Compounds for inhibiting / inducing degradation of EGFR kinase and pharmaceutical compositions and applications thereof

Abstract

The present invention relates to a compound for inhibiting / inducing degradation of EGFR kinase, a pharmaceutical composition and application thereof. The compound is a compound shown in formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, or a hydrate thereof, or a solvate thereof, or a racemate thereof, or a polymorph thereof, or an isotopic variant thereof, or a metabolite thereof, or a prodrug thereof, wherein the R 1 ~R 3 and X 1 , L 1 ~L 4 , Y 1 , Y 2 , Y 3 , A, B, C, D and W groups are as defined in the specification. The compounds of the present invention and the pharmaceutical compositions containing the compounds can be used to treat diseases associated with EGFR kinase, such as cancer; the present invention also provides the preparation and use of the pharmaceutical compositions.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular, provides compounds capable of inhibiting EGFR kinase or inducing degradation of EGFR, and pharmaceutical compositions and applications thereof. Background Art

[0002] Lung cancer is one of the most common malignant tumors. In 2018, there were 2.1 million new cases of lung cancer worldwide, accounting for 11.6% of all new tumor cases; 1.8 million deaths, accounting for 18.4% of all tumor deaths. Among them, non-small cell lung cancer (NSCLC) accounts for about 80%-85% of the total number of lung cancers. Epidermal growth factor receptor (EGFR) is the most common driver gene for non-small cell lung cancer. About 50% of Chinese non-small cell lung cancer patients and 11-16% of non-small cell lung cancer patients in Western countries have EGFR gene mutations. The most common mutation types are exon 19 deletion mutation (del E746-A750) and exon 21 L858R point mutation, accounting for about 90% of all EGFR mutation populations.

[0003] EGFR small molecule inhibitors are the standard first-line treatment for non-small cell lung cancer with EGFR gene mutations and have been widely used in the field of lung cancer treatment. They competitively bind to EGFR with endogenous ligands, inhibit the activation of tyrosine kinase, and then block the EGFR signaling pathway, inhibiting the proliferation and metastasis of tumor cells and promoting apoptosis of tumor cells, among other biological effects.

[0004] The first-generation EGFR small molecule inhibitors Gefitinib and Erlotinib have been used to treat advanced non-small cell lung cancer with activating EGFR mutations (L858R.del E746-A750). However, patients will develop resistance to Gefitinib and Erlotinib after 10-12 months of use, and more than 50% of resistant patients are due to secondary mutations of EGFR T790M. The second-generation EGFR irreversible inhibitor Afatinib is effective for patients with advanced non-small cell lung cancer with activating EGFR mutations (L858R, del E746-A750), but it cannot solve the clinical resistance caused by EGFR T790M mutations, and Afatinib lacks selectivity for wild-type EGFR and has greater toxicity. The third-generation irreversible inhibitor Osimertinib overcomes EGFR T790M resistance and can effectively treat patients with advanced non-small cell lung cancer with EGFR T790M mutation resistance in clinical practice. Although Osimertinib has achieved great success in the clinical treatment of non-small cell lung cancer with EGFR T790M mutation, some patients who benefited from it developed drug resistance after 9-14 months of treatment (Nature Medicine, 2015, 21(6), 560-562). Studies have shown that up to 22% of drug-resistant patients are resistant to Osimertinib due to the EGFR C797S mutation (JAMA Oncol. 2018; 4(11): 1527-1534). The EGFR C797S mutation causes the cysteine ​​at position 797 to mutate to serine, and Osimertinib cannot covalently bind to EGFR, ultimately causing drug resistance. Currently, there is a lack of effective EGFR inhibitors for EGFR C797S alone in clinical practice. Therefore, the development of a new generation of EGFR inhibitors to meet clinical treatment needs is an urgent problem to be solved.

[0005] The Ubiquitin-Proteasome System (UPS) is a multi-component system for intracellular protein degradation, which is involved in important physiological and biochemical processes such as cell growth, differentiation, DNA replication and repair, cell metabolism, and immune response. Protein degradation mediated by the ubiquitin-proteasome pathway is an important mechanism for the body to regulate intracellular protein levels and functions, and plays an important role in maintaining protein homeostasis in the body. Inducing the degradation of EGFR through the intracellular ubiquitin-proteasome pathway provides a new approach for the treatment of non-small cell lung cancer. Summary of the invention

[0006] The purpose of the present invention is to overcome the problems existing in the prior art and provide a compound for inhibiting / inducing degradation of EGFR kinase and a pharmaceutical composition and application thereof.

[0007] In a first aspect, the present invention provides a compound as shown in Formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, racemate, polymorph, isotopic variant, metabolite or prodrug thereof,

[0008]

[0009] in,

[0010] Y 1 and Y 2 Independently selected from O, S, Se, NR 4 or CR 4 ;

[0011] Y 3 are independently selected from C or N;

[0012] X 1 are independently selected from C or N;

[0013] Ring A is independently selected from a 5-6 membered aromatic heterocyclic ring or a saturated heterocyclic ring, the aromatic heterocyclic ring or the saturated heterocyclic ring contains 1 to 3 heteroatoms, the heteroatoms are independently selected from one or more of N, O and S, and the aromatic heterocyclic ring is optionally substituted by n R 5 Substitution, wherein n is selected from any integer between 0 and 3;

[0014] The groups W and B are independently selected from a 5-15 membered aromatic ring or heteroaromatic ring, a heteroaromatic and heteroalkyl ring, a heteroaromatic and spiro ring, or a heteroaromatic and bridged ring, wherein each of the aromatic ring, heteroaromatic ring, heteroaromatic and heteroalkyl ring, heteroaromatic and spiro ring, or heteroaromatic and bridged ring is optionally substituted by at least one R 6 replace;

[0015] R 1 , R 2 and R 3 Each independently selected from hydrogen, deuterium, halogen, cyano, amino, hydroxyl, -NR 7 R 8 、-OR 7 , C 1 -C 6 Alkyl, C 1 -C 6 Heteroalkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 3 -C 8 Cycloalkyl, C 3 -C 8 Heterocycloalkyl, C 1 -C 3 Alkoxy, C1 -C 6 Haloalkoxy or C 1 -C 6 wherein the alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, haloalkoxy and haloalkyl are each optionally substituted with at least one R 9 replace;

[0016] R 4 , R 5 and R 6 independently selected from hydrogen, deuterium, halogen, cyano, amino, carbonyl, hydroxyl, H, C 3 -C 8 Cycloalkyl, C 3 -C 8 Heterocycloalkyl, C 1 -C 6 Alkyl, C 1 -C 6 Heteroalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkoxy, -NR 7 R 8 OR 7 , and wherein each of the cycloalkyl, heterocycloalkyl, alkyl, heteroalkyl, alkoxy and haloalkoxy groups is optionally substituted by at least one R 9 replace;

[0017] R 7 and R 8 independently selected from hydrogen, deuterium, C 1 -C 6 Alkyl, C 1 -C 6 Heteroalkyl, C 3 -C 8 Cycloalkyl or C 3 -C 8 wherein the alkyl, heteroalkyl, cycloalkyl and heterocycloalkyl are each optionally substituted by at least one R 9 replace;

[0018] L 1 , L 2 , L 3 and L 4 are independently selected from absence, self-bond, -O-, -(CH 2 ) m -C(O)-, -C(O)-C(H 2 ) p -、-C(O)-C(O)-、NR 10 -C(O)-, -C(O)-NR10 、-C(O)O-、-CH 2 -CF 2 -CH 2 -、-CH 2 -、 wherein each m and p is independently any integer from 0 to 3;

[0019] The groups C, D and E are each independently selected from the group consisting of 6 -C 10 aryl, 5-15 membered heteroaryl, 4-8 membered monoheterocycloalkyl, 6-15 membered spiroheterocycloalkyl, 6-15 membered bridged heterocycloalkyl or 6-15 membered and heterocycloalkyl, wherein each of the aryl, heteroaryl, monoheterocycloalkyl, spiroheterocycloalkyl, bridged heterocycloalkyl and and heterocycloalkyl is optionally substituted by at least one R 11 replace;

[0020] R 9 and R 10 are each independently selected from hydrogen, deuterium, C 1 -C 6 Alkyl, C 1 -C 6 Heteroalkyl, C 3 -C 8 Cycloalkyl or C 3 -C 8 wherein the alkyl, heteroalkyl, cycloalkyl and heterocycloalkyl are each optionally substituted by at least one R 12 replace;

[0021] R 11 Selected from hydrogen, deuterium, halogen, cyano, amino, hydroxyl, C 1 -C 6 Alkyl, C 1 -C 6 Heteroalkyl, C 3 -C 8 Cycloalkyl, C 3 -C 8 Heterocycloalkyl, C 1 -C 3 Alkoxy, C 1 -C 6 Haloalkoxy or C 1 -C 6 wherein the alkyl, heteroalkyl, alkoxy, haloalkoxy, haloalkyl, cycloalkyl and heterocycloalkyl are each optionally substituted with at least one R 12 replace;

[0022] R 1 To R 11The heteroatoms or heteroatomic groups contained in the heteroalkyl, heterocycloalkyl and heteroaryl groups are independently selected from -C(=O)N(R 12 )-、-N(R 12 )-, -NH-, -N=, -O-, -S-, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) 2 -or-N(R 12 )C(=O)N(R 12 )-, and the number of the heteroatoms or heteroatom groups is independently selected from 1, 2 and 3;

[0023] R 12 Each is independently selected from hydrogen, chlorine, fluorine, cyano, hydroxy, amino, isopropyl, cyclopropyl, methyl, ethyl, difluoromethyl, trifluoromethyl, methoxy, trifluoromethoxy, ethoxy, 2,2-difluoroethoxy, 2,2,2-trifluoroethoxy and phenyl.

