Tyrosine threonine kinase pkmyt1 inhibitor, and preparation method therefor and use thereof

Abstract

Disclosed in the present invention are a tyrosine threonine kinase PKMYT1 inhibitor, and a preparation method therefor and the use thereof. The inhibitor compound can inhibit the excessive activation of CDK1 in CCNE1-amplified cells by means of inhibiting PKMYT1 kinase, and trigger mitosis prior to the completion of DNA replication, thereby resulting in chromosome pulverization, catastrophic DNA damage and cell death. Therefore, the inhibitor compound can be used for the prevention and / or treatment of related diseases such as cancers (especially endometrial cancer, ovarian cancer, non-small cell lung cancer, esophagogastric junction cancer, etc.), can fill the gap in targeted drugs for cancers of the specific CCNE1-amplified subtype, and has a very good application prospect and value in the field of medicine.

Description

A tyrosine threonine kinase PKMYT1 inhibitor, its preparation method and application Technical Field

[0001] This invention relates to the field of chemical medicine technology, specifically to a tyrosine threonine kinase PKMYT1 inhibitor, its preparation method and application, particularly in the treatment of tumors (especially cancers with CCNE1 gene amplification, including endometrial cancer, ovarian cancer, non-small cell lung cancer, esophageal and gastric cancer, etc.). Background Technology

[0002] Overexpression of CCNE1 deregulated the cell cycle in the G1 / S and G2 / M phases by accelerating S-phase entry, influencing centrosome expansion, and stimulating accelerated mitotic processes. CCNE1 gene amplification is common in various types of tumors, especially in high-grade serous ovarian cancer, uterine tumors, and gastroesophageal tumors. High expression of CCNE1 is associated with genomic instability, genome-wide proliferation, cytotoxicity, and resistance to targeted therapy. [Diagnostics, 2020, 10(5):279.]

[0003] In 2022, a study reported genome-wide CRISPR-Cas9-based synthetic lethality screening in a CCNE1-amplified cell model, showing that amplified CCNE1 leads to increased cellular sensitivity to inhibition of PKMYT1 kinase (a negative regulator of CDK1). Treatment with PKMYT1 kinase inhibitors results in selective and unintended activation of CDK1 in CCNE1-overexpressing cells, promoting early mitosis in cells undergoing DNA synthesis. CCNE1 overexpression disrupts CDK1 homeostasis at least in part through early activation of the MMB-FOXM1 mitotic transcriptional program. Therefore, PKMYT1 inhibition is a promising therapeutic strategy for CCNE1-amplified cancers [Nature, 2022, 604(7907):749-756.].

[0004] Oncology drug discovery based on the recognition of synthetic lethal interactions holds great promise, but to date, few candidate drugs have been developed from scratch using this approach. By developing novel PKMYT1 kinase inhibitors, it is possible to selectively inhibit the growth of CCEN1-amplified cancer cells while exhibiting good safety in CCEN1-normal adjacent tissues. Therefore, PKMYT1 kinase inhibitors have the potential to become precision medicines for cancer patients characterized by CCEN1 amplification. Summary of the Invention

[0005] To overcome the shortcomings of the prior art, the present invention provides a tyrosine threonine kinase PKMYT1 inhibitor, its preparation method and application.

[0006] In a first aspect of the present invention, a compound and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, and solvates are provided, said compound having the following structure:

[0007] in,

[0008] X is selected from: N, CH;

[0009] Ring A is a 3-15 membered carbon ring or a heterocyclic ring;

[0010] R1, R2, R3, R4, R1', R2', R3', and R4' are independently selected from: H, D, halogen, cyano, nitro, and C1-C. 10 Alkyl, C3-C 10 cycloalkyl, C1-C 10 Haloalkyl, -O(C) 0-10 alkyl), -N(C) 0-10 Alkyl)(C 0-10 alkyl);

[0011] R a It is one or more independent substituents on ring A, selected from: H, D, =O, halogen, cyano, nitro, C1-C. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl group, -L0-(C3-C 10 cycloalkyl), -L0-(C6-C 10 aryl), -L0-(4-10 membered heterocyclic group), -N(C 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl) (C3-C 10 cycloalkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -O (C3-C) 10 cycloalkyl), -S(C 0-10 Alkyl), -S(C3-C 10 cycloalkyl), -SO(C 0-10 alkyl), -SO2(C 0-10 Alkyl group), -SO2 (C3-C) 10 cycloalkyl), -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -SO2N(C0-10 Alkyl) (C3-C 10 cycloalkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 Alkyl), -CON(C) 0-10 Alkyl) (C3-C 10 cycloalkyl), -CO(C 0-10 alkyl), -Si(C) 0-10 Alkyl)(C 0-10 Alkyl)(C 0-10 Alkyl); wherein the C0-C6 alkylene, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl group, C3-C 10 cycloalkyl, C6-C 10 The hydrogen atom on the aryl or 4-10 membered heterocyclic group is optionally substituted by a group selected from the following: =O, halogen, cyano, nitro, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C3-C6 alkylene) 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl) (C3-C 10 cycloalkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -O (C3-C) 10 cycloalkyl), -S(C 0-10 Alkyl), -S(C3-C 10 cycloalkyl), -SO(C 0-10 alkyl), -SO2(C 0-10 Alkyl group), -SO2 (C3-C) 10 cycloalkyl), -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -SO2N(C 0-10 Alkyl) (C3-C10 cycloalkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 Alkyl), -CON(C) 0-10 Alkyl) (C3-C 10 cycloalkyl), -CO(C 0-10 alkyl), -Si(C) 0-10 Alkyl)(C 0-10 Alkyl)(C 0-10 alkyl);

[0012] L0 is selected from: single bond, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkyneylene, C3-C7 cycloalkylene, O, S, NH, CONH, NHCO, C(O); wherein the H on the C1-C6 alkylene and C3-C7 cycloalkylene groups is optionally substituted by a group selected from: halogen, cyano, hydroxyl, amino.

[0013] In some embodiments of the present invention, X is CH; in other embodiments of the present invention, X is N.

[0014] In some embodiments of the present invention, ring A is a 3-8 member aliphatic ring, for example...

[0015] In some embodiments of the present invention, ring A is an aromatic ring, for example...

[0016] In some embodiments of the present invention, ring A is a 4-12 membered heteroaromatic ring, for example:

[0017] In some embodiments of the present invention, ring A is a 3-12 member saturated or partially unsaturated heterocycle, for example:

[0018] In some embodiments of the present invention Some have the following structure:

[0019] In some embodiments of the present invention Some have the following structure:

[0020] In some embodiments of the present invention Some have the following structure:

[0021] In some embodiments of the present invention, L0 is a single bond.

[0022] In some embodiments of the present invention, each R a It can be independently selected from: H, =O, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, phenyl, 4-10 membered heterocyclic group, -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 alkyl), -N(C) 0-6 Alkyl)CO(C 0-6 Alkyl), -CON(C) 0-6 Alkyl)(C 0-6 Alkyl), -CO(C) 0-6 Alkyl), -COO(C 0-6 Alkyl), -OCO(C 1-6 Alkyl), -SO(C) 1-6 alkyl), -SO2(C 1-6 Alkyl); wherein the H on the C1-C6 alkyl, C3-C7 cycloalkyl, phenyl, or 4-10 membered heterocyclic group is optionally substituted by a group selected from the following groups: halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 alkyl), -N(C) 0-6 Alkyl)CO(C 0-6 alkyl), -CO(C) 0-6 Alkyl), -CON(C) 0-6 Alkyl)(C 0-6 alkyl), -COO(C 0-6 Alkyl), -OCO(C 1-6 Alkyl), -SO(C) 1-6 alkyl), -SO2(C 1-6 alkyl).

[0023] More specifically, each R a It can be independently selected from: H, =O, F, Cl, Br, I, CN, methyl, ethyl, isopropyl, -CF3, -CHF2, -CH2F, -OH、

[0024] In some embodiments of the present invention, R a It has the following structure: R5 is selected from: C1-C6 alkyl, C3-C7 cycloalkyl, phenyl, 4-8 membered saturated heterocyclic groups; wherein the H on the alkyl, cycloalkyl, phenyl, or saturated heterocyclic group is optionally substituted by a group selected from: -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 Alkyl groups (e.g., -OH, ...) -NH2、 ).

[0025] More specifically, each R a Can be selected independently from:

[0026] In some embodiments of the present invention, R a It has the following structure: Wherein, L1 is selected from: C0-C6 alkylene, C3-C7 cycloalkylene, and R6 is selected from: H, halogen, cyano, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, phenyl, 4-10 membered heterocyclic group, -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 alkyl), -N(C) 0-6 Alkyl)CO(C 0-6 Alkyl), -CON(C) 0-6 Alkyl)(C 0-6 alkyl), -CO(C) 0-6 alkyl), -COO(C 0-6 Alkyl), -OCO(C 1-6 Alkyl), -SO(C) 1-6 alkyl), -SO2(C 1-6 Alkyl); wherein the H on the C1-C6 alkyl, C3-C7 cycloalkyl, phenyl, or 4-10 membered heterocyclic group is optionally substituted by a group selected from the following groups: halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 alkyl), -N(C) 0-6 Alkyl)CO(C 0-6 alkyl), -CO(C) 0-6 Alkyl), -CON(C) 0-6 Alkyl)(C 0-6 alkyl), -COO(C 0-6 Alkyl), -OCO(C 1-6 Alkyl), -SO(C) 1-6 alkyl), -SO2(C 1-6alkyl).

