroatoms selected from N, S and O)-(C1-C6 alkyl), with 1-3 3-12-membered heterocyclic groups (including monocyclic, spiro, bridged or parallel rings) with 1-3 heteroatoms selected from the following group of N, S and O, and the group Z substituents are selected from the following Groups: halogen, C1-C6 alkyl, C1-C6 alkylene-OH, C1-C6 alkoxy, oxo, -S(O) 2 CH 3 , -CN, -OH, C6-C10 aryl, A 3-10 membered heterocyclic group having 1-3 heteroatoms selected from N, S and O, -C(O)CH 2 NH 2 , -C(O)CH 2 OH;

And in the compound of formula I, each chiral center is in R configuration or S configuration.
The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein the B ring is selected from the group consisting of 5-6 membered aryl (5- 7-membered partially unsaturated or aromatic heterocycle), 5-7-membered heteroaryl (5-7-membered partially unsaturated or aromatic heterocycle), with 1-3 members selected from the group consisting of N, S and O 3-10 membered heterocyclic group of heteroatoms and (5-7 membered partially unsaturated or aromatic heterocyclic ring), [3-8 membered saturated or partially unsaturated ring] spiro [3-8 membered saturated or partially Unsaturated ring] (can be carbocyclic or heterocyclic), or 6-7 membered aryl or 5-7 membered heteroaryl substituted by at least one substituent selected from the following group: substituted or unsubstituted C3-C8 Cycloalkyl, substituted or unsubstituted 6-10 membered aryl, substituted or unsubstituted 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O.
The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein the ring B is a substituted or unsubstituted group selected from the group consisting of:

Wherein, said U 1 , U 2 , U 3 , U 4 , U 5 , U 6 , U 7 , and U 8 are each independently selected from the following group: CR 1 or N;

Said Y 1 , Y 2 , Y 3 , Y 4 or Y 5 are each independently selected from the following group: None, or C(R 5 ) 2 , C(R 5 ) 2 C(R 5 ) 2 , NR 5 , O or S;

Each of J and K is independently selected from the group consisting of none, or C(R 6 ) 2 , NR 6 , O or S;

Wherein, p is 0, 1, 2 or 3;

q is 0, 1, 2 or 3;

Each R 5 is independently selected from the group consisting of H, CN, halogen, NH 2 , substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, phenyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group (preferably 3-6 membered heterocyclic group) having 1-3 heteroatoms selected from N, S and O in the following group, -(CH 2 ) m N(R 3 ) 2 , -C(O)N(R 3 ) 2 , -NHC(O)(CH 2 ) m N(R 3 ) 2 ; wherein, each of the R 3 Independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, or two R 3 together with the connected N atoms form a 4-7 membered nitrogen-containing heterocyclic ring;

Or two R 5 jointly constitute a group selected from the group consisting of: =O, -(J) n -;

Or R5 located on two adjacent ring atoms and the connected ring atoms together form a substituted or unsubstituted 5-7 membered partially unsaturated or aromatic heterocyclic ring;

Each R 6 is independently selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl;

The linking site can be located on any ring atom of U 1 , U 2 , U 3 , U 4 , U 5 , U 6 , U 7 , U 8 , Y 1 , Y 2 , Y 3 , Y 4 , J and K .
The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein the ring B is a substituted or unsubstituted group selected from the group consisting of: 6 -7-membered aryl (5-7 membered partially unsaturated or aromatic heterocycle), 5-7 membered heteroaryl (5-7 membered partially unsaturated or aromatic heterocycle).
The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein said M3 is CR b ; and said R b is selected from the group consisting of: CN, halogen, -C(O)N(R 6 ) 2 , substituted or unsubstituted C1-C6 alkyl.
The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein the A ring is selected from the group consisting of:
Wherein, V 1 , V 2 , V 3 , V 4 , V 5 , V 6 or V 7 are each independently CR 2 or N; V 8 is NR 2 ; each R 2 is independently selected from the following group: H, CN, halogen, NH 2 , substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, phenyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted A 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group N, S and O, -C(O)-substituted or unsubstituted having 1-3 heteroatoms selected from the group N, S and O 3-12 membered heterocyclic group with O heteroatom, -(CH 2 ) m N(R 3 ) 2 , -C(O)(CH 2 ) m N(R 3 ) 2 , -NHC(O)(CH 2 ) m N(R 3 ) 2 ; wherein, each of said R 3 is independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, or two R 3 together with the connected N atom 4-7 membered nitrogen-containing heterocyclic ring.
The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein the ring A is selected from the group consisting of:
Wherein, said R 2 are each independently selected from the following group: H, CN, halogen, unsubstituted or halogenated C1-C4 alkyl, unsubstituted or halogenated C1-C4 alkoxy;