[0024] Preferably, it is a compound as shown in formula I-1 or I-2,

[0025]

[0026] Preferably, the present invention provides a specific compound as shown in any one of Formula 1 to Formula 3, whose structural formula is:

[0027]

[0028]

[0029] Preferably, the stereoisomers include enantiomers or diastereomers.

[0030] In a second aspect, the present invention provides a pharmaceutical composition comprising an effective dose of one or more of the compounds of the present invention, their pharmaceutically acceptable salts, their hydrates, their solvates, their stereoisomers, their tautomers, their racemates, their polymorphs, their isotopic variants, their metabolites and their prodrugs.

[0031] Preferably, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient.

[0032] Preferably, the auxiliary material comprises a pharmaceutically acceptable excipient or therapeutic agent.

[0033] In another aspect, the present invention provides a kit comprising a compound of the present invention, and other therapeutic agents and a pharmaceutically acceptable carrier, adjuvant or vehicle.

[0034] In a third aspect, the present invention provides the use of one or more of the compounds of the present invention, their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, tautomers, racemates, polymorphs, isotopic variants, metabolites and prodrugs thereof in the preparation of drugs for treating and / or preventing EGER kinase-mediated diseases.

[0035] In a fourth aspect, the present invention provides the use of one or more of the compounds of the present invention, their pharmaceutically acceptable salts, their hydrates, their solvates, their stereoisomers, their tautomers, their racemates, their polymorphs, their isotopic variants, their metabolites and their prodrugs in the preparation of drugs for treating and / or preventing cancer.

[0036] In another aspect, the present invention provides a method for treating and / or preventing an EGFR kinase-mediated disease in a subject, comprising administering to the subject a compound or composition of the present invention.

[0037] In another aspect, the present invention provides a compound of the present invention or a composition of the present invention for use in the treatment and / or prevention of EGFR kinase-mediated diseases.

[0038] Compared with the prior art, the present invention has the following beneficial effects:

[0039] The present invention provides a class of compounds. It is proved by relevant cell activity tests that the compounds in the present invention have good activity in inhibiting the proliferation of Ba / F3 (EGFR L858R / T790M / C797S) cells, and the activities of multiple compounds are less than 100nM, showing extremely important anti-tumor potential. At the same time, as a Protac molecule, under the premise of a large molecular weight, it still has a good oral exposure, showing good PK properties, and can be administered orally. Therefore, the compounds provided by the present invention have good clinical application prospects and can be used to prepare anticancer drugs. DETAILED DESCRIPTION

[0040] In a specific embodiment, the diseases treated by the present invention include cancer, such as ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal stromal tumor (GIST), thyroid cancer, bile duct cancer, endometrial cancer, renal cancer, anaplastic large cell lymphoma, acute myeloid leukemia (AML), multiple myeloma, melanoma, mesothelioma.

[0041] General Terms and Definitions

[0042] Unless otherwise stated, the terms used in the present invention have the following meanings.

[0043] "Alkyl" refers to a saturated aliphatic hydrocarbon group, including straight and branched groups of 1 to 20 carbon atoms, for example, straight and branched groups of 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms or 1 to 4 carbon atoms. In the present invention, "alkyl" can be a monovalent, divalent or trivalent group. Non-limiting examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl and various branched chain isomers thereof, etc. Non-limiting examples also include, but are not limited to, methylene, methine, ethylene, ethylene, propylene, propylene, butylene, butylene and various branched chain isomers thereof. In addition, in the present invention, "alkyl" may be optionally substituted or unsubstituted.

[0044] "Alkoxy" refers to an "-O-alkyl" group wherein "alkyl" is as defined above.

[0045] "Alkenyl" refers to an unsaturated aliphatic hydrocarbon group, including straight and branched chain groups of 1 to 20 carbon atoms and at least one carbon-carbon double bond, for example, straight and branched chain groups of 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. In the present invention, "alkenyl" can be a monovalent, divalent, or trivalent group. Non-limiting examples include, but are not limited to, vinyl (-CH=CH 2 ), propen-1-yl (-CH=CH-CH 3 ), propene-2-yl (-C(CH 3 )=CH 2 ), buten-1-yl (-CH=CH-CH 2 -CH 3 ), butene-2-yl (-C (C 2 H 5 )=CH 2 ), 1-methylpropen-1-yl (-C(CH 3 )=CH-CH 3 ) and its various branched isomers, etc. Non-limiting examples also include, but are not limited to, 1,1-vinylene (=C=CH 2 ), 1,2-vinylene (-CH=CH-), 1,1-propenylene (=C=CH-CH 3 ), 1,2-propenyl (-CH=C(CH3 )-), 1,3-propenylene (-CH=CH-CH 2 -) and its various branched isomers. In addition, in the present invention, "alkenyl" may be optionally substituted or unsubstituted.

[0046] "Alkynyl" refers to an unsaturated aliphatic hydrocarbon group, including straight and branched groups of 1 to 20 carbon atoms and at least one carbon-carbon triple bond, for example, straight and branched groups of 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. In the present invention, "alkynyl" can be a monovalent, divalent, or trivalent group. Non-limiting examples include, but are not limited to, ethynyl (-C≡CH), propynyl (C≡C-CH 3 ), butynyl Pentynyl and its various branched chain isomers, etc. Non-limiting examples also include, but are not limited to, ethynyl (C≡C-), propynyl Butynylene and various branched isomers thereof. In addition, in the present invention, "alkynyl" may be optionally substituted or unsubstituted.

[0047] "Heteroalkyl" refers to a saturated aliphatic hydrocarbon group, including straight and branched chain groups of 2 to 20 atoms, for example, straight and branched chain groups of 2 to 18 atoms, 2 to 12 atoms, 2 to 8 atoms, 2 to 6 atoms or 2 to 4 atoms, wherein one or more atoms is selected from nitrogen, oxygen or S(O) m (wherein m is 0, 1 or 2) heteroatoms, and the rest are carbon. In the present invention, "heteroalkyl" can be a monovalent, divalent or trivalent group. Non-limiting examples include, but are not limited to, methoxymethyl (2-oxapropyl), methylthiomethyl (2-thiapropyl), methylaminomethyl (2-aziridine propyl) and various branched chain isomers thereof. In addition, in the present invention, "heteroalkyl" can be optionally substituted or unsubstituted.

[0048] "Cycloalkyl" refers to a saturated or partially unsaturated, monocyclic or polycyclic aliphatic hydrocarbon group, including 3 to 12 ring atoms, for example, 3 to 12, 3 to 10 or 3 to 6 ring atoms (i.e., 3 to 6 rings). Non-limiting examples of monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc. In the present invention, "cycloalkyl" can be optionally substituted or unsubstituted.

[0049] "Heterocycloalkyl" refers to a saturated or partially unsaturated, monocyclic or polycyclic aliphatic hydrocarbon group comprising 3 to 20 ring atoms, for example 3 to 16, 3 to 12, 3 to 10 or 3 to 6 ring atoms, wherein one or more of the ring atoms is selected from nitrogen, oxygen or S(O) m (wherein m is 0,1 or 2) heteroatoms, and the remaining ring atoms are carbon.Preferably, heterocycloalkyl includes 3 to 12 ring atoms, wherein 1 to 4 ring atoms are heteroatoms, more preferably includes 3 to 10 ring atoms, most preferably includes 5 or 6 ring atoms, wherein 1 to 4, preferably 1 to 3, more preferably 1 to 2 are heteroatoms.The limiting examples of monocyclic heterocycloalkyl include but are not limited to pyrrolidinyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl etc.The limiting examples of polycyclic heterocycloalkyl include but are not limited to the heterocycloalkyl of ring, spirocycle or bridged ring.

[0050] "Halogen" refers to fluorine, chlorine, bromine and iodine, with fluorine, chlorine and bromine being preferred.

[0051] "Haloalkyl" or "haloalkoxy" refers to an alkyl or alkoxy group substituted by one or more halogen atoms which are the same or different. Preferred examples of alkyl or alkoxy include, but are not limited to, trifluoromethyl, trifluoroethyl, and trifluoromethoxy.

[0052] "Cyano" refers to a "-CN" group.

[0053] "Hydroxy" refers to a "-OH" group.

[0054] "Amino" refers to "-NH 2 ” group.

[0055] "Carbamoyl" refers to "-(C=O)-NH 2 ” group.

[0056] "Aryl" refers to monocyclic, bicyclic and tricyclic carbon ring systems containing 6-14 ring atoms, wherein at least one ring system is aromatic, wherein each ring system contains a ring consisting of 3-7 atoms and has one or more points of attachment to the rest of the molecule. Examples include, but are not limited to, phenyl, naphthyl, anthracene, etc. Preferably, the aryl group is a carbon ring system of 6-10 or 6-7 ring atoms.