[0027] More specifically, each R a Can be selected independently from:

[0028] In some embodiments of the present invention, R a It has the following structure: -L2-R7, where L2 is selected from: NH, CONH, NHCO, C(O); R7 is selected from: C3-C7 cycloalkyl, phenyl, 4-10 membered heterocyclic group, wherein the H on the cycloalkyl, phenyl, or 4-10 membered heterocyclic group is optionally substituted by a group selected from: halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 alkyl), -N(C) 0-6 Alkyl)CO(C 0-6 alkyl), -CO(C) 0-6 Alkyl), -CON(C) 0-6 Alkyl)(C 0-6 alkyl), -COO(C 0-6 Alkyl), -OCO(C 1-6 Alkyl), -SO(C) 1-6 alkyl), -SO2(C 1-6 alkyl).

[0029] More specifically, each R a Can be selected independently from:

[0030] Specifically, R1 is selected from: H, halogen, cyano, C1-C6 alkyl, C3-C7 cycloalkyl, C1-C6 haloalkyl, and C1-C6 alkoxy. In some embodiments of the present invention, R1 is selected from: H and halogen (such as F).

[0031] Specifically, R2 is selected from: H, halogen. In some embodiments of the present invention, R2 is H.

[0032] Specifically, R3 is selected from: H, halogens, and C1-C6 alkyl groups. In some embodiments of the present invention, R3 is H.

[0033] Specifically, R4 is selected from: H, halogens, and C1-C6 alkyl groups. In some embodiments of the present invention, R4 is H.

[0034] In some embodiments of the present invention, the compound has the following structure:

[0035] Specifically, R1', R2', R3', and R4' are independently selected from: H, halogen, cyano, C1-C6 alkyl, C3-C7 cycloalkyl, C1-C6 haloalkyl, and C1-C6 alkoxy.

[0036] Specifically, R1' is selected from: H, C1-C6 alkyl; in some embodiments of the present invention, R1' is H. In some embodiments of the present invention, R1' is C1-C3 alkyl, such as methyl.

[0037] Specifically, R2' is selected from: H, C1-C6 alkyl; in some embodiments of the present invention, R2' is H.

[0038] Specifically, R3' is selected from: H, C1-C6 alkyl; in some embodiments of the present invention, R3' is C1-C3 alkyl, such as methyl.

[0039] Specifically, R4' is selected from: H, C1-C6 alkyl; in some embodiments of the present invention, R4' is C1-C3 alkyl, such as methyl.

[0040] In some embodiments of the present invention, the compound has the following structure:

[0041] In some embodiments of the present invention, the compound has the following structure:

[0042] In some embodiments of the present invention, the compound has the following structure:

[0043] In a second aspect of the invention, a method for preparing the compound described in the first aspect is provided, comprising the following steps:

[0044] (1) Compound 1, Compound 2, catalyst, base and solvent are mixed and reacted to obtain Compound 3;

[0045] (2) Compound 3, compound 4, catalyst, base and solvent are mixed and reacted to obtain compound 5;

[0046] (3) Compound 5 was mixed with a carbonylating agent and reacted to obtain compound 6;

[0047] or,

[0048] (1) Compound 1, Compound 2, catalyst, base and solvent are mixed and reacted to obtain Compound 3;

[0049] (2) Compound 3, compound 4', catalyst, base and solvent are mixed and reacted to obtain compound 5';

[0050] (3) Compound 5' was mixed with a carbonylating agent and reacted to obtain compound 6';

[0051] in,

[0052] R P1 It is an alkyl group (such as methyl);

[0053] R P2 It is a halogen or a thiol group;

[0054] R P3 It is a halogen or a thiol group;

[0055] R P4 It is a boric acid group or a borate ester group;

[0056] R P5 It is a boric acid group or a borate ester group;

[0057] R P6 It is protected by H or an amino group;

[0058] R P5 'It is a boric acid group or a borate ester group;

[0059] R P6 ' is a protecting group consisting of H or hydroxyl groups.

[0060] Furthermore, based on the reactivity of the functional groups and the coupling sequence, those skilled in the art can rationally select R. P2 R P3 The choice of the group, for example, in some embodiments of the invention, R P2 For Br, R P3 It is Cl.

[0061] In some embodiments of the present invention, R P4 It is a borate ester group, such as

[0062] In some embodiments of the present invention, R P5 It is a borate ester group, such as

[0063] In some embodiments of the present invention, R P6 H is present, and compound 6 is the target product; in some embodiments of the present invention, R... P6 The amino protecting group is used, and if necessary, the preparation method further includes a deprotection step.

[0064] Specifically, the molar ratio of compound 1 to compound 2 in step (1) is 1:1-2 (e.g., 1:1.1, 1:1.2, 1:1.4, 1:1.5).

[0065] Specifically, the catalyst in step (1) is a zero-valent palladium complex, such as tetra(triphenylphosphine)palladium.

[0066] Specifically, the molar ratio of compound 1 to catalyst in step (1) is 1:0.01-0.1 (e.g., 1:0.02, 1:0.04, 1:0.05, 1:0.06, 1:0.08).

[0067] Specifically, the alkali mentioned in step (1) is selected from sodium carbonate, potassium carbonate, lithium carbonate and cesium carbonate, such as potassium carbonate.

[0068] Specifically, the molar ratio of compound 1 to the base in step (1) is 1:1.5-3 (e.g., 1:1.8, 1:2, 1:2.2, 1:2.5).

[0069] Specifically, the solvent in step (1) can be a mixture of 1,4-dioxane and water (e.g., 4:1, v / v).

[0070] Specifically, the reaction temperature in step (1) is 60-100℃ (e.g., 60, 70, 80, 85, 90, 95, 100℃), for example, 80-95℃.

[0071] Specifically, the reaction time in step (1) is 1-12 hours (e.g., 2, 3, 4, 5, 6, 8, 10, 12 hours).

[0072] Specifically, the molar ratio of compound 3 to compound 4 in step (2) is 1:1-2 (e.g., 1:1.1, 1:1.2, 1:1.4, 1:1.5).

[0073] Specifically, the catalyst in step (2) is a zero-valent palladium complex, such as tetra(triphenylphosphine)palladium.

[0074] Specifically, the molar ratio of compound 3 to catalyst in step (2) is 1:0.01-0.1 (e.g., 1:0.02, 1:0.04, 1:0.05, 1:0.06, 1:0.08).

[0075] Specifically, the alkali mentioned in step (2) is selected from sodium carbonate, potassium carbonate, lithium carbonate and cesium carbonate, such as potassium carbonate.

[0076] Specifically, the molar ratio of compound 3 to the base in step (2) is 1:1.5-3 (e.g., 1:1.8, 1:2, 1:2.2, 1:2.5).

[0077] Specifically, the solvent in step (2) can be a mixture of 1,4-dioxane and water (e.g., 4:1, v / v).

[0078] Specifically, the reaction temperature in step (2) is 60-100℃ (e.g., 60, 70, 80, 85, 90, 95, 100℃), for example, 80-95℃.

[0079] Specifically, the reaction time in step (2) is 1-12 hours (e.g., 2, 3, 4, 5, 6, 8, 10, 12 hours).

[0080] Specifically, the carbonylating agent mentioned in step (3) can be hydrobromic acid, acyl chloride, carbonyl cuprous salt, etc., especially hydrobromic acid.

[0081] Specifically, the reaction temperature in step (3) is 80-120℃ (e.g., 90, 95, 100, 105, 110, 120℃), for example, 90-110℃.

[0082] Specifically, the reaction time in step (3) is 0.5-6 hours (e.g., 0.5, 1, 1.5, 2, 3, 4, 6 hours).

[0083] Furthermore, the preparation method may also include the preparation steps of compounds 1, 2, 4, and 4'.

[0084] Specifically, each step of the preparation method may further include a separation and purification step.

[0085] In a third aspect of the invention, a pharmaceutical composition is provided comprising the compound described in the first aspect or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate thereof, and one or more pharmaceutically acceptable excipients.

[0086] Specifically, in the pharmaceutical composition, the compound described in the first aspect or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate may be used alone or in combination with other types of active ingredients.

[0087] Specifically, the pharmaceutically acceptable excipients may be selected from one or more of the following: fillers, binders, lubricants, disintegrants, antioxidants, buffers, antibacterial agents, suspending agents, solubilizers, thickeners, stabilizers, and preservatives.

[0088] Specifically, the pharmaceutical composition can be administered via any suitable route of administration, such as gastrointestinal administration (e.g., oral, sublingual, rectal administration) or non-gastrointestinal administration (e.g., intravenous, intramuscular, intranasal, intraocular, intracerebral, intravaginal, intraperitoneal, transdermal, subcutaneous, intradermal, respiratory tract administration, etc.).

[0089] In some embodiments of the present invention, the pharmaceutical composition is an oral formulation, including, but not limited to, tablets (including sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets, oral tablets, etc.), pills, powders, granules, capsules (including soft capsules, microcapsules), lozenges, syrups, liquids, emulsions, suspensions, and controlled-release formulations (e.g., instantaneous-release formulations, sustained-release formulations, sustained-release microcapsules).

[0090] In some embodiments of the present invention, the pharmaceutical composition is an injectable preparation (e.g., subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection).

