Heterocyclic compounds and uses thereof

Abstract

The application discloses a kind of and heterocyclic compound and its use, belong to chemical medicine technical field.The and heterocyclic compound shown in formula II provided by the application can be used as PARP1 inhibitor, with high activity high selectivity advantage, also has excellent pharmacokinetic property and excellent safety.

Description

Technical Field

[0001] This invention belongs to the field of chemical medicine and relates to a class of heterocyclic compounds and their uses. Background Technology

[0002] During cell growth, its DNA is constantly damaged by various internal and external adverse factors. Among DNA damage types, the most serious are single-strand breaks and double-strand breaks, with single-strand breaks being more common. If these breaks are not repaired promptly and accurately, they can lead to genomic instability, subsequently causing carcinogenesis or even cell death. For single-strand DNA breaks, repair primarily relies on PARP enzymes. For double-strand breaks, repair mechanisms include non-homologous end linkage and homologous recombination repair. Homologous recombination repair is a high-fidelity, error-free repair method and the main pathway for double-strand DNA repair. Many proteins are involved in homologous recombination repair, the most well-known being BRCA proteins. Two studies in 2005 (Farmer H, McCabe N, et al. Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy[J]. Nature, 2005, 434(7035):917-921. Bryant, H., Schultz, N., Thomas, H. et al. Specific killing of BRCA2-deficient tumors with inhibitors of poly(ADP-ribose)polymerase. Nature 434, 913–917(2005)) showed that tumor cells lacking BRCA1 or BRCA2 were selectively inhibited by PARP inhibitors. Based on this research, researchers proposed the concept of synthetic lethality: the deletion of either BRCA or PARP genes is not fatal in itself, but the simultaneous inactivation of both leads to cell death. Based on the theory of synthetic lethality, PARP inhibitors (PARPi) have been developed to selectively target cancer cells with BRCA1 / 2 mutations.

[0003] PARP inhibitors have demonstrated excellent clinical efficacy in patients with homologous recombination-deficient cancers. However, whether used as monotherapy or in combination therapy, hematological toxicities (anemia, neutropenia, and thrombocytopenia) and other toxicities limit the application of these drugs. Related research (Harris PA, Boloor A, Cheung M, et al. Discovery of 5-[[4-[(2,3-dimethyl-2H-indazol-6-yl)methylamino]-2-pyrimidinyl]amino]-2-methyl-benzenesulfon amide (Pazopanib), a novel and potent vascular endothelial growth factor receptor inhibitor.[J]. Journal of Medicinal Chemistry, 2008, 51(15):4632.) suggests that these adverse reactions may stem from the inhibition of PARP2 by marketed PARP inhibitors, which is not essential for therapeutic efficacy. Highly selective PARP1 inhibitors can reduce hematological toxicity, improve the therapeutic safety window, and increase the potential for combination therapy with other chemotherapy or targeted drugs.

[0004] Therefore, there is an unmet clinical need for effective and safe PARP inhibitors, particularly PARP inhibitors selective for PARP1. The novel PARP1 inhibitor described in this invention exhibits unexpectedly high selectivity for PARP1 compared to other PARP family members (such as PARP2, PARP3, PARP5a, and PARP6), and can be used to treat diseases related to PARP function. Summary of the Invention

[0005] The purpose of this invention is to provide a class of heterocyclic compounds and their uses to achieve highly selective and efficient prevention or treatment of diseases related to PARP function.

[0006] In a first aspect, the present invention provides a compound of Formula II or a pharmaceutically acceptable form thereof, said Formula II having the following structure:

[0007]

[0008] in:

[0009] Indicates a single or double bond;

[0010] R1 is selected from C 1-4 Deuterated alkyl groups;

[0011] X1 is selected from N or C(R)5a X2 is selected from N or C(R). 5b X3 is selected from N or C(R) 5c X1, X2, and X3 have exactly one selected from N;

[0012] R 2a and R 2b It is independently selected from hydrogen, deuterium, methyl, or deuterated methyl