[0057] "Heteroaryl" refers to monocyclic, bicyclic and tricyclic ring systems containing 5-14 ring atoms, wherein at least one ring system is aromatic and at least one ring system contains one or more heteroatoms selected from nitrogen, oxygen and sulfur, wherein each ring system contains a ring consisting of 5-7 atoms and has one or more points of attachment to the rest of the molecule. The term "heteroaryl" can be used interchangeably with the terms "heteroaromatic ring" or "heteroaromatic compound". Examples include, but are not limited to: furanyl, imidazolyl, 2-pyridyl, 3-pyridyl, thiazolyl, purinyl, quinolyl. Preferably, the heteroaryl is a ring system of 5-10 ring atoms.

[0058] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0059] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are replaced independently of one another by a corresponding number of substituents.

[0060] "Pharmaceutically acceptable salt" refers to a salt prepared from a compound of the present invention and a relatively non-toxic acid or base. When the compound of the present invention contains a relatively acidic functional group (e.g., a carboxyl group or a sulfonic acid group), a base addition salt can be obtained by contacting its free form with a sufficient amount of a base in a pure solution or a suitable inert solvent. Non-limiting examples of pharmaceutically acceptable base addition salts include, but are not limited to, sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, organic amine salts, or similar salts. When the compound of the present invention contains a relatively alkaline functional group (e.g., an amino group or a guanidine group), an acid addition salt can be obtained by contacting its free form with a sufficient amount of an acid in a pure solution or a suitable inert solvent. Non-limiting examples of pharmaceutically acceptable acid addition salts include, but are not limited to, inorganic acid salts (e.g., hydrochlorides, hydrobromides, hydroiodides, nitrates, carbonates, bicarbonates, phosphates, monohydrogen phosphates, dihydrogen phosphates, phosphites, sulfates, hydrogen sulfates, etc.), organic acid salts (e.g., acetates, propionates, isobutyrates, malonates, succinates, suberates, maleates, fumarates, citrates, tartrates, lactates, mandelates, benzoates, phthalates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, glucuronic acid, etc.), and amino acid salts (e.g., arginine salts, etc.). Specific forms of pharmaceutically acceptable salts can also be found in Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66: 1-19). Certain specific compounds of the present invention contain basic and acidic functional groups and can be converted into either base addition salts or acid addition salts. Preferably, the salt is contacted with an alkali or acid in a conventional manner, and the parent compound is separated, thereby regenerating the neutral form of the compound. The parent form of the compound differs from its various salt forms in certain physical properties, such as different solubility in polar solvents. According to embodiments of the present invention, the pharmaceutically acceptable salt of the compound preferably as shown in Formula I is an acid addition salt, preferably hydrochloride, hydrobromide, phosphate or sulfate, more preferably hydrochloride.

[0061] A "pharmaceutical composition" refers to a composition for pharmaceutical use, which comprises one or more compounds as shown in Formula I or a pharmaceutically acceptable form thereof (e.g., salts, hydrates, solvates, stereoisomers, tautomers, racemates, polymorphs, isotopic variants, metabolites, prodrugs, etc.), and other components (e.g., pharmaceutically acceptable excipients).

[0062] In the present invention, "pharmaceutically acceptable excipients" refer to auxiliary materials widely used in the field of drug production. The main purpose of using excipients is to provide a pharmaceutical composition that is safe to use, stable in nature and / or has specific functionality, and also to provide a method so that after the drug is administered to the subject, the active ingredient can be dissolved at a desired rate, or to promote the effective absorption of the active ingredient in the subject receiving the drug. Pharmaceutically acceptable excipients can be inert fillers or functional ingredients that provide a certain function to the pharmaceutical composition (for example, stabilizing the overall pH value of the composition or preventing the degradation of the active ingredient in the composition). Non-limiting examples of pharmaceutically acceptable excipients include, but are not limited to, adhesives, suspending agents, emulsifiers, diluents (or fillers), granulating agents, adhesives, disintegrants, lubricants, anti-adhesive agents, glidants, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, sweeteners, etc.

[0063] The pharmaceutical compositions of the present invention can be prepared by any method known to those skilled in the art, for example, conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding and / or lyophilizing processes.

[0064] In the present invention, the purpose of using the pharmaceutical composition is to promote the administration of the organism, which is conducive to the absorption of the active ingredient and then exerts biological activity. The pharmaceutical composition of the present invention can be administered in any form, including injection (intra-arterial, intravenous, intramuscular, intraperitoneal, subcutaneous), mucosal, oral (oral solid preparations, oral liquid preparations), rectal, inhalation, implantation, local (e.g., eye) administration, etc. Non-limiting examples of oral solid preparations include, but are not limited to, powders, capsules, lozenges, granules, tablets, etc. Non-limiting examples of liquid preparations for oral or mucosal administration include, but are not limited to, suspensions, tinctures, elixirs, solutions, etc. Non-limiting examples of topical preparations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops or serum preparations. Non-limiting examples of parenteral preparations include, but are not limited to, solutions for injection, dry powders for injection, suspensions for injection, emulsions for injection, etc. The pharmaceutical composition of the present invention can also be made into controlled release or delayed release dosage forms (e.g., liposomes or microspheres).

[0065] Preferably, the compound of the present invention or the pharmaceutical composition comprising the same is administered orally or intravenously to an individual in need thereof. Depending on the specific circumstances of the subject to be administered, other routes of administration may also be applicable or even preferred. For example, for patients who are forgetful or irritable to oral medications, transdermal administration will be a very important route of administration. In the present invention, the route of administration can be changed or adjusted in any suitable manner to meet the needs of the nature of the drug, the convenience of the patient and medical staff, and other relevant factors.

[0066] The compound of the present invention or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug or pharmaceutical composition comprising the same has excellent PLK1 enzyme inhibitory activity and cell proliferation inhibitory activity, can be used as a PLK inhibitor for preventing and / or treating diseases or conditions caused by overexpression of PLK, and has good clinical application and medical use. Preferably, a non-limiting example of a disease or condition caused by overexpression of PLK1 is cancer, including but not limited to blood tumors, pancreatic cancer, colorectal cancer and lung cancer.

[0067] The technical scheme of the present invention will be described below in conjunction with specific embodiments. The following embodiments are provided to further illustrate the present invention, but are not intended to limit the scope of the present invention. It will be apparent to those skilled in the art that various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0068] The preparation of the compounds of the present invention can be achieved by synthetic methods well known to those skilled in the art, including but not limited to the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitution methods well known to those skilled in the art, and preferred embodiments include but are not limited to the embodiments of the present invention. The known starting materials used in the present invention can be synthesized by methods known in the art, or purchased by conventional commercial means (for example, purchased from Shaoyuan Chemical Technology, Beijing Coupling Technology, etc.). Unless otherwise specified, the reactions are carried out under an argon atmosphere or a nitrogen atmosphere. The hydrogenation reaction is usually evacuated, filled with hydrogen, and the operation is repeated 3 times. The reaction temperature is room temperature, and the temperature range is 20°C-30°C. Monitoring of the reaction progress can be achieved by synthetic methods well known to those skilled in the art, including but not limited to thin layer chromatography (TLC). The thin layer chromatography silica gel plate uses Qingdao Ocean GF254 silica gel plate, and the developing solvent system includes but is not limited to A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system, and the volume ratio of the solvent can be adjusted according to the polarity of the compound.

[0069] The separation and purification of the compounds of the present invention can be achieved by synthetic methods well known to those skilled in the art, including but not limited to column chromatography (CC), high performance liquid chromatography (HPLC), ultra-high performance liquid chromatography (UPLC), etc. Column chromatography generally uses Qingdao Ocean 200-300 mesh silica gel as a carrier, and the eluent system includes but is not limited to A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvent can be adjusted according to the polarity of the compound, and a small amount of acidic or alkaline anti-tailing reagent can also be added for adjustment. The HPLC spectrum is measured using an Agilent 1200DAD HPLC chromatograph (chromatographic column: Sunfire C18, 150×4.6mm, 5μm) or a Waters 2695-2996 HPLC chromatograph (chromatographic column: Gimini C18, 150×4.6mm, 5μm).

[0070] The structural identification of the compounds of the present invention can be achieved by methods well known to those skilled in the art, including but not limited to nuclear magnetic resonance (NMR), mass spectrometry (MS), etc. The NMR spectrum is measured using a Bruker AVANCE-400 or Varian Oxford-300 NMR spectrometer, and the measurement solvent is deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated chloroform (CDC1 3 ) or deuterated methanol (CD 3 OD), the internal standard was tetramethylsilane (TMS), and the chemical shift was 10 -6 The MS spectra were measured using an Agilent SQD (ESI) mass spectrometer (model: 6110) or a Shimadzu SQD (ESI) mass spectrometer (model: 2020).

[0071] Preparation of intermediates

[0072] Preparation of intermediate INT-1

[0073]

[0074] Preparation method:

[0075] Step 1: Synthesis of compound INT-1B

[0076] Compound INT-1A (100.0 g, 457 mmol) was dissolved in 1,4-dioxane (1000 mL), and 85% hydrazine hydrate (51.6 g, 914 mmol) was added to the reaction solution, and then the reaction solution was heated to 95°C and stirred for 16 hours. After TLC showed that the reaction was completed, the reaction solution was cooled to room temperature, and water (3 L) was added to the reaction solution with stirring, and a large amount of white solid precipitated. After stirring for 30 minutes, it was filtered, and the filter cake was washed with water (500 mL), and then the filter cake was dried to obtain compound INT-1B (88.0 g, white solid, yield 89.8%).