[0091] In other embodiments of the invention, the pharmaceutical composition is an intravenous infusion, a transdermal absorption formulation, a lotion, a suppository (e.g., a rectal suppository, a vaginal suppository), a nasal preparation, a pulmonary preparation (inhaler), an eye drop, etc.

[0092] Specifically, the pharmaceutical composition is preferably in unit dosage form. In this form, the formulation is further divided into unit doses containing an appropriate amount of the active ingredient. The unit dosage form can be a capsule, tablet, or any dosage form; alternatively, the unit dosage form can also be a packaged formulation, such as tablets, capsules, and powders packaged in vials or ampoules.

[0093] Specifically, the amount of the active ingredient in the unit dose formulation may be varied or adjusted from 0.1 mg to 1000 mg (e.g., 0.1, 1, 5, 10, 20, 40, 50, 100, 200, 400, 500, 1000 mg), depending on the specific application and potency of the active ingredient. If desired, the composition may also contain other suitable therapeutic agents.

[0094] In a fourth aspect of the invention, the use of the compound of the first aspect or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate thereof, or the pharmaceutical composition of the third aspect, in the preparation of a medicament for the prevention and / or treatment of PKMYT1-related diseases is provided.

[0095] Specifically, the disease is one in which the prevention and / or treatment of PKMYT1 is beneficial, particularly diseases involving CCNE1 gene amplification or CCNE1 overexpression.

[0096] Specifically, the diseases mentioned include tumors, inflammatory diseases, autoimmune diseases, infectious diseases, precancerous syndromes, etc.

[0097] In some embodiments of the present invention, the disease is a tumor, particularly a malignant tumor, including but not limited to solid tumors and hematologic malignancies, such as: acute myeloid leukemia, adolescent cancer, childhood adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendiceal cancer, astrocytoma, atypical teratoma, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem cell glioma, brain tumor, breast cancer, bronchial tumor, Burkitt lymphoma, carcinoid tumor, atypical teratoma, embryonal tumor, germ cell tumor, primary lymphoma, cervical cancer, childhood cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia. Blood disorders (CLL), chronic myeloid leukemia (CML), chronic myelodysplastic disorder, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), embryonal tumor, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal glioma, Ewing sarcoma, extracranial ectodermal cell tumor, gonadal ectodermal cell tumor, ocular cancer, osteofibrous histiocytoma, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastoma, piloblastic leukemia, head and neck cancer, heart cancer, liver cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor. Pancreatic neuroendocrine tumors, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic and occult primary squamous neck cancer, midline carcinoma, oral cancer, multiple endocrine tumor syndrome, multiple myeloma / plasma cell tumor, mycosis fungoides, myelodysplastic syndrome, spinal dysplasia / myeloproliferative neoplasm, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma and osteosarcoma of bone, nasal cavity and paranasal sinus cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer. Cancer, pancreatic cancer, papilloma, paraganglioma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pleural pulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, stomach / gastric cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, laryngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastoma, rare childhood cancers, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or virus-induced cancer.

[0098] In some embodiments of the present invention, the disease is a solid tumor, such as: colorectal cancer, gastric cancer, ovarian cancer, breast cancer, endometrial cancer, kidney cancer, prostate cancer, bladder cancer, liver cancer, lung cancer (such as non-small cell lung cancer), esophageal cancer, pancreatic cancer, cervical cancer, glioblastoma, and neuroblastoma.

[0099] In a fifth aspect of the invention, a method for inhibiting the growth of CCNE1 gene-amplified cells is provided, comprising the step of contacting the cells with the compound of the first aspect or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate thereof, or the pharmaceutical composition of the third aspect.

[0100] Specifically, the method is performed in vivo or in vitro.

[0101] In a sixth aspect of the invention, a method for preventing and / or treating PKMYT1-related diseases is provided, comprising administering to a subject in need the compound of the first aspect or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or, a pharmaceutical composition of the third aspect, the compound of the first aspect or thereof.

[0102] Specifically, the disease is as described in the fourth aspect of the present invention.

[0103] Specifically, the subjects are mammals, such as humans.

[0104] Specifically, the administration route can be any suitable route, such as gastrointestinal administration (e.g., oral, sublingual, rectal) or non-gastrointestinal administration (e.g., intravenous, intramuscular, intranasal, intraocular, intracerebral, intravaginal, intraperitoneal, transdermal, subcutaneous, intradermal, respiratory tract, etc.). In some embodiments of the present invention, the administration route is oral. In other embodiments of the present invention, the administration route is injection (e.g., subcutaneous injection, intravenous injection, intramuscular injection).

[0105] This invention provides a novel PKMYT1 kinase inhibitor with superior inhibitory activity. It can be used to inhibit the overactivation of CDK1 in CCNE1-amplified cells by inhibiting PKMYT1 kinase, thereby preventing the overactivation of CDK1 and triggering mitosis before DNA replication is completed. This leads to chromosome fragmentation, catastrophic DNA damage, and cell death. Consequently, it can be used for the prevention and / or treatment of related diseases such as cancer (especially endometrial cancer, ovarian cancer, non-small cell lung cancer, esophageal and gastric cancer). It can fill the gap in targeted drugs for cancers with specific CCNE1 amplification subtypes and has very good application prospects and value in the pharmaceutical field. Attached Figure Description

[0106] Figure 1 shows the inhibition curve of compound 15 on PKMYT1 enzyme activity.

[0107] Figure 2 shows the inhibition curve of compound 15 on the growth of human breast ductal carcinoma cells HCC1569. Detailed Implementation

[0108] Unless otherwise defined, all scientific and technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art.

[0109] In this invention, the term "aliphatic group" refers to a straight-chain or branched hydrocarbon chain that is fully saturated or contains one or more unsaturated units, or a cyclic hydrocarbon group (also referred to herein as an "aliphatic ring") that is fully saturated or contains one or more unsaturated units, connected to the rest of the molecule by a single bond. Suitable aliphatic groups include, but are not limited to, straight-chain or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl, and mixtures thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, (cycloalkyl)alkenyl, etc. Typical aliphatic groups contain 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 1 to 6 carbon atoms.

[0110] The term "carbon ring" is composed entirely of carbon atoms and can be divided into aliphatic rings and aromatic rings.

[0111] The term "alkyl" refers to a straight-chain or branched hydrocarbon radical that does not contain unsaturated bonds and is connected to the rest of the molecule by a single bond. Typical alkyl groups contain 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, isohexyl, etc. If the alkyl group is substituted with a cycloalkyl group, it is referred to as "cycloalkylalkyl," such as cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, etc. If the alkyl group is substituted with an aryl group, it is referred to as "aralkylalkyl," such as benzyl, diphenylmethyl, or phenethyl. If the alkyl group is substituted with a heterocyclic group, it is referred to as "heterocyclicalkyl." In this invention, CO alkyl refers to H, i.e., C 0-10 Alkyl (or C0-C) 10 Alkyl groups include H and C. 1-10 Alkyl (or C1-C) 10 alkyl).

[0112] The term "alkylene" refers to a hydrocarbon group (divalent alkyl) formed by the loss of two hydrogen atoms from an alkane molecule. It can be straight-chain or branched and is connected to the rest of the molecule by a single bond. Typical alkylene groups described herein have 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 1 to 6 carbon atoms, such as methylene (-CH2-), ethylene, propylene, butylene, etc. In this invention, CO alkylene refers to a single bond, i.e., C... 0-10 Alkylene (or C0-C) 10 Alkylenes include single bonds and C bonds. 1-10 Alkylene (or C1-C) 10(alkylene).

[0113] The term "alkoxy" refers to a substituent formed when a hydrogen atom in a hydroxyl group is replaced by an alkyl group. Typical alkoxy groups contain 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) carbon atoms, such as methoxy, ethoxy, propoxy, butoxy, etc.

[0114] The term "cycloalkyl" refers to a saturated or partially saturated (especially saturated) monocyclic or polycyclic group that may contain 1 to 4 monocyclic and / or fused rings and 3 to 18 carbon atoms, preferably 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or adamantyl.

[0115] The term "aryl" refers to a monocyclic or polycyclic free radical, including polycyclic free radicals containing a monoaryl group and / or a fused aryl group, such as those containing 1-3 monocyclic or fused rings and 6-18 (e.g., 6, 8, 10, 12, 14, 16, 18) carbon ring atoms. Typical aryl groups are those containing 6-12 carbon ring atoms, such as phenyl, naphthyl, biphenyl, and indenyl. "Arylidene" refers to a divalent group derived from aromatic hydrocarbons by removing two hydrogen atoms.