And said R c is selected from the following group: substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered with 1-3 heteroatoms selected from the group N, S and O Heterocyclic group, -C(O)-substituted or unsubstituted 3-12 membered heterocyclic group with 1-3 heteroatoms selected from the group consisting of N, S and O, -(CH 2 ) m N(R 3 ) 2 , -C(O)(CH 2 ) m N(R 3 ) 2 , -NHC(O)(CH 2 ) m N(R 3 ) 2 ; wherein, the R 3 are each independently selected from The following group: H, substituted or unsubstituted C1-C6 alkyl, or two R 3 together with the connected N atoms form a 4-7 membered nitrogen-containing heterocyclic ring.
The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein the compound of formula I has the structure shown in the following formula II-1 or II-2 :

in,

The R b are each independently selected from the group consisting of halogen, CN, substituted or unsubstituted C1-C6 alkyl.
The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein the compound of formula I is selected from the group consisting of:
The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein the compound of formula I is selected from the group consisting of:
A kind of pharmaceutical composition, it is characterized in that, described pharmaceutical composition comprises (1) compound as claimed in claim 1 or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, Hydrate or solvate; (2) pharmaceutically acceptable carrier.
The compound as claimed in claim 1 or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate, or the purposes of the pharmaceutical composition as claimed in claim 11, It is characterized in that it is used for preparing a pharmaceutical composition for preventing and/or treating diseases related to the activity or expression of HPK1 kinase.
The use according to claim 12, wherein the disease is selected from the group consisting of lymphoma, blastoma, medulloblastoma, retinoblastoma, sarcoma, liposarcoma, synovial cell sarcoma, neuropathic Endocrine tumor, carcinoid tumor, gastrinoma, islet cell carcinoma, mesothelioma, schwannoma, acoustic neuroma, meningioma, adenocarcinoma, melanoma, leukemia or lymphoid malignancy, squamous cell carcinoma, epithelial squamous Stem cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, adenocarcinoma lung cancer, lung squamous cell carcinoma, peritoneal cancer, hepatocellular carcinoma, gastric cancer, bowel cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer , liver cancer, bladder cancer, breast cancer, metastatic breast cancer, colon cancer, rectal cancer, colorectal cancer, uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, anal cancer, penile cancer, Meck Cell carcinoma, esophageal cancer, biliary tract neoplasm, head and neck cancer, hematologic malignancies, nasopharyngeal carcinoma, multiple myeloma, villous adenoma of large field, non-Hodgkin's lymphoma, bone cancer, testicular cancer, Hodgkin's seminoma, oral cavity cancer, brain cancer, skin cancer, breast ductal carcinoma, renal pelvis carcinoma, Wilms tumor, esophageal adenoma, retinoblastoma, glioma, neurofibroma, gastrointestinal stromal tumor tumor, carcinoma in situ, endometrial carcinoma and myelodysplastic syndrome.
The combined tumor chemotherapy regimen of the compound as claimed in claim 1 and its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate and deuterated compound and other tumor immunotherapeutic agents, said other tumors The immunotherapeutic agent is selected from the following group: small molecule compounds and antibodies (including but not limited to PD-1, PD-L1, CTLA-4, TIM-3, TGF-β and its receptors, LAG3 antagonists or TLR4, TLR7, TLR8, TLR9, STING agonists, etc.), radiotherapy regimens, tumor-targeted drugs, and tumor vaccines.
The application of the compound as claimed in claim 1 and pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates and deuterated compounds in combination with CAR-T immunotherapy in cancer immunotherapy.