[0077] MS (ESI): m / z 215.1 [M+1] + / 217.1[M+3] + .

[0078] Step 2: Synthesis of compound INT-1C

[0079] Compound INT-1B (70.0 g, 327 mmol) was dissolved in N, N-dimethylacetamide (700 mL), ethyl bromoacetate (108.6 g, 654 mmol) was added to the reaction solution, and then the reaction solution was heated to 100°C and stirred for 40 hours. After TLC showed that the reaction was completed, the reaction solution was poured into a mixed solvent of ethyl acetate (1000 mL) and water (800 mL), the organic phase was separated, the aqueous phase was extracted with ethyl acetate (300 mL×2), the combined organic phase was washed with saturated sodium chloride solution (500 mL), the organic phase was separated, concentrated, and the residue was purified by chromatography on a silica gel column (eluent: petroleum ether / ethyl acetate = 10 / 1 (volume ratio)) to obtain compound INT-1C (50.0 g, white solid, yield 51.0%).

[0080] MS (ESI): m / z 301.1 [M+1] + / 303.1[M+3] + .

[0081] Step 3: Synthesis of compound INT-1E

[0082] The compound pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester (22.2 g, 103 mmol) was dissolved in anhydrous tetrahydrofuran (100 mL), and N,N'-carbonyldiimidazole (16.7 g, 103 mmol) was added in batches at 0°C. After the addition, the mixture was transferred to 25°C and stirred for 16 hours to obtain a solution of INT-1D. The compound INT-1C (20.0 g, 66.7 mmol) was dissolved in anhydrous tetrahydrofuran (100 mL), cooled to -70°C, and the reaction liquid temperature was controlled at -60°C. A 2M tetrahydrofuran solution of lithium diisopropylamide (51.5 mL, 103 mmol) was added dropwise. After the addition, the mixture was reacted at -70°C for 30 minutes, and then the solution of INT-1D was added dropwise to the reaction liquid, and the reaction liquid temperature was controlled below -60°C. After the addition, the mixture was naturally heated to 25°C and stirred for 2 hours. After TLC showed that the reaction was completed, saturated ammonium chloride solution (100 mL) and ethyl acetate (100 mL) were added to the reaction solution, the aqueous phase was extracted with ethyl acetate (50 mL×3), and then the organic phases were combined and concentrated. The residue was quickly washed with a silica gel column (eluent: petroleum ether / ethyl acetate = 1 / 1 (volume ratio)) to give a crude compound INT-1E (33.1 g, brown oil, yield 100%).

[0083] MS (ESI): m / z 498.1 [M+1] + / 500.1[M+3] + .

[0084] Step 4: Synthesis of compound INT-1F

[0085] Compound INT-1E (33.1 g, 66.7 mmol) was dissolved in ethyl acetate (300 mL), the reaction solution was cooled to 0°C, 2M hydrochloric acid / ethyl acetate solution (600 mL) was added dropwise, and the mixture was stirred at 40°C for 16 hours. After TLC showed that the reaction was complete, the reaction solution was concentrated to dryness to obtain compound INT-1F (26.5 g, brown oil, yield 100%).

[0086] MS (ESI): m / z 398.0 [M+1] + / 400.0[M+3] + .

[0087] Step 5: Synthesis of compound INT-1G

[0088] Compound INT-1F (26.5 g, 66.7 mmol) was dissolved in water (300 mL) and tert-butyl alcohol (100 mL), and potassium thiocyanate (9.8 g, 100 mmol) was added to the reaction solution. After the addition, the temperature was raised to 90°C for reaction for 3 hours. After TLC showed that the reaction was completed, ethyl acetate (300 mL) was poured into the reaction solution, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (60 mL×2), and then the organic phases were combined and concentrated. The residue was purified by chromatography on a silica gel column (eluent: petroleum ether / ethyl acetate = 1 / 1 (volume ratio)) to obtain compound INT-1G (6.8 g, brown foamy solid, three-step yield 23.1%).

[0089] MS (ESI): m / z 439.0 [M+1] + / 441.0[M+3] + .

[0090] Step 6: Synthesis of compound INT-1H

[0091] Compound INT-1G (6.8 g, 15.5 mmol) was dissolved in glacial acetic acid (68 mL), the reaction solution was cooled to 0°C, 30% hydrogen peroxide (7.03 g, 62 mmol) was added dropwise at a temperature below 10°C, and the reaction solution was transferred to 25°C and stirred for 1 hour. After TLC showed that the reaction was completed, ethyl acetate (100 mL) was added, and then the pH was adjusted to 7-8 with a saturated sodium carbonate solution, the organic layer was separated, the aqueous layer was extracted with ethyl acetate (50 mL×2), the organic phases were combined and concentrated, and the residue was purified by chromatography on a silica gel column (eluent: ethyl acetate = 1 / 1 (volume ratio)) to obtain compound INT-1H (4.2 g, brown foamy solid, yield 66.7%).

[0092] MS (ESI): m / z 407.0 [M+1] + / 409.0[M+3] + .

[0093] Step 7: Synthesis of compound INT-1J

[0094] Compound INT-1H (4.2 g, 10.3 mmol) was dissolved in tetrahydrofuran (40 mL) and water (5 mL), the reaction solution was cooled to 0°C, lithium hydroxide monohydrate (0.32 g, 13.4 mmol) was added to the reaction solution, and after the addition, the reaction solution was transferred to 25°C for reaction for 1 hour. After TLC showed that the reaction was completed, the pH was adjusted to 6-7 with 4M hydrochloric acid, and then the reaction solution was concentrated to dryness to obtain compound INT-1J (4.0 g, brown foamy solid, yield 100%).

[0095] MS (ESI): m / z 379.0 [M+1] + / 381.0[M+3] + .

[0096] Step 8: Synthesis of compound INT-1

[0097] Compound INT-1J (4.0 g, 10.3 mmol) was dissolved in N,N-dimethylformamide (50 mL), and 2-aminothiazole (1.34 g, 13.4 mmol) and N,N-diisopropylethylamine (4.0 g, 30.9 mmol) were added to the reaction solution in sequence. The reaction solution was then cooled to 0°C, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (5.9 g, 15.4 mmol) was added in batches. After the addition was complete, the mixture was transferred to 25°C and stirred for reaction for 2 hours. After TLC showed that the reaction was completed, the reaction solution was poured into ethyl acetate (100 mL) and water (120 mL), the organic phase was separated, the aqueous phase was extracted with ethyl acetate (50 mL×2), and then the organic phases were combined, washed with saturated sodium chloride solution (40 mL×2), the organic phase was separated and concentrated, and the residue was purified by silica gel column (eluent: ethyl acetate / methanol = 10 / 1 (volume ratio)) chromatography to obtain compound INT-1 (2.8 g, off-white solid, yield 58.9%).

[0098] MS (ESI): m / z 461.0 [M+1] + / 463.0[M+3] + .

[0099] 1 H NMR (300 MHz, DMSO-d 6 )δ12.83(s,1H),8.36(s,1H),7.76(t,J=1.0Hz,1H),7.67(s,1H),7.50(d,J=3.6Hz,1H),7.28(d,J= 3.6Hz,1H),7.07(dd,J=9.9,1.2Hz,1H),6.68(s,1H),4.02-3.96(m,2H),2.87(s,2H),2.71(s,2H).

[0100] Preparation of intermediate INT-2

[0101]

[0102] Preparation method:

[0103] Step 1: Synthesis of compound INT-2B

[0104] Concentrated sulfuric acid (103.0 g, 1.03 mol) was added dropwise to water (19 mL) at 0°C, stirred for 5 minutes, then warmed to room temperature, compound INT-2A (10.0 g, 57.8 mmol) was added, and after the addition, ethyl 4-chloroacetoacetate (12.4 g, 75.1 mmol) was added dropwise under an ice bath, and after the addition, the reaction solution was reacted at room temperature for 16 hours. After TLC showed that the reaction was completed, the reaction solution was slowly poured into ice water (500 mL), and a large amount of solid precipitated, filtered, and the filter cake was washed with water (50 mL) and dried to obtain compound INT-2B (10.0 g, white solid, yield 62.8%).

[0105] MS (ESI): m / z 274.9 [M+1] + / 276.9[M+3] + .

[0106] Step 2: Synthesis of compound INT-2C

[0107] Compound INT-2B (10.0 g, 36.3 mmol) was dissolved in water (100 mL), and then sodium hydroxide (4.79 g, 120 mmol) was added in batches. After the addition, the mixture was heated to 80°C and stirred for 5 hours. After TLC showed that the reaction was completed, the mixture was cooled to room temperature, and then the pH was adjusted to 5-6 with 1 M hydrochloric acid in an ice bath. A large amount of solid precipitated, which was filtered, and the filter cake was washed with water (50 mL) and dried to obtain compound INT-2C (6.0 g, white solid, yield 64.8%).

[0108] MS (ESI): m / z 255.0 [M+1] + / 257.0[M+3] + .