[0116] The term "heterocyclic group" includes heteroaromatic and heterocyclic groups containing 1 to 3 monocyclic and / or fused rings and 3 to 18 ring atoms. Preferred heteroaromatic and heterocyclic groups contain 5 to 10 ring atoms. Suitable heteroaryl groups in the compounds of the present invention contain 1, 2, or 3 heteroatoms selected from N, O, or S atoms. Examples of heteroaryl groups, such as, but not limited to, coumarins, including 8-coumarins; quinolinyl groups, including 8-quinolinyl, isoquinolinyl, pyridyl, pyrazinyl, pyrazolyl, pyrimidinyl, furanyl, pyrroloyl, thiopheneyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, imidazoleyl, indoleyl, isoindoleyl, indazoleyl, inazinyl, phthalazinyl, pteridinyl, purineyl, oxadiazolyl, thiadiazolyl, furazolidyl, pyridazinyl, triazinyl, cenolinyl, benzimidazolyl, benzofuranyl, benzofuranyl, benzothiopheneyl, benzothiazolyl, benzoxazolyl, quinazolinyl, naphridinyl, and furanopyridinyl, etc. Suitable heterocyclic groups in the compounds of the present invention contain one, two, or three heteroatoms selected from N, O, or S atoms. Examples of heterocyclic groups, such as, but not limited to, pyrrolidinyl, tetrahydrofuranyl, dihydrofuran, tetrahydrothiophenyl, tetrahydrothiophenyl, piperidinyl, morpholinyl, thiomorpholinyl, oxothiocyclohexyl, piperazine, aziridine, oxocyclobutyl, thiocyclobutyl, high-piperidinyl, oxocyclopropane, thiocyclopropane, acrylonitrile, oxoaziridine, diacylonitrile, triacylonitrile, 1,2,3,6-tetracyclyl Hydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, dihydroindolyl, 2H-pyranyl, 4H-pyranyl, dioxacyclohexyl, 1,3-dioxapentyl, pyrazolinyl, dithiaalkyl, dithiopentyl, dihydropyranyl, dihydrothiophenyl, pyrazolinyl, imidazolinyl, imidazolinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolyl, and quinazinyl, etc. In this invention, for optionally substituted heterocyclic groups, the substituted position can be any suitable carbon atom or heteroatom, for example, for The substitution position of R can be any suitable carbon or nitrogen atom, and it can be, for example...

[0117] The above-mentioned groups can be replaced by one or more suitable groups at one or more available positions, such as: OR', =O, SR', SOR', SO2R', OSO2R', OSO3R', NO2, NHR', N(R')2, =N-R', N(R')COR', N(COR')2, N(R')SO2R', N(R')C(=NR')N(R')R', N3, CN, halogen, COR', COOR', OCOR', OCOOR', OCONHR', OCON(R')2, CONHR', CON(R')2, CON(R')OR', CON(R')SO2R', PO(OR')2, PO(OR')R', PO(OR')(N(R')R'), Cl-C 12 Alkyl, C3-C 10 cycloalkyl, C2-C 12 alkenyl, C2-C 12 Alkynyl, aryl, and heterocyclic groups, wherein each R' group is independently selected from: hydrogen, OH, NO2, NH2, SH, CN, halogen, COH, COalkyl, COOH, C1-C 12 Alkyl, C3-C 10 cycloalkyl, C2-C 12 alkenyl, C2-C 12 Alkynyl, aryl, and heterocyclic groups. These groups are themselves substituted, and the substituents can be selected from the aforementioned list.

[0118] "Halogen" refers to bromine, chlorine, iodine, or fluorine. Haloalkyl refers to a group in which the hydrogen atom on the alkyl group is replaced by a halogen atom (F, Cl, Br, I), such as -CH2Rh, -CHRh2, -CRh3, where Rh is F, Cl, Br, or I; such as -CF3.

[0119] The term "pharmaceutically acceptable salt" refers to an acidic or basic salt that is theoretically non-toxic, non-irritating, and non-allergenic, and that can achieve or provide clinically acceptable pharmacokinetic, absorption, distribution, and metabolic properties of a drug molecule to achieve its intended purpose. The salts described in this invention include pharmaceutically acceptable acidic or basic salts of compounds with acidic, basic, or amphoteric groups. A list of suitable salts can be found in SM Birge, et al., J. Pharm. Sci., 66, 1-19 (1977).

[0120] The pharmaceutically acceptable salts described in this invention include acid addition salts and base addition salts.

[0121] The acid addition salts include, but are not limited to, salts from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, and phosphonic acid, as well as salts from organic acids such as aliphatic monocarboxylic acids and dicarboxylic acids, phenyl-substituted alkanic acids, hydroxyalkanic acids, alkanedioic acids, aromatic acids, and aliphatic and aromatic sulfonic acids. Therefore, these salts include, but are not limited to, sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, hydrochlorides, hydrobromates, iodates, acetates, propionates, octanoates, isobutyrates, oxalates, malonates, succinates, octanoates, sebacic acid salts, fumarates, maleates, amygdalinates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, maleates, tartrates, and methanesulfonates, as well as salts of amino acids such as arginine salts, gluconates, and galacturonic acids. Acid addition salts can be prepared by contacting a sufficient amount of the desired acid in a conventional manner to form a salt. The free base can be regenerated by contacting the salt with a base, and the free base can be separated in a conventional manner.

[0122] The base addition salts described in this invention refer to salts formed with metals or amines, such as hydroxides of alkali metals and alkaline earth metals, or with organic amines. Examples of metals used as cations include, but are not limited to, sodium, potassium, magnesium, and calcium. Suitable amines include, but are not limited to, N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine (ethane-1,2-diamine), N-methylglucosamine, and procaine. Base addition salts can be prepared by contacting a sufficient amount of the desired base in a conventional manner to form a salt. The free acid form can be regenerated by contacting the salt form with an acid, and the free acid can be separated in a conventional manner.

[0123] The term "solvent" should be understood to refer to any form of the compounds of the present invention, wherein the compounds are linked to another molecule (usually a polar solvent) by a non-covalent bond, particularly including hydrates and alcohols, such as methanols. Hydrates are preferred solvates.

[0124] The term "prodrug" is used in its broad sense and encompasses derivatives that can be converted into the compounds of this invention in vivo. Examples of prodrugs include, but are not limited to, derivatives and metabolites of compounds, including biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable acylureas, and biohydrolyzable phosphate ester analogs. Preferably, prodrugs having a carboxyl functional group are lower alkyl esters of carboxylic acids. The carboxylic acid esters are readily obtained by esterification of any carboxylic acid moiety present in the molecule. Prodrugs can generally be prepared by known methods, such as those described in Burger's "Medicinal Chemistry and Drug Discovery, 6th Edition" (Donald J. Abraham ed., 2001, Wiley) and "Design and Applications of Prodrugs" (H. Bundgaard ed., 1985, Harwood Academic Publishers).

[0125] The term "not present" indicates that the linking group is a linking bond.

[0126] Any compound referred to herein is intended to represent such a particular compound and certain variations or forms thereof. In particular, the compounds referred to herein may have an asymmetric center and therefore exist in different enantiomers or diastereomers. Thus, any given compound referred to herein represents any racemic compound, one or more enantiomers, one or more diastereomers, or mixtures thereof. Similarly, stereoisomers or geometric isomers of the double bonds may also exist, thus in some cases the molecule may exist as (E)-isomers or (Z)-isomers (trans and cis isomers). If the molecule contains multiple double bonds, then each double bond will have its own stereoisomerism, which may be the same as or different from the stereoisomerism of the other double bonds of the molecule. Furthermore, the compounds referred to herein may exist as ator isomers. All stereoisomers of the compounds referred to herein, including enantiomers, diastereomers, geometric isomers, and ator isomers, and mixtures thereof, are within the scope of this invention.

[0127] The term "tumor" refers to an abnormal mass of tissue that grows beyond and out of harmony with the growth of normal tissue. "Cancer" or "malignant tumor" refers to a disease characterized by uncontrolled growth and spread of malignant cells and tissue infiltration, and which is pathologically identified as a malignant tumor according to the "Classification of Diseases and Causes of Death" published by the Ministry of Health of China.

[0128] The term "autoimmune disease" refers to diseases caused by damage to the body's own tissues due to an immune response to its own antigens, such as, but not limited to: systemic lupus erythematosus, ulcerative colitis, Crohn's disease, multiple sclerosis, psoriasis, rheumatoid arthritis, etc.

[0129] The term "inflammation" refers to the body's defensive response to stimuli, manifested as redness, swelling, heat, pain, and functional impairment. It can be infectious inflammation caused by infection, or non-infectious inflammation not caused by infection, such as inflammation caused by immune responses (e.g., various types of hypersensitivity reactions, inflammation caused by some autoimmune diseases). The term "inflammatory disease" refers to a disease characterized by inflammation.

[0130] The term "infectious disease" mainly refers to diseases caused by pathogen infection, including symptoms of bodily damage and infection response caused by pathogen invasion.

[0131] All publications, patents, and published patent specifications cited in this article are incorporated herein in their entirety through citation.

[0132] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0133] Compound Synthesis Examples

[0134] Compounds 1 and 3-13 were synthesized according to the following route:

[0135] Compounds 15-31 were synthesized according to the following route:

[0136] Example 1: Synthesis of Compound 1

[0137] 1.1 Synthesis of Intermediate A-1

[0138] 3-Amino-2-methoxy-6-bromopyridine (CAS No.: 89466-18-2) (1.0 eq) was dissolved in N,N-dimethylformamide, followed by the addition of N-chlorosuccinimide (NCS) (1.2 eq), and the reaction was carried out at room temperature for 2 hours. After the reaction was completed as monitored by LC-MS, the reaction solution was diluted with ethyl acetate, washed once with saturated sodium bicarbonate, twice with water, and once with saturated brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate A-1.

[0139] 1.2 Synthesis of Intermediate B-1

[0140] Intermediate 4-bromo-7-fluoro-1H-indazole (CAS No.: 1186334-63-3) (1.0 eq), pinacol diboronate (CAS No.: 73183-34-3) (1.2 eq), Pd(PPh3)Cl2 (0.05 eq), and potassium acetate (KAcO) (2.5 eq) were added to dimethyl sulfoxide. The mixture was purged with nitrogen three times, and then heated to 120 °C for 16 hours. After the reaction was completed by LC-MS monitoring, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate B-1.

[0141] 1.3 Synthesis of intermediate MYI-001-1

[0142] Intermediate A-1 (1.0 eq), (4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzene (R-1, 1.1 eq), Pd(PPh3)4 (0.05 eq), and potassium carbonate (K2CO3) (2.0 eq) were added to 1,4-dioxane and water (4:1, v / v). The mixture was purged with nitrogen three times, and then heated to 90 °C for 4 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-001-1.

[0143] 1.4 Synthesis of intermediate MYI-001-2

[0144] Intermediate MYI-001-1 (1.0 eq), intermediate B-1 (1.2 eq), Pd(dtbpf)Cl2 (0.05 eq), and potassium carbonate (K2CO3) (2.0 eq) were added to 1,4-dioxane and water (4:1). The mixture was purged with nitrogen three times, and then the temperature was raised to 90 °C for 4 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-001-2.

[0145] 1.5 Synthesis of Compound 1

[0146] Intermediate MYI-001-2 (1.0 eq) was added to a 48% aqueous hydrobromic acid solution, and the mixture was then heated to 100 °C and reacted for 1 hour. After the reaction was completed by LC-MS monitoring, the system was adjusted to alkalinity with a saturated sodium bicarbonate solution, extracted with ethyl acetate, washed once with brine, and the resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Then, it was subjected to column chromatography to obtain compound 1.

[0147] Example 2: Synthesis of Compound 3

[0148] Following the synthesis method of Example 1, R-1 was replaced with 4-pyridineboronic acid pinacol ester (CAS No.: 181219-01-2) to prepare compound 3.

[0149] Example 3: Synthesis of Compound 4

[0150] Following the synthesis method of Example 1, R-1 was replaced with 3-pyridineboronic acid pinacol ester (CAS No.: 329214-79-1) to prepare compound 4.

[0151] Example 4: Synthesis of Compound 5

[0152] Following the synthesis method of Example 1, R-1 was replaced with 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (CAS No.: 287944-16-5) to prepare compound 5.

[0153] Example 5: Synthesis of Compound 6

[0154] Following the synthesis method of Example 1, R-1 was replaced with 1-methylpyrazole-3-boronic acid pinacol ester (CAS No.: 1020174-04-2) to prepare compound 6.

[0155] Example 6: Synthesis of Compound 7

[0156] Following the synthesis method of Example 1, R-1 was replaced with 1-acetyl-5,6-dihydro-2H-pyridine-4-boronic acid pinacol ester (CAS No.: 1227068-67-8), and B-1 was replaced with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1H-indazole (CAS No.: 885618-33-7) to prepare compound 7.

[0157] Example 7: Synthesis of Compound 8

[0158] Following the synthesis method of Example 1, R-1 was replaced with 1-methyl-1,2,3,6-tetrahydropyridine-4-boronic acid pinacol ester (CAS No.: 454482-11-2) to prepare compound 8.

[0159] Example 8: Synthesis of Compound 9

[0160] Following the synthesis method of Example 1, R-1 was replaced with 6-methoxypyridine-2-boronic acid pinacol ester (CAS No.: 1034297-69-2) to prepare compound 9.

[0161] Example 9: Synthesis of Compound 2

[0162] 1.1 Synthesis of intermediate MYI-002-1

[0163] At 0°C, sodium methoxide (3.0 eq) was added to a methanol solution of 4,6-dichloro-2-(propylthio)-5-aminopyrimidine (CAS No.: 145783-15-9) (1.0 eq), and the mixture was then heated to room temperature and reacted for 2 hours. After the reaction was completed as monitored by LC-MS, the mixture was extracted with ethyl acetate, washed once with brine, and the resulting organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-002-1.

[0164] 1.2 Synthesis of intermediate MYI-002-2

[0165] Intermediate MYI-002-1 (1.0 eq), intermediate B-1 (1.2 eq), Pd(dtbpf)Cl2 (0.05 eq), and potassium carbonate (K2CO3) (2.0 eq) were added to 1,4-dioxane and water (4:1). The mixture was purged with nitrogen three times, and then the temperature was raised to 80 °C for 4 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-002-2.

[0166] 1.3 Synthesis of intermediate MYI-002-3

[0167] Intermediate MYI-002-2 (1.0 eq), di-tert-butyl dicarbonate ((Boc)₂O) (3.3 eq), 4-dimethylaminopyridine (DMAP) (0.05 eq), and triethylamine (TEA) (4.0 eq) were added to THF. The mixture was purged with nitrogen three times, and then the temperature was raised to 40 °C for 4 hours. After the reaction was completed by LC-MS monitoring, the reaction solution was diluted with ethyl acetate, washed twice with water, and washed once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-002-3.

[0168] 1.4 Synthesis of intermediate MYI-002-4

[0169] Intermediate MYI-002-3 (1.0 eq), pinacol 3-pyridineborate (1.2 eq), cuprous thiophene-2-carboxylate (I)(CuTc)(2e.q.), and Pd(PPh3)4 (0.05 eq) were added to methanol, purged three times with nitrogen, and then the mixture was heated to 30 °C and reacted for 16 hours. After the reaction was completed by LC-MS monitoring, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-002-4.

[0170] 1.5 Synthesis of Compound 2

[0171] Intermediate MYI-002-4 (1.0 eq) was added to a 48% aqueous hydrobromic acid solution (10.0 eq), and the mixture was then heated to 100 °C and reacted for 1 hour. After the reaction was completed by LC-MS monitoring, the system was adjusted to alkalinity with a saturated sodium bicarbonate solution, extracted with ethyl acetate, washed once with brine, and the resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Then, it was subjected to column chromatography to obtain compound 2.

[0172] Example 10: Synthesis of Compound 10

[0173] 1.1 Synthesis of intermediate C-1

[0174] 3-Bromo-2,4-dimethylphenol (1.0 eq) and K₂CO₃ (2.5 eq) were added to acetone, and dimethyl sulfate (1.5 eq) was added dropwise three times under nitrogen purging. The mixture was then heated to 60 °C and reacted for 4 hours. After the reaction was monitored by LC-MS, the reaction solution was diluted with ethyl acetate, washed twice with water and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate C-1.

[0175] 1.2 Synthesis of intermediate C-2

[0176] Intermediate C-1 (1.0 eq) was added to a solution of tetrahydrofuran (THF), purged three times with nitrogen, and n-butyllithium (1.2 eq) was added dropwise at -70 °C. The reaction mixture was stirred at the same temperature for 30 minutes, followed by dropwise addition of trimethylboronic acid ester (B(OCH3)3) (1.5 eq). Stirring was continued at the same temperature for another 30 minutes, followed by slow warming to room temperature overnight. TLC showed the reaction was complete. The reaction mixture was quenched with saturated 1M HCl (aq.) and then diluted with water. The mixture was extracted with ethyl acetate. The combined organic layers were washed with brine and dried over anhydrous sodium sulfate to give C-2, a yellow oil.

[0177] 1.3 Synthesis of intermediates MYI-010-1 and MYI-010-2

[0178] Following the synthesis method of Example 1, R-1 was replaced with 4-pyridineboronic acid pinacol ester (CAS No.: 181219-01-2), and B-1 was replaced with intermediate C-2 to prepare intermediates MYI-010-1 and MYI-010-2.

[0179] 1.4 Synthesis of Compound 10

[0180] Intermediate MYI-010-2 (1.0 eq) was added to a solution of dichloromethane (DCM), purged three times with nitrogen, and boron tribromide (BBr3) (6.0 eq) was added dropwise at 0 °C. The mixture was then stirred at the same temperature for 30 minutes, followed by slow heating to room temperature overnight. LC-MS showed the reaction was complete. The reaction mixture was quenched with methanol, concentrated under reduced pressure, and then diluted with water. The mixture was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and then subjected to column chromatography to give compound 10.

[0181] Example 11: Synthesis of Compound 11

[0182] 1.1 Synthesis of intermediate MYI-011-1

[0183] 4-Bromo-2-fluoropyridine (CAS No.: 128071-98-7) (1.0 eq), tert-butyl 1,4-diazacycloheptan-1-carboxylate (2.0 eq), and K2CO3 (3.0 eq) were added to DMSO, purged with nitrogen three times, and then heated to 100℃ for 4 hours. After the reaction was completed by LC-MS monitoring, the reaction solution was diluted with ethyl acetate, washed twice with water, and washed once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-011-1.

[0184] 1.2 Synthesis of intermediate MYI-011-2

[0185] Intermediate MYI-011-1 (1.0 eq), pinacol diborate (1.2 eq), Pd(dppf)Cl2 (0.05 eq), and potassium acetate (KAcO) (2.5 eq) were added to dioxane. The mixture was purged with nitrogen three times, and then heated to 100 °C for 6 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-011-2.

[0186] 1.3 Synthesis of Compound 11

[0187] Following the synthesis method of Example 1, R-1 was replaced with MYI-011-2 to prepare compound 11.

[0188] Example 12: Synthesis of Compound 12

[0189] 1.1 Synthesis of intermediate MYI-012-1

[0190] 4-Bromo-2-fluoropyridine (CAS No.: 128071-98-7) (1.0 eq), (R)-3-tert-butoxycarbonylaminopyrrolidine (2.0 eq), and K2CO3 (3.0 eq) were added to DMSO, purged with nitrogen three times, and then heated to 100℃ for 4 hours. After the reaction was completed by LC-MS monitoring, the reaction solution was diluted with ethyl acetate, washed twice with water and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-012-1.