[0109] Step 3: Synthesis of compound INT-2D

[0110] Compound INT-2C (6.0 g, 23.5 mmol) was dissolved in anhydrous ethanol (60 mL), and concentrated sulfuric acid (1.2 g, 11.8 mmol) was then added dropwise under an ice bath. After the addition was complete, the reaction solution was heated to 80°C and stirred for 2 hours. After TLC showed that the reaction was complete, ethanol was evaporated under reduced pressure, the residue was dissolved in ethyl acetate (200 mL), and then water (200 mL) was added to separate the organic phase, the aqueous phase was extracted with ethyl acetate (100 mL), the organic phases were combined, dried, and spin-dried, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 10 / 1 (volume ratio)) to obtain compound INT-2D (6.0 g, colorless oil, yield 90.1%).

[0111] MS (ESI): m / z 283.0 [M+1]+ / 285.0[M+3] + .

[0112] Step 4: Synthesis of compound INT-2E

[0113] Compound INT-2D (3.0 g, 10.6 mmol) was dissolved in anhydrous tetrahydrofuran (30 mL), and acrylamide (7.53 g, 106 mmol) and potassium tert-butoxide (2.97 g, 26.5 mmol) were added in batches under an ice bath, and reacted at 25°C for 1 hour after the addition. After TLC showed that the reaction was completed, water (100 mL) was added to the reaction solution for dilution, and then extracted with ethyl acetate (100 mL×4), the organic phases were combined, dried, and spin-dried, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (volume ratio)) to obtain compound INT-2E (1.3 g, yellow solid, yield 39.8%).

[0114] MS (ESI): m / z 308.0 [M+1] + / 310.0[M+3] + .

[0115] 1 H NMR (300 MHz, DMSO-d 6 )δ10.92(s,1H),7.92(d,J=14.0Hz,2H),7.56(d,J=8.3Hz,1H),7.41(d,J=8.2Hz,1H),4.14(dd, J=12.0,4.7Hz,1H),2.77-2.67(m,1H),2.59-2.53(m,1H),2.38-2.24(m,1H),2.12-2.06(m,1H).

[0116] Step 5: Synthesis of compound INT-2F

[0117] The compound INT-2E (1.0 g, 3.25 mmol) was dissolved in a mixed solvent of 1,4-dioxane (8 mL) and water (2 mL), and N-benzyloxycarbonyl-3,6-dihydro-2H-pyridine-4-boronic acid pinacol ester (1.67 g, 4.87 mmol), cesium fluoride (1.0 g, 6.50 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (100 mg, 0.163 mmol) were added respectively. After the addition, the reaction solution was replaced with nitrogen three times, and then heated to 90 ° C under nitrogen protection for 16 hours. After TLC showed that the reaction was completed, the reaction solution was cooled to room temperature, diluted with water (80 mL), extracted with ethyl acetate (70 mL×3), the organic phases were combined, dried, and spin-dried. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (volume ratio)) to give compound INT-2F (800 mg, yellow solid, yield 55.5%).

[0118] MS (ESI): m / z 445.2 [M+1] + .

[0119] Step 6: Synthesis of compound INT-2G

[0120] Compound INT-2F (700 mg, 1.57 mmol) was dissolved in glacial acetic acid (15 mL), and 10% wet Pd / C (150 mg) was added. After the addition, the reaction system was replaced with hydrogen three times, and then the temperature was raised to 50° C. under a hydrogen pressure of 50 psi (hydrogen balloon) and stirred for 2 hours. After TLC showed that the reaction was complete, the reaction solution was cooled to room temperature, filtered, the filter cake was washed with methanol, the filtrate was collected, and dried to obtain compound INT-2G (450 mg, yellow oil, yield 91.5%).

[0121] MS (ESI): m / z 313.1 [M+1] + .

[0122] Step 7: Synthesis of compound INT-2H

[0123] Compound INT-2G (450 mg, 1.44 mmol) was dissolved in N, N-dimethylformamide (5 mL), and then tert-butyl bromoacetate (365 mg, 1.87 mmol) and N, N-diisopropylethylamine (945 mg, 4.32 mmol) were added, and the mixture was stirred at 25°C for 1 hour. After TLC showed that the reaction was completed, water (50 mL) was added to dilute the reaction solution, and then extracted with ethyl acetate (50 mL×2), the organic phases were combined, washed with saturated brine (30 mL×2), dried, and spin-dried, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1 (volume ratio)) to obtain compound INT-2H (400 mg, off-white solid, yield 65.1%).

[0124] MS (ESI): m / z 427.2 [M+1] + .

[0125] 1 H NMR (300 MHz, DMSO-d 6 )δ10.88(s,1H),7.82(s,1H),7.51-7.37(m,2H),7.13(d,J=8.2Hz,1H),4.09(dd,J=11.9,4.9Hz,1H),3.12(s,2H),2.92(d, J=11.1Hz,2H),2.76-2.67(m,1H),2.58-2.52(m,2H),2.33-2.24(m,3H),2.14-2.03(m,1H),1.73-1.70(m,4H),1.41(s,9H).

[0126] Step 8: Synthesis of compound INT-2

[0127] Compound INT-2H (100 mg, 0.234 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (0.33 mL) was added. After the addition was complete, the temperature was raised to 40°C and the reaction was allowed to proceed for 5 hours. After TLC showed that the reaction was complete, the reaction solution was dried to obtain compound INT-2 (80 mg, 0.216 mmol, light yellow foamy solid, yield 92.1%), which was directly used in the next step.

[0128] MS (ESI): m / z 371.1 [M+1] + .

[0129] Preparation of intermediate INT-3

[0130]

[0131] Preparation method:

[0132] Step 1: Synthesis of compound INT-3H

[0133] Diisopropylamine (2.2 mL, 15.6 mmol) was dissolved in anhydrous tetrahydrofuran (12.5 mL), and the reaction solution was cooled to 0°C, the reaction solution temperature was controlled at 0-5°C, 2.5 M n-butyllithium solution (6.25 mL, 15.6 mmol) was added dropwise, and the reaction solution temperature was maintained at 0-5°C for 15 minutes after the dropwise addition. The reaction solution was then cooled to -78°C, the reaction solution temperature was controlled below -60°C, compound INT-3G (2.83 g, 12.0 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) and added dropwise to the reaction solution, and then the reaction was carried out at -78°C for 1 hour. Finally, the reaction solution temperature was controlled below -60°C, N-phenylbis(trifluoromethanesulfonimide) was dissolved in anhydrous tetrahydrofuran (15 mL) and added dropwise to the reaction solution, and the reaction was carried out at -78°C for 2 hours after the dropwise addition, and then the temperature was raised to 25°C for 16 hours. After TLC showed that the reaction was completed, the reaction solution was concentrated, and then methyl tert-butyl ether (100 mL) was added to dissolve it, and it was washed with water (50 mL), 2M sodium hydroxide solution (50 mL×3), water (50 mL) and saturated saline solution (50 mL) in sequence. The organic phase was concentrated, dried, and spin-dried. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 10 / 1 (volume ratio)) to give compound INT-3H (1.3 g, light yellow oil, yield 29.7%).

[0134] MS (ESI): m / z 368 [M+1] + .

[0135] Step 2: Synthesis of compound 3B

[0136] Compound INT-2E (800 mg, 2.60 mmol) was dissolved in 1,4-dioxane (1 mL), and then diboric acid pinacol ester (1.0 g, 3.90 mmol), potassium acetate (780 mg, 7.80 mmol), [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (160 mg, 0.13 mmol) were added in sequence. After the addition, the reaction solution was replaced with nitrogen three times, and then heated to 90 ° C under nitrogen protection for 5 hours. After TLC showed that the reaction was completed, water (100 mL) was added to the reaction system for dilution, and ethyl acetate (50 mL × 3) was used for extraction. The organic phases were combined, dried, and spin-dried. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (volume ratio)) to obtain compound INT-3B (800 mg, brown oil, yield 86.8%).

[0137] MS (ESI): m / z 356.2 [M+1] + .

[0138] Step 3: Synthesis of compound INT-3C

[0139] Compound INT-3B (500 mg, 1.41 mmol) was dissolved in water (100 mL), and then compound INT-3H (1.5 g, 4.23 mmol), sodium carbonate (350 mg, 4.23 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (100 mg, 0.071 mmol) were added in sequence. After the addition, the reaction solution was replaced with nitrogen three times, and then heated to 55 ° C under nitrogen protection for 5 hours. After TLC showed that the reaction was completed, it was cooled to room temperature, and then water (100 mL) was added to dilute it, extracted with ethyl acetate (40 mL×3), the organic phases were combined, dried, and spin-dried. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (volume ratio)) to obtain compound INT-3C (500 mg, yellow solid, yield 79.6%).

[0140] MS (ESI): m / z 447.2 [M+1] + .

[0141] Step 4: Synthesis of compound INT-3D

[0142] Compound INT-3C (500 mg, 1.12 mmol) was dissolved in methanol (10 mL), and then 10% wet Pd / C (50 mg) was added. After the addition, the reaction system was replaced with hydrogen three times, and then reacted at 25° C. for 16 hours under a hydrogen pressure of 50 psi (hydrogen balloon). After TLC showed that the reaction was complete, the reaction solution was filtered and dried to obtain compound INT-3D (500 mg, yellow oil, yield 99.6%).

[0143] MS (ESI): m / z 449.2 [M+1] + .

[0144] Step 5: Synthesis of compound INT-3E

[0145] 8% hydrochloric acid / ethyl acetate solution (10 mL) was added to compound INT-3D (500 mg, 1.11 mmol), and the mixture was reacted at 25° C. for 2 hours. After TLC showed that the reaction was complete, the reaction solution was directly spin-dried to obtain a crude compound INT-3E which was directly used in the next step (500 mg, yellow solid, yield 100.0%).