[0191] 1.2 Synthesis of intermediate MYI-012-2

[0192] Intermediate MYI-012-1 (1.0 eq), pinacol diborate (1.2 eq), Pd(dppf)Cl2 (0.05 eq), and potassium acetate (KAcO) (2.5 eq) were added to dioxane. The mixture was purged with nitrogen three times, and then heated to 100 °C for 6 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-012-2.

[0193] 1.3 Synthesis of Compound 12

[0194] Following the synthesis method of Example 1, R-1 was replaced with MYI-012-2 to prepare compound 12.

[0195] Example 13: Synthesis of Compound 13

[0196] 1.1 Synthesis of intermediate MYI-013-1

[0197] 4-Bromo-2,6-fluoropyridine (CAS No.: 903513-58-6) (1.0 eq), tert-butyl 1,4-diazacycloheptan-1-carboxylate (2.0 eq), and K2CO3 (3.0 eq) were added to DMSO, purged with nitrogen three times, and then heated to 100℃ for 4 hours. After the reaction was completed by LC-MS monitoring, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-013-1.

[0198] 1.2 Synthesis of intermediate MYI-013-2

[0199] Intermediate MYI-013-1 (1.0 eq), pinacol diboronate (1.2 eq), Pd(dppf)Cl2 (0.05 eq), and potassium acetate (KAcO) (2.5 eq) were added to dioxane. The mixture was purged with nitrogen three times, and then heated to 100 °C for 6 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-013-2.

[0200] 1.3 Synthesis of Compound 13

[0201] Following the synthesis method of Example 1, R-1 was replaced with intermediate MYI-013-2, and B-1 was replaced with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1H-indazole (CAS No.: 885618-33-7) to prepare compound 13.

[0202] Example 14: Synthesis of Compound 14

[0203] 1.1 Synthesis of intermediate MYI-014-1

[0204] Intermediate A-1 (1.0 eq), 7H-pyrrolo[2,3-D]pyrimidin-2-amine (CAS No.: 93366-88-2) (2.0 eq), copper iodide (3.0 eq), trans-1,2-cyclohexanediamine (0.4 eq), and K3PO4 (2.0 eq) were added to dioxane. The mixture was purged with nitrogen three times, and then heated to 100 °C for 16 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-014-1.

[0205] 1.2 Synthesis of Compound 14

[0206] Compound 14 was prepared by referring to the synthesis method of Example 1.

[0207] Example 15: Synthesis of Compound 15

[0208] 1.1 Synthesis of Intermediate A-2

[0209] Intermediate A-1 (1.0 eq), pinacol diborate (1.2 eq), Pd(dppf)Cl2 (0.05 eq), and potassium acetate (KAcO) (2.5 eq) were added to DMSO. The mixture was purged with nitrogen three times, and then the temperature was raised to 100 °C for 6 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate A-2.

[0210] 1.2 Synthesis of intermediate MYI-015-1

[0211] Intermediate A-2 (1.0 eq), 2-bromothiazole (CAS No.: 3034-53-5) (R-1, 1.1 eq), Pd(PPh3)4 (0.05 eq), and potassium carbonate (K2CO3) (2.0 eq) were added to 1,4-dioxane and water (4:1, v / v). The mixture was purged with nitrogen three times, and then heated to 90 °C for 4 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-015-1.

[0212] 1.3 Synthesis of intermediate MYI-015-2

[0213] Intermediate MYI-015-1 (1.0 eq), intermediate B-1 (1.2 eq), Pd(dtbpf)Cl2 (0.05 eq), and potassium carbonate (K2CO3) (2.0 eq) were added to 1,4-dioxane and water (4:1, v / v), purged with nitrogen three times, and then heated to 90 °C for 4 hours. After the reaction was completed by LC-MS monitoring, the reaction solution was diluted with ethyl acetate, washed twice with water and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-015-2.

[0214] 1.4 Synthesis of Compound 15

[0215] Intermediate MYI-015-2 (1.0 eq) was added to a 48% aqueous hydrobromic acid solution, and the mixture was then heated to 100 °C and reacted for 1 hour. After the reaction was completed by LC-MS monitoring, the system was adjusted to alkalinity with a saturated sodium bicarbonate solution, extracted with ethyl acetate, washed once with brine, and the resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Then, it was subjected to column chromatography to obtain compound 15.

[0216] Example 16: Synthesis of Compound 16

[0217] Following the synthetic method of Example 15, R-1 was replaced with 2-bromo-4-cyanothiazole (CAS No.: 181219-01-2) to prepare a cyano compound. Finally, under nitrogen protection, the cyano compound was dissolved in 1,4-dioxane, and dilute hydrochloric acid was added. The mixture was stirred at 0-25°C and gradually heated to 40°C until the starting material disappeared. After the reaction was complete, the mixture was cooled, the pH was adjusted to neutral, and the organic solvent was removed under reduced pressure. The residue was extracted with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to obtain the corresponding formamide product, which is compound 16.

[0218] Example 17: Synthesis of Compound 17

[0219] Following the synthesis method of Example 15, R-1 was replaced with 2-bromo-4-cyanothiazole (CAS No.: 181219-01-2) to obtain a cyano compound. Concentrated sulfuric acid was added and the temperature was raised to 60°C, causing the cyano group to hydrolyze sequentially and eventually convert to formic acid. After the reaction was completed, the mixture was cooled, insoluble matter was removed by filtration, the pH was adjusted and then acidified to convert to formate. After crystallization and purification, the formic acid product compound 17 was obtained.

[0220] Example 18: Synthesis of Compound 18

[0221] Following the synthesis method of Example 15, R-1 was replaced with 4-bromopyridazine (CAS No.: 115514-66-4) to prepare compound 18.

[0222] Example 19: Synthesis of Compound 19

[0223] Following the synthesis method of Example 15, R-1 was replaced with 2-amino-6-bromopyridine (CAS No.: 19798-81-3), and B-1 was replaced with 1H-indazole-4-boronic acid pinacol ester (CAS No.: 885618-33-7) to prepare compound 19.

[0224] Example 20: Synthesis of Compound 20

[0225] Following the synthesis method of Example 15, R-1 was replaced with 2-bromobenzothiazole (CAS: 2516-40-7) to prepare compound 20.

[0226] Example 21: Synthesis of Compound 21

[0227] Following the synthesis method of Example 15, R-1 was replaced with tert-butyl 2-bromo-6,7-dihydrothiazo[4,5-C]pyridine-5(4H)-carboxylate (CAS No.: 1253654-37-3), and B-1 was replaced with intermediate B-2 to prepare compound 21.

[0228] Example 22: Synthesis of Compound 22

[0229] Following the synthesis method of Example 15, R-1 was replaced with 2-bromo-4-phenylthiazole (CAS: 57516-16-2) to prepare compound 22.

[0230] Example 23: Synthesis of Compound 23

[0231] Following the synthesis method of Example 15, R-1 was replaced with 2-bromo-4-trifluoromethylthiazole (CAS: 41731-39-9) to prepare compound 23.

[0232] Example 24: Synthesis of Compound 24

[0233] Following the synthesis method of Example 15, R-1 was replaced with 5-bromoisothiazol (CAS: 54390-97-5) to prepare compound 24.

[0234] Example 25: Synthesis of Compound 25

[0235] Following the synthesis method of Example 15, R-1 was replaced with 5-bromo-1-methyl-1H-imidazole (CAS: 1003-21-0) to prepare compound 25.

[0236] Example 26: Synthesis of Compound 26

[0237] 1.1 Synthesis of intermediate MYI-026-1

[0238] 2-Amino-6-bromopyridine (CAS No.: 19798-81-3) (2.0 eq), thiazolyl-2-carboxylic acid (1.0 eq), HATU (1.5 eq), and DIEA (2.0 eq) were added to DMF, purged three times with nitrogen, and then reacted at room temperature for 4 hours. After the reaction was monitored by LC-MS, the reaction solution was diluted with ethyl acetate, washed twice with water and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-026-1.

[0239] 1.2 Synthesis of Compound 26

[0240] Compound 26 was prepared by replacing R-1 with MYI-026-1 and B-1 with 1H-indazole-4-boronic acid pinacol ester (CAS No.: 885618-33-7) according to the synthesis method of Example 15.

[0241] Example 27: Synthesis of Compound 27

[0242] 1.1 Synthesis of intermediate MYI-027-1

[0243] 2-Bromo-4-thiazolic acid (CAS No.: 5198-88-9) (1.0 eq), N-methylpiperazine (2.0 eq), HATU (1.5 eq), and DIEA (2.0 eq) were added to DMF, and the mixture was reacted at room temperature for 4 hours. After the reaction was completed as monitored by LC-MS, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-027-1.

[0244] 1.2 Synthesis of Compound 27

[0245] Following the synthesis method of Example 15, R-1 was replaced with MYI-027-1 and B-1 was replaced with intermediate B-2 to prepare compound 27.

[0246] Example 28: Synthesis of Compound 28

[0247] 1.1 Synthesis of intermediate MYI-028-1

[0248] 2,6-Dibromopyridine (CAS No.: 626-05-1) (1.0 eq), N-methylpiperazine (2.0 eq), and K3PO4 (3.0 eq) were added to dioxane, purged three times with nitrogen, and then the mixture was heated to 80 °C and reacted for 4 hours. After the reaction was completed by LC-MS monitoring, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-028-1.

[0249] 1.2 Synthesis of Compound 28

[0250] Following the synthesis method of Example 15, R-1 was replaced with MYI-028-1 and B-1 was replaced with 1H-indazole-4-boronic acid pinacol ester (CAS No.: 885618-33-7) to prepare compound 28.

[0251] Example 29: Synthesis of Compound 29

[0252] 1.1 Synthesis of intermediate MYI-029-1

[0253] Intermediate A-1 (1.0 eq), 1H-pyrazole-3-boronate pinacol ester (CAS No.: 844501-71-9) (1.2 eq), Pd(PPh3)4 (0.05 eq), and K2CO3 (2.5 eq) were added to dioxane, purged three times with nitrogen, and then heated to 80 °C for 6 hours. After the reaction was completed by LC-MS monitoring, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-029-2.

[0254] 1.2 Synthesis of intermediate MYI-029-2

[0255] Intermediate MYI-029-1 (1.0 eq), 3-iodooxetane (CAS No.: 26272-85-5) (2.0 eq), and Cs₂CO₃ (3.0 eq) were added to DMF, purged three times with nitrogen, and then heated to 60 °C for 4 hours. After the reaction was completed by LC-MS monitoring, the reaction solution was diluted with ethyl acetate, washed twice with water and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-029-2.

[0256] 1.3 Synthesis of intermediate MYI-029-3

[0257] Intermediate MYI-029-2 (1.0 eq), B-1 (1.5 eq), Pd(dtbpf)Cl2 (0.05 eq), and K2CO3 (2.5 eq) were added to 1,4-dioxane and water (4:1, v / v), purged with nitrogen three times, and then the mixture was heated to 80 °C and reacted for 6 hours. After the reaction was monitored by LC-MS, the reaction solution was diluted with ethyl acetate, washed twice with water and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-029-3.

[0258] 1.4 Synthesis of intermediate MYI-029-4

[0259] Intermediate MYI-029-3 (1.0 eq) was added to a 48% aqueous hydrobromic acid solution, and the mixture was then heated to 100 °C and reacted for 1 hour. After the reaction was completed by LC-MS monitoring, the system was adjusted to alkalinity with a saturated sodium bicarbonate solution, extracted with ethyl acetate, washed once with brine, and the resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude intermediate MYI-029-4.

[0260] 1.5 Synthesis of Compound 29

[0261] Intermediate MYI-029-4 (1.0 eq), sodium hydroxide (2.0 eq), and TEBA (0.1 eq) were added to H2O, and the mixture was then heated to 60 °C and reacted for 4 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed twice with water, and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain compound 29.

[0262] Example 30: Synthesis of Compound 30

[0263] 1.1 Synthesis of intermediate MYI-030-1

[0264] 2-Amino-6-bromopyridine (CAS No.: 30683-23-9) (1.0 eq), N-methylpiperazine (2.0 eq), HATU (1.5 eq), and DIEA (2.0 eq) were added to DMF, purged three times with nitrogen, and then reacted at room temperature for 4 hours. After the reaction was monitored by LC-MS, the reaction solution was diluted with ethyl acetate, washed twice with water and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, followed by column chromatography to obtain intermediate MYI-030-1.

[0265] 1.2 Synthesis of Compound 30

[0266] Following the synthesis method of Example 15, R-1 was replaced with MYI-030-1 to prepare compound 30.

[0267] Example 31: Synthesis of Compound 31

[0268] 1.1 Synthesis of intermediate MYI-031-1

[0269] 4-Bromonicotinic acid (CAS No.: 15366-62-8) (1.0 eq), N-methylpiperazine (2.0 eq), HATU (1.5 eq), and DIEA (2.0 eq) were added to DMF, purged three times with nitrogen, and then reacted at room temperature for 4 hours. After the reaction was monitored by LC-MS, the reaction solution was diluted with ethyl acetate, washed twice with water and once with brine. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and then subjected to column chromatography to obtain intermediate MYI-031-1.

[0270] 1.2 Synthesis of Compound 31

[0271] Following the synthesis method of Example 15, R-1 was replaced with MYI-031-1 to prepare compound 31.

[0272] The structures and general analytical data of compounds 1-31 are shown in the table below.

[0273] Table 1. List of compounds and general analytical data

[0274] Bioactivity test examples

[0275] Example 1: Cell Experiment

[0276] 1. Experimental Methods

[0277] 1.1 Cell Seeding

[0278] Human breast ductal carcinoma cells HCC1569 (IM-H636) were cultured in RPMI 1640 medium (C3010-0500) containing a mixture of 15% fetal bovine serum (FBS) and 1% penicillin-streptomycin (C3421-0100) at 37°C and 5% CO2. The medium in a 10cm cell culture dish was aspirated, and the cells were rinsed twice with 1mL PBS solution. Then, 2mL of trypsin (C3530-0500) digestion solution was added, and the dish was incubated at 37°C for 3 minutes. Once the cell edges became rounded, 2mL of medium containing 15% FBS was added to stop the cell digestion process, and the cells were pipetted to form a cell suspension. Cell counts were performed, and the cell density was then diluted to 2×10⁶ cells / mL using complete culture medium. 4 cells / mL. Add 100 μL of cell suspension to each well of a white transparent 96-well plate (Coring, CLS3903) (add PBS solution to the edge of the well), and incubate the cell plate overnight in an incubator at 37°C and 5% CO2.

[0279] 1.2 Compound Preparation

[0280] The compound stock solution (100 mM DMSO stock solution) was diluted to 10 mM secondary stock solution using DMSO. 1 μL of the secondary stock solution was added to 1 mL of RPMI 1640 complete culture medium and shaken on a shaker to dissolve, thus preparing a maximum dosing concentration of 10 μM. 8 to 10 dosing concentrations were prepared by dilution at a 4-fold ratio and thoroughly mixed. The control group was prepared with 0.1% DMSO solvent.

[0281] 1.3 Drug treatment

[0282] After the cells adhered, the culture medium was carefully aspirated, and 100 μL of different concentrations of drug molecules prepared in RPMI 1640 complete culture medium was added to each well of a 96-well plate, with three replicates for each concentration. The cell plates were then incubated at 37°C in a 5% CO2 incubator for 7 days.

[0283] 1.4 Bioanalytical Methods

[0284] After 7 days of incubation, add 50 μL to each well. The assay reagent (Promega, G7572) was shaken to mix and incubated at room temperature for 10 min. The chemiluminescence value (RLU) was measured using a microplate reader. Cell viability at different drug concentrations was calculated using the following formula. The IC50 was obtained by fitting four parameters using GraphPad Prism. 50 Numerical value. Cell viability (%) = (As - Ab) / (Ac - Ab) * 100

[0285] As: Experimental RLU (containing cells, culture medium, and drug molecules);

[0286] Ac: Control group RLU (containing cells, culture medium and DMSO solvent);

[0287] Ab: Zeroing well RLU (containing culture medium only).

[0288] 2. Experimental Results

[0289] The experimental results are shown in Table 2.

[0290] Example 2: Enzyme Activity Experiment

[0291] 1. Experimental Methods

[0292] Determination of the inhibitory effect of the test substance on PKMYT1 ATPase activity:

[0293] The protein used was PKMYT1 (76-362aa) expressed in the *E. coli* system, with a 6xHis tag and Thrombin restriction site fused to its N-terminus. ADP-GLO enzyme activity assays were performed in 384-well white microplates (LABSELECT, 31432). The assay buffer consisted of 20 mM Tris, 3 mM MgCl2, 3 mM MnCl2, 50 mg / mL PEG20,000, 1 mM DTT, and pH 7.5.

[0294] Compound preparation: Dilute the stock solution of the compound (100mM DMSO stock solution) with DMSO to 50 times the highest concentration of the experimental compound, and then dilute it three times with DMSO in sequence to prepare 10 compound gradients.

[0295] Enzyme activation reaction system (44 μL pro + 1 μL cpds + 5 μL ATP): 44 μL of PKMYT1 protein solution (final concentration 62.5 nM) was dispensed into each well using a multichannel pipette, followed by 1 μL of the compound solution. Incubation was carried out at room temperature for 15 min. A control group containing protein and 1 μL DMSO was established, while a blank group containing only buffer was used. After incubation, 5 μL of ATP (final concentration 10 mM) was added to initiate the enzyme activation reaction, and incubation was carried out at room temperature for 60 min.

[0296] ADP-Glo TM Reaction system (5 μL enzyme activation reaction solution + 5 μL ADP-Glo TM Reagent + 10μL ADP-Glo TM Detection: After the enzyme activity reaction is complete, aliquot 5 μL of the enzyme activity reaction solution into a 384-well white microplate, setting up two replicates. Immediately add 5 μL of LADP-GloTM Reagent was added, and the reaction was allowed to proceed for 40 minutes. Then, 10 μL of Kinase Detection Reagent was added, and the reaction was allowed to proceed for another 30 minutes. RLU was detected using a microplate reader. The data were analyzed according to the following formula, and IC50 was obtained using a GraphPad Prism four-parameter fitting method. 50 Enzyme activity value. Inhibition rate (%) = 1 - (As-Ab) / (Ac-Ab) * 100

[0297] As: Experimental RLU (containing protein, ATP, and drug molecules);

[0298] Ac: Control group RLU (containing protein, ATP and DMSO solvent);

[0299] Ab: Blank group RLU (containing only ATP).

[0300] 2. Experimental Results

[0301] The experimental results are shown in Table 2.