[0146] MS (ESI): m / z 349.1 [M+1] + .

[0147] Step 6: Synthesis of compound INT-3F

[0148] Compound INT-3E (500 mg, 1.44 mmol) was dissolved in N,N-dimethylformamide (5 mL), and then tert-butyl bromoacetate (364 mg, 1.87 mmol) and N,N-diisopropylethylamine (945 mg, 4.32 mmol) were added, and the mixture was reacted at 25°C for 1 hour. After TLC showed that the reaction was completed, water (30 mL) was added to dilute the reaction solution, and then extracted with ethyl acetate (30 mL×2), the organic phases were combined, washed with saturated brine (20 mL×2), dried, and spin-dried, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1 (volume ratio)) to obtain compound INT-3F (600 mg, off-white solid, yield 96.2%).

[0149] MS (ESI): m / z 463.2 [M+1] + .

[0150] 1 H NMR (300 MHz, DMSO-d 6 )δ10.89(s,1H),7.87(s,1H),7.51(d,J=8.2Hz,2H),7.19(d,J=8.3Hz,1H),4.11(dd,J=11.9,4.8Hz,1H),3.30-3.26(m,2H),3.23-3. 13(m,2H),2.97-2.93(m,1H),2.87(s,1H),2.77-2.68(m,2H),2.60-2.53(m,2H),2.39-2.25(m,1H),2.15-2.08(m,2H),1.43(s,9H).

[0151] Step 7: Synthesis of compound INT-3

[0152] Compound INT-3F (100 mg, 0.234 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (0.33 mL) was added, and the temperature was raised to 40°C for reaction for 5 hours. After TLC showed that the reaction was complete, the reaction solution was spin-dried to obtain compound INT-3 (80 mg, light yellow foamy solid, yield 85.5%).

[0153] MS (ESI): m / z 407.1 [M+1] + .

[0154] Preparation and functional verification of target compounds

[0155] Example 1: Preparation of Compound 1

[0156] The structural formula of compound 1 is:

[0157]

[0158] The synthetic route of compound 1 is:

[0159]

[0160] The specific preparation method of compound 1 includes:

[0161] Preparation method:

[0162] Step 1: Synthesis of compound 1B

[0163] Compound 1A (2 g, 6.94 mmol) was dissolved in toluene (60 mL), and 4-bromoiodobenzene (4.92 g, 17.36 mmol), sodium tert-butoxide (6.6 g, 69.4 mmol), [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (340 mg, 0.42 mmol) were added in sequence at room temperature (20 ° C). After the reaction system was replaced with nitrogen three times, the temperature was raised to 90 ° C for 16 hours. After TLC showed that the reaction was completed, the reaction solution was poured into 100 mL of saturated ammonium chloride aqueous solution, extracted with ethyl acetate (70 mL × 3), and the organic layer was separated. The combined organic phase was washed with saturated brine (2 × 30 mL), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the organic solvent was dried by spin drying. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5:1 (volume ratio)) to obtain compound 1B (2 g, light yellow solid, yield 81.9%).

[0164] MS (ESI): m / z 353.1 [M+H] + .

[0165] Step 2: Synthesis of Compound 1C

[0166] Compound 1B (1.7 g, 4.83 mmol) was dissolved in dioxane (60 mL), and pinacol borate (2.0 g, 7.73 mmol), potassium acetate (1.4 g, 14.5 mmol), [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (350 mg, 0.44 mmol) were added in sequence at room temperature (20°C). The reaction system was replaced with nitrogen three times, and then heated to 90°C for 16 hours. After TLC showed that the reaction was completed, the reaction solution was poured into 100 mL of saturated aqueous ammonium chloride solution, extracted with ethyl acetate (60 mL×3), and the organic layer was separated. The combined organic phases were washed with saturated brine (2×30 mL), dried over anhydrous sodium sulfate, and the desiccant was removed by filtration. The organic solvent was dried and the obtained residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5:1 (volume ratio)) to obtain compound 1C (1.3 g, off-white solid, yield 67.3%).

[0167] MS (ESI): m / z 401.3 [M+H] + .

[0168] Step 3: Synthesis of Compound 1D

[0169] Compound 1C (566.8 mg, 1.42 mmol) and intermediate INT-1 (500.0 mg, 1.09 mmol) were added to a mixed solvent of 1,4-dioxane (6 mL) and water (1 mL), and sodium carbonate (231.1 mg, 2.18 mmol), 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (93.4 mg, 0.22 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (94.3 mg, 0.13 mmol) were added in sequence at room temperature (20°C). The reaction system was subjected to microwave reaction at 80°C for 2 hours under a nitrogen atmosphere. After the reaction of the raw material was completed as monitored by TLC, saturated aqueous ammonium chloride solution (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (40 mL×3). The organic phases were combined, washed with saturated brine (2×20 mL), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dried under reduced pressure. The residue was purified by HPLC (Waters Sunfire OBD 100x30 mm, 5 μm, mobile phase A: 0.1% TFA in water, mobile phase B: acetonitrile, gradient: 10% acetonitrile for 1 min, 52%-52% acetonitrile for 10 min, 95% acetonitrile for 14 min, and 10% acetonitrile for 16 min) to obtain compound 1D (30 mg, white solid, yield 4.2%).

[0170] MS (ESI): m / z: 328.1 [1 / 2M+H] + .

[0171] Step 4: Synthesis of Compound 1E

[0172] Compound 1D (30 mg, 0.046 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (0.4 mL) was added, and the mixture was reacted at 25° C. for 30 minutes. After TLC showed that the reaction was complete, the reaction solution was directly spin-dried to obtain crude compound 1E which was directly used in the next reaction (30 mg, yellow oil, 100.0%).

[0173] MS (ESI): m / z 555.2 [M+1] + .

[0174] Step 5: Synthesis of Compound 1

[0175] The intermediate INT-3 (28 mg, 0.071 mmol) was dissolved in N,N-dimethylformamide (0.5 mL), and N,N-diisopropylethylamine (30.3 mg, 0.24 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (29.8 mg, 0.078 mmol) were added to the reaction solution in sequence. After the addition, the mixture was reacted at 25°C for 15 minutes. Then, 1E (30 mg, 0.047 mmol) was dissolved in N,N-dimethylformamide (0.5 mL) and added dropwise to the reaction solution. After the addition, the mixture was stirred and reacted at 25°C for 1 hour. After TLC showed that the reaction was complete, the reaction solution was directly purified by HPLC (Waters Sunfire OBD 100x30 mm, 5μm, mobile phase A: 0.1% TFA in water, mobile phase B: acetonitrile, gradient: 10% acetonitrile for 1 min, 52%-52% acetonitrile for 10 min, 95% acetonitrile for 14 min, and 10% acetonitrile for 16 min) to obtain compound 1 (18 mg, off-white solid, yield 41.3%).

[0176] MS (ESI): m / z 472.2 [1 / 2M+1] + .

[0177] 1H NMR(600MHz,dmso)δ12.79(s,1H),10.87(s,1H),8.22(s,1H),7.86(s,1H),7 .65(s,1H),7.57(d,J=8.5Hz,2H),7.49(dd,J=11.2,5.9Hz,2H),7.25(d,J=3. 2Hz,1H),7.18(d,J=8.3Hz,1H),7.08(d,J=12.3Hz,1H),6.66(s,1H),6.51(d, J=8.7Hz,2H),4.40(s,2H),4.09(dt,J=16.4,6.4Hz,2H),4.05-3.90(m,4H),3 .27(s,2H),3.16(dt,J=25.7,8.4Hz,3H),2.94(d,J=10.0Hz,1H),2.83-2.77 (m,1H),2.76-2.64(m,2H),2.63-2.50(m,3H),2.45-2.38(m,2H),2.34-2.26( m,1H),2.20-2.12(m,1H),2.11-2.04(m,1H),2.00-1.92(m,1H),1.79(ddd,J= 11.8,8.8,7.0Hz,1H),1.47-1.39(m,1H),1.27(ddd,J=11.1,7.8,5.2Hz,1H).

[0178] Example 2: Preparation of Compound 2

[0179] The structural formula of compound 2 is:

[0180]

[0181] The synthetic route of compound 2 is:

[0182]

[0183] The specific preparation method of compound 2 includes:

[0184] Preparation method:

[0185] Step 1: Synthesis of compound 2B

[0186] Compound 2A (3 g, 13.95 mmol) was dissolved in a mixed solvent of tetrahydrofuran and triethylamine (75 mL / 7.5 mL), and trimethylsilyl acetylene (4.1 g, 41.85 mmol), bis(triphenylphosphine)palladium dichloride (369.6 mg, 0.84 mmol), and cuprous iodide (159.6 mg, 0.84 mmol) were added in sequence at room temperature (25°C). The reaction system was replaced with nitrogen three times, and then the temperature was raised to 60°C for 16 hours. After TLC showed that the reaction was completed, the reaction solution was poured into 100 mL of saturated aqueous ammonium chloride solution, extracted with ethyl acetate (60 mL×3), and the organic layer was separated. The combined organic phases were washed with saturated brine (2×30 mL), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the organic solvent was dried by spin drying. The obtained residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3:1 (volume ratio)) to obtain compound 2B (2.6 g, yellow solid, yield 80.0%).