[0302] Table 2 Biochemistry and Cellular Activity of Compounds Note: ++++: IC50 < 100 nM; +++: 100 nM ≤ IC50 < 500 nM; ++: 500 nM ≤ IC50 < 1 μM; +: 1 μM ≤ IC50 < 10 μM

[0303] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications or equivalent substitutions made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

[0304] The foregoing embodiments and methods described in this invention may vary based on the capabilities, experience, and preferences of those skilled in the art.

[0305] Listing the steps of the method in a certain order in this invention does not constitute any restriction on the order of the method steps.

Claims

1. A compound or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof, said compound having the following structure: in, X is selected from: N, CH; Ring A is a 3-15 membered carbon ring or a heterocyclic ring; R1, R2, R3, R4, R1', R2', R3', and R4' are independently selected from: H, D, halogen, cyano, nitro, and C1-C. 10 Alkyl, C3-C 10 cycloalkyl, C1-C 10 Haloalkyl, -O(C) 0-10 alkyl), -N(C) 0-10 Alkyl)(C 0-10 alkyl); R a It is one or more independent substituents on ring A, selected from: H, D, =O, halogen, cyano, nitro, C1-C. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl group, -L0-(C3-C 10 cycloalkyl), -L0-(C6-C 10 aryl), -L0-(4-10 membered heterocyclic group), -N(C 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl) (C3-C 10 cycloalkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -O (C3-C) 10 cycloalkyl), -S(C 0-10 Alkyl), -S(C3-C 10 cycloalkyl), -SO(C 0-10 alkyl), -SO2(C 0-10 Alkyl group), -SO2 (C3-C) 10 cycloalkyl), -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -SO2N(C 0-10 Alkyl) (C3-C 10 cycloalkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 Alkyl), -CON(C) 0-10 Alkyl) (C3-C 10 cycloalkyl), -CO(C 0-10 alkyl), -Si(C) 0-10 Alkyl)(C 0-10 Alkyl)(C 0-10 Alkyl); wherein, the C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl group, C3-C 10 cycloalkyl, C6-C 10 The hydrogen atom on the aryl or 4-10 membered heterocyclic group is optionally substituted by a group selected from the following: =O, halogen, cyano, nitro, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C3-C6 alkylene) 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl) (C3-C 10 cycloalkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -O (C3-C) 10 cycloalkyl), -S(C 0-10 Alkyl), -S(C3-C 10 cycloalkyl), -SO(C 0-10 alkyl), -SO2(C 0-10 Alkyl group), -SO2 (C3-C) 10 cycloalkyl), -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -SO2N(C 0-10 Alkyl) (C3-C 10 cycloalkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 Alkyl), -CON(C) 0-10 Alkyl) (C3-C 10 cycloalkyl), -CO(C 0-10 alkyl), -Si(C) 0-10 Alkyl)(C 0-10 Alkyl)(C 0-10 alkyl); L0 is selected from: single bond, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkyneylene, C3-C7 cycloalkylene, O, S, NH, CONH, NHCO, C(O); wherein the H on the C1-C6 alkylene and C3-C7 cycloalkylene groups is optionally substituted by a group selected from: halogen, cyano, hydroxyl, amino.

2. The compound according to claim 1, characterized in that, Ring A is a 3-8 member aliphatic ring, aromatic ring, 4-12 member heteroaromatic ring, or a 3-12 member saturated or partially unsaturated heterocycle; Preferably, Some are selected from the following structure: More preferably, Some have the following structure:

3. The compound according to claim 1 or 2, characterized in that, Each R a Independently selected from: H, =O, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, phenyl, 4-10 membered heterocyclic group, -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 alkyl), -N(C) 0-6 Alkyl)CO(C 0-6 Alkyl), -CON(C) 0-6 Alkyl)(C 0-6 alkyl), -CO(C) 0-6 alkyl), -COO(C 0-6 Alkyl), -OCO(C 1-6 Alkyl), -SO(C) 1-6 alkyl), -SO2(C 1-6 Alkyl); wherein the H on the C1-C6 alkyl, C3-C7 cycloalkyl, phenyl, or 4-10 membered heterocyclic group is optionally substituted by a group selected from the following groups: halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 alkyl), -N(C) 0-6 Alkyl)CO(C 0-6 alkyl), -CO(C) 0-6 Alkyl), -CON(C) 0-6 Alkyl)(C 0-6 alkyl), -COO(C 0-6 Alkyl), -OCO(C 1-6 Alkyl), -SO(C) 1-6 alkyl), -SO2(C 1-6 Alkyl); or, R a It has the following structure: R5 is selected from: C1-C6 alkyl, C3-C7 cycloalkyl, phenyl, 4-8 membered saturated heterocyclic groups; wherein the H on the alkyl, cycloalkyl, phenyl, or saturated heterocyclic group is optionally substituted by a group selected from: -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 Alkyl); or, R a It has the following structure: Wherein, L1 is selected from: C0-C6 alkylene, C3-C7 cycloalkylene, and R6 is selected from: H, halogen, cyano, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, phenyl, 4-10 membered heterocyclic group, -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 alkyl), -N(C) 0-6 Alkyl)CO(C 0-6 Alkyl), -CON(C) 0-6 Alkyl)(C 0-6 alkyl), -CO(C) 0-6 alkyl), -COO(C 0-6 Alkyl), -OCO(C 1-6 Alkyl), -SO(C) 1-6 alkyl), -SO2(C 1-6 Alkyl); wherein the H on the C1-C6 alkyl, C3-C7 cycloalkyl, phenyl, or 4-10 membered heterocyclic group is optionally substituted by a group selected from the following groups: halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 alkyl), -N(C) 0-6 Alkyl)CO(C 0-6 alkyl), -CO(C) 0-6 Alkyl), -CON(C) 0-6 Alkyl)(C 0-6 alkyl), -COO(C 0-6 Alkyl), -OCO(C 1-6 Alkyl), -SO(C) 1-6 alkyl), -SO2(C 1-6 Alkyl); or, R a It has the following structure: -L2-R7, where L2 is selected from: NH, CONH, NHCO, C(O); R7 is selected from: C3-C7 cycloalkyl, phenyl, 4-10 membered heterocyclic group, wherein the H on the cycloalkyl, phenyl, or 4-10 membered heterocyclic group is optionally substituted by a group selected from: halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, -N(C 0-6 Alkyl)(C 0-6 Alkyl), -O(C) 0-6 alkyl), -N(C) 0-6 Alkyl)CO(C 0-6 alkyl), -CO(C) 0-6 Alkyl), -CON(C) 0-6 Alkyl)(C 0-6 alkyl), -COO(C 0-6 Alkyl), -OCO(C 1-6 Alkyl), -SO(C) 1-6 alkyl), -SO2(C 1-6 alkyl); Preferably, each R a Independently selected from: H, =O, F, Cl, Br, I, CN, methyl, ethyl, isopropyl, -CF3, -CHF2, -CH2F, -OH、 4. The compound according to any one of claims 1-3, characterized in that, R1 is selected from: H, halogen, cyano, C1-C6 alkyl, C3-C7 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy; Preferably, R1 is selected from: H, halogen (such as F).

5. The compound according to any one of claims 1-4, characterized in that, R2 is selected from: H, halogens; and / or, R3 is selected from: H, halogens, C1-C6 alkyl groups; and / or, R4 is selected from: H, halogens, and C1-C6 alkyl groups.

6. The compound according to any one of claims 1-3, characterized in that, R1', R2', R3', and R4' are independently selected from: H, halogen, cyano, C1-C6 alkyl, C3-C7 cycloalkyl, C1-C6 haloalkyl, and C1-C6 alkoxy.

7. The compound according to claim 1, characterized in that, The compound is selected from the following structures:

8. A method for preparing the compound according to any one of claims 1-7, comprising the following steps: (1) Compound 1, Compound 2, catalyst, base and solvent are mixed and reacted to obtain Compound 3; (2) Compound 3, compound 4, catalyst, base and solvent are mixed and reacted to obtain compound 5; (3) Compound 5 was mixed with a carbonylating agent and reacted to obtain compound 6; or, (1) Compound 1, Compound 2, catalyst, base and solvent are mixed and reacted to obtain Compound 3; (2) Compound 3, compound 4', catalyst, base and solvent are mixed and reacted to obtain compound 5'; (3) Compound 5' was mixed with a carbonylating agent and reacted to obtain compound 6'; in, R P1 It is an alkyl group; R P2 It is a halogen or a thiol group; R P3 It is a halogen or a thiol group; R P4 It is a boric acid group or a borate ester group; R P5 It is a boric acid group or a borate ester group; R P6 It is protected by H or an amino group; R P5 'It is a boric acid group or a borate ester group; R P6 ' is a protecting group consisting of H or hydroxyl groups.

9. A pharmaceutical composition comprising the compound of any one of claims 1-7 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate thereof, and one or more pharmaceutically acceptable excipients.

10. The use of the compound of any one of claims 1-7 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof in the preparation of a medicament for the prevention and / or treatment of PKMYT1-related diseases; Preferably, the disease is selected from: tumors, inflammatory diseases, autoimmune diseases, infectious diseases, and precancerous syndromes; More preferably, the disease is a solid tumor, preferably selected from: colorectal cancer, gastric cancer, ovarian cancer, breast cancer, endometrial cancer, kidney cancer, prostate cancer, bladder cancer, liver cancer, lung cancer, esophageal cancer, pancreatic cancer, cervical cancer, glioblastoma, and neuroblastoma.

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