[0187] MS (ESI): m / z: 234.1 [M+H] + .

[0188] Step 2: Synthesis of Compound 2C

[0189] 2B (2.5 g, 10.72 mmol) was dissolved in a mixed solvent of methanol and dichloromethane (30 mL / 30 mL), potassium fluoride (1.86 g, 32.16 mmol) was added at room temperature (25 ° C), and the reaction system was reacted at room temperature for 2 hours. After TLC showed that the reaction was completed, the reaction solution was poured into 100 mL of water, extracted with dichloromethane (40 mL × 3), and the organic layer was separated. The combined organic phase was washed with saturated brine (2 × 30 mL), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the organic solvent was spin-dried. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3:1 (volume ratio)) to obtain 2C (1.5 g, yellow solid, yield 86.9%).

[0190] MS (ESI): m / z: 162.1 [M+H] + .

[0191] Step 3: Synthesis of Compound 2D

[0192] Compound 2C (700 mg, 4.35 mmol) was dissolved in a mixed solvent of tetrahydrofuran and water (5 mL / 5 mL), and lithium hydroxide monohydrate (365 mg, 8.7 mmol) was added at room temperature (20°C), and stirred at room temperature for 5 hours. After TLC showed that the reaction was complete, the reaction solution was concentrated to remove the organic phase, and 10 mL of water was added to dilute it. The aqueous phase was adjusted to pH = 4 with 4M hydrochloric acid, and extracted with ethyl acetate (50 mL×3), and the organic layer was separated. The combined organic phase was washed with saturated brine (2×30 mL), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the organic solvent was spin-dried to obtain a crude product 2D (500 mg, white solid, yield 78.2%).

[0193] MS (ESI): m / z: 148.0 [M+H] + .

[0194] Step 4: Synthesis of compound 2E

[0195] Compound 2D (450 mg, 3.06 mmol) was added to N, N-dimethylformamide (10 mL), and N, N-diisopropyl-N-ethyl (792.0 mg, 6.12 mmol), 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (1.4 g, 3.67 mmol), and compound 2D-1 (735 mg, 3.67 mmol) were added at 0°C, and stirred for 16 hours. After TLC monitoring, the reaction solution was added with saturated aqueous ammonium chloride (50 mL) and extracted with ethyl acetate (40 mL × 3). After the organic phases were combined, they were washed with saturated brine (2 × 20 mL), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dried under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3:1 (volume ratio)) to obtain compound 2E (800 mg, light yellow solid, yield 79.5%).

[0196] MS (ESI): m / z: 330.2 [M+H] + .

[0197] Step 5: Synthesis of Compound 2F

[0198] Compound 2E (429.1 mg, 1.30 mmol) and intermediate INT-1 (500.0 mg, 1.09 mmol) were added to N, N-dimethylformamide (10 mL), and N, N-diisopropyl-N-ethyl (154.6 mg, 1.20 mmol), cuprous iodide (10.3 mg, 0.055 mmol), triphenylphosphine (28.8 mg, 0.11 mmol), and ditriphenylphosphine palladium dichloride (34.6 mg, 0.055 mmol) were added in sequence at room temperature (20°C). The reaction system was subjected to microwave reaction at 80°C for 2 hours under a nitrogen atmosphere. After the reaction of the raw material was completed as monitored by TLC, saturated aqueous ammonium chloride solution (50 mL) was added to the reaction solution, and extracted with ethyl acetate (40 mL×3). After the organic phases were combined, they were washed with saturated brine (2×20 mL), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dried under reduced pressure. The residue was purified by HPLC (Waters Sunfire OBD 100x30 mm, 5μm, mobile phase A: 0.1% TFA in water, mobile phase B: acetonitrile, gradient: 10% acetonitrile for 1 min, 52%-52% acetonitrile for 10 min, 95% acetonitrile for 14 min, and 10% acetonitrile for 16 min) to obtain compound 2F (30 mg, white solid, yield 3.9%).

[0199] MS (ESI): m / z: 355.6 [1 / 2M+H] + .

[0200] Step 6: Synthesis of Compound 2G

[0201] Compound 2F (30 mg, 0.042 mmol) was dissolved in ethanol (0.5 mL), and then 8% hydrochloric acid / ethyl acetate solution (1 mL) was added, and the mixture was heated to 40°C for 2 hours. After TLC showed that the reaction was complete, the reaction solution was directly spin-dried to obtain crude compound 2G, which was directly used in the next step (30 mg, yellow solid, 100.0%).

[0202] MS (ESI): m / z 610.1 [M+1] + .

[0203] Step 7: Synthesis of Compound 2

[0204] The intermediate INT-2 (27 mg, 0.071 mmol) was dissolved in N, N-dimethylformamide (0.5 mL), and N, N-diisopropylethylamine (30.3 mg, 0.24 mmol) and 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (29.8 mg, 0.078 mmol) were added to the reaction solution in sequence. After the addition, the mixture was reacted at 25°C for 15 minutes. Then, compound 2G (30 mg, 0.047 mmol) was dissolved in N, N-dimethylformamide (0.5 mL) and added dropwise to the reaction solution. After the addition, the mixture was stirred and reacted at 25°C for 1 hour. After TLC showed that the reaction was complete, the reaction solution was directly purified by HPLC (Waters Sunfire OBD 100x30 mm, 5μm, mobile phase A: 0.1% TFA in water, mobile phase B: acetonitrile, gradient: 10% acetonitrile for 1 min, 52%-52% acetonitrile for 10 min, 95% acetonitrile for 14 min, and 10% acetonitrile for 16 min) to obtain compound 2 (12 mg, off-white solid, yield 29.5%).

[0205] MS (ESI): m / z 481.7 [1 / 2M+1] + .

[0206] 1 H NMR(400MHz,dmso)δ12.83(s,1H),10.87(s,1H),8.86-8.76(m,2H),8.38(s,1H),8.18(dd,J=8.1,2.0Hz,1H),8.08(d,J=8.1Hz,1H),7.82(d, J=2.4Hz,2H),7.67(s,1H),7.50(d,J=3.6Hz,1H),7.48(d,J=8.1Hz,1H),7.43(s,1H),7.28(d,J=3.6Hz,1H),7.15(d,J=8.1Hz,1H),7.04(d,J =10.8Hz,1H),6.73(s,1H),4.37(d,J=12.4Hz,1H),4.22-3.93(m,5H),3.18-3.05(m,2H),2.98(br.s,2H),2.83(t,J=8.7Hz,1H),2.76-2.61( m,3H),2.60-2.56(m,1H),2.55-2.50(m,2H),2.46-2.42(m,1H),2.37- 2.24(m,2H),2.23-2.01(m,3H),1.88-1.64(m,6H),1.57-1.46(m,1H).

[0207] Example 3: Preparation of Compound 3

[0208] The structural formula of compound 3 is:

[0209]

[0210] The synthetic route of compound 3 is:

[0211]

[0212]

[0213] The specific preparation method of compound 3 includes:

[0214] Preparation method:

[0215] Step 1: Synthesis of compound 3B

[0216] Compound 3A (3 g, 16.13 mmol) and p-bromoiodobenzene (10 g, 35.48 mmol) were dissolved in toluene (82 mL). Sodium tert-butoxide (3.41 g, 35.48 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (1.57 g, 1.61 mmol) were added under stirring at room temperature. The reaction solution was replaced with nitrogen three times and stirred at 65°C for 16 hours. After TLC showed completion, the reaction solution was poured into water (200 mL) and extracted with dichloromethane (100 mL×3). The combined organic phase was washed with saturated brine (80 mL×2), dried over anhydrous sodium sulfate, filtered to remove the desiccant, desolventized under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 5:1 (volume ratio)) to obtain compound 3B (1.64 g, yellow solid, yield 29.9%).

[0217] MS (ESI): m / z 341.08 [M+1] + .

[0218] Step 2: Synthesis of Compound 3C

[0219] Compound 3B (1 g, 2.94 mmol) was dissolved in 1,4-dioxane (10 mL), and then diboric acid pinacol ester (1.49 g, 5.88 mmol), potassium acetate (0.86 g, 8.82 mmol), [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (0.29 g, 0.29 mmol) were added in sequence under stirring at room temperature. After the addition was complete, the reaction solution was replaced with nitrogen three times, and then stirred at 80°C for 4 hours. After TLC showed that the reaction was completed, the reaction solution was diluted with water (50 mL) and then extracted with ethyl acetate (40 mL×3). The combined organic phase was washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and desolventized under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5:1 (volume ratio)) to give compound 3C (1.01 g, white solid, yield 88.5%).

[0220] MS (ESI): m / z 389.25 [M+1] + .

[0221] Step 3: Synthesis of Compound 3D

[0222] Compound 3C (551.0 mg, 1.42 mmol) and intermediate INT-1 (500.0 mg, 1.09 mmol) were added to a mixed solvent of 1,4-dioxane (6 mL) and water (1 mL), and sodium carbonate (231.1 mg, 2.18 mmol), 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (93.4 mg, 0.22 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (94.3 mg, 0.13 mmol) were added in sequence at room temperature (20°C). The reaction system was subjected to microwave reaction at 80°C for 2 hours under a nitrogen atmosphere. After the reaction of the raw material was completed as monitored by TLC, saturated aqueous ammonium chloride solution (50 mL) was added to the reaction solution, and extracted with ethyl acetate (40 mL×3). After the organic phases were combined, they were washed with saturated brine (2×20 mL), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dried under reduced pressure. The residue was purified by HPLC (Waters Sunfire OBD 100x30 mm, 5μm, mobile phase A: 0.1% TFA in water, mobile phase B: acetonitrile, gradient: 10% acetonitrile for 1 min, 52%-52% acetonitrile for 10 min, 95% acetonitrile for 14 min, and 10% acetonitrile for 16 min) to obtain compound 3D (30 mg, white solid, yield 4.3%).

[0223] MS (ESI): m / z 322.1 [1 / 2M+1] + .

[0224] Step 4: Synthesis of compound 3E

[0225] Compound 3D (30 mg, 0.047 mmol) was dissolved in ethanol (0.5 mL), and then 8% hydrochloric acid / ethyl acetate solution (1 mL) was added, and the mixture was heated to 40°C for 2 hours. After TLC showed that the reaction was complete, the reaction solution was directly spin-dried to obtain crude compound 3E, which was directly used in the next step (30 mg, yellow solid, 100.0%).

[0226] MS (ESI): m / z 543.2 [M+1] + .

[0227] Step 5: Synthesis of Compound 3

[0228] Compound INT-2 (27 mg, 0.071 mmol) was dissolved in N,N-dimethylformamide (0.5 mL), and N,N-diisopropylethylamine (30.3 mg, 0.24 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (29.8 mg, 0.078 mmol) were added to the reaction solution in sequence. After the addition, the mixture was reacted at 25°C for 15 minutes. Then, 3E (30 mg, 0.047 mmol) was dissolved in N,N-dimethylformamide (0.5 mL) and added dropwise to the reaction solution. After the addition, the mixture was stirred at 25°C for 1 hour. After TLC showed that the reaction was complete, the reaction solution was directly purified by HPLC (Waters Sunfire OBD 100x30 mm, 5μm, mobile phase A: 0.1% TFA in water, mobile phase B: acetonitrile, gradient: 10% acetonitrile for 1 min, 52%-52% acetonitrile for 10 min, 95% acetonitrile for 14 min, and 10% acetonitrile for 16 min) to obtain compound 3 (14 mg, off-white solid, yield 34.0%).

[0229] MS (ESI): m / z 448.2 [1 / 2M+1] + .

[0230] 1H NMR(400MHz,dmso)δ12.79(s,1H),10.88(s,1H),9.52(s,1H),8.25(s,1H),7.86(s,1H),7.66(dd,J=15.8,6.8Hz,3H),7.53 (d,J=8.1Hz,1H),7.50(d,J=3.6Hz,1H),7.44(s,1H),7.27(d,J=3.6Hz,1H),7.19-7.11(m,2H),7.07(d,J=8.6Hz,2H),6.69( s,1H),4.44-4.30(m,2H),4.10(dd,J=11.7,4.5Hz,1H),4.06-3.96(m,2H),3.73-3.65(m,2H),3.63-3.50(m,3H),3.29-3.1 9(m,4H),3.17-3.06(m,2H),3.00-2.89(m,2H),2.85-2.68(m,3H),2.61-2.50(m,3H),2.31-2.24(m,1H),2.16-1.81(m,6H).

[0231] Experimental Example 1: Cell Antiproliferation Activity Experiment

[0232] (1) Experimental materials:

[0233] RPMI1640, glutamine, interleukin-3, and trypsin were purchased from Life Technology.

[0234] Fetal bovine serum and double-antibody were purchased from Hyclone.

[0235] Blasticidin was purchased from Merck.

[0236] Phosphate buffered saline was purchased from Corning.

[0237] CellTiter-Glo kit was purchased from Promega.

[0238] Ba / F3 cells were purchased from Riken.

[0239] Ba / F3 (EGFR L858R / T790M / C797S) cells were constructed by Zhongke Pusheng.

[0240] Ba / F3 cell culture medium: 88% RPMI 1640, 10% fetal bovine serum, 10 ng / mL interleukin 3, 1% glutamine, 1% double antibody.

[0241] Ba / F3 (EGFR L858R / T790M / C797S) cell culture medium: 87.9% RPMI 1640, 10% fetal bovine serum, 0.1% blasticidin, 1% glutamine, and 1% double antibody.

[0242] Experimental board: 781091 (Greiner)

[0243] Plate reader: Envision (PerkinElmer)

[0244] (2) Experimental methods:

[0245] The test compound was diluted 3-fold with DMSO at 10 points on ECHO, and 250 nL of the compound was transferred to a 384-well plate, with a final concentration of 5-0.00025 μM. 250 nL of DMSO was transferred to the positive control and negative control. The cells in the T75 culture flask were evenly pipetted and counted. The concentration of Ba / F3 (EGFR L858R / T790M / C797S) cells was adjusted to 4×10 4 100 μL of phosphate buffer was added to the outer wells of the 384-well plate, and 50 μL of cell suspension was added to the other wells. After standing at room temperature for 15 min, it was placed at 37°C with 5% CO 2 Incubate in an incubator for 72 h; add 25 μL CellTiter-Glo to each well of a 384-well plate, centrifuge at 1000 rpm for 15 s, shake at room temperature for 15 min, and read using Envision.

[0246] (3) Data analysis:

[0247] The readings were converted into inhibition rate (%) by the following formula: (Sample value-HC) / (LC-HC)*100. IC was determined by parameter curve fitting (XL-fit software). 50 data.

[0248] (4) Test results:

[0249] The Ba / F3 (EGFR L858R / T790M / C797S) cell activity inhibitory activity data of the compounds of the present invention are shown in the following Table 1.

[0250] Table 1 provides the inhibitory activity of the compounds of the present invention on the proliferation of Ba / F3 (EGFR L858R / T790M / C797S) cells.

[0251] Table 1 Anti-cell proliferation activity data of the compounds of the present invention (IC 50 )

[0252]

[0253] From the experimental results in Table 1, it can be seen that the compounds of the present invention have good activity in inhibiting the proliferation of Ba / F3 (EGFR L858R / T790M / C797S) cells. The activity of many compounds is less than 100nM. It shows extremely important anti-tumor potential and has good clinical application prospects.

[0254] Experimental Example 2: Pharmacokinetics Experiment

[0255] 1. Experimental Materials

[0256] The compound prepared in the above example was used to prepare a 4.76 mg / mL clear solution (5% DMSO + 30% PEG300 + 2% Tween80 + 63% H 2 O).

[0257] 2. Experimental Animals

[0258] Male ICR mice, SPF grade, 3 in total, weighing 27-28 g, were provided by Shanghai Slake Laboratory Animal Co., Ltd. The test mice were given a 2-4 day environmental adaptation period before the experiment, and the animals were fed normally before drug administration.

[0259] 3. Experimental Methods

[0260] 1) After the mice had eaten normally, blank plasma was collected at time 0;

[0261] 2) taking the mice prepared in step 1), and orally (PO) administering 50 mg / kg of the test compound;

[0262] 3) Blood samples were collected from the cheek at 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h, and 24h after oral administration. About 0.05mL of blood was collected from each sample, anticoagulated with sodium heparin, and placed on wet ice after collection. Blood samples were placed on ice after collection and centrifuged within 1 hour to separate plasma (centrifugation conditions: 6000g, 3 minutes, 2-8℃). Plasma samples were stored in a -80℃ refrigerator before analysis.

[0263] 4) According to step 3), the pharmacokinetic parameters were calculated using Phoenix WinNonlin8.2.0 using the blood drug concentration data at different time points, providing parameters such as AUC0-t, AUC0-∞, MRT0-∞, Cmax, Tmax, and T1 / 2 and their mean values ​​and standard deviations. The test results are shown in Table 2 below.

[0264] Table 2 Pharmacokinetic data of the compounds of the present invention

[0265]

[0266] As shown in Table 2, after oral or intravenous administration to mice or rats, the compounds of the present invention have a longer half-life and a higher exposure in the animal plasma, and can be administered orally.

[0267] Although the embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and cannot be understood as limiting the present invention. Without departing from the principles and purpose of the present invention, those skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention, and these changes, modifications, replacements and modifications are all within the scope of the present invention.

Claims

1. Compounds that inhibit / induce degradation of EGFR kinase, It is characterized in that The compound is a compound as shown in any one of Formulae 1 to 3 or a pharmaceutically acceptable salt or racemate thereof, 。 2. Pharmaceutical composition, It is characterized in that The invention comprises an effective dose of one or more of the compound according to claim 1, its pharmaceutically acceptable salt, and its racemate.

3. The pharmaceutical composition according to claim 2, It is characterized in that The pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient.

4. The pharmaceutical composition according to claim 3, It is characterized in that The auxiliary materials include pharmaceutically acceptable excipients or therapeutic agents.

5. Use of one or more of the compound according to claim 1, its pharmaceutically acceptable salt, and its racemate in the preparation of a medicament for treating and / or preventing EGFR kinase-mediated diseases.

6. Use of one or more of the compound according to claim 1, its pharmaceutically acceptable salt, and its racemate in the preparation of drugs for treating and / or preventing cancer.

7. The use according to claim 6, It is characterized in that The cancer includes ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, renal cancer, multiple myeloma or mesothelioma.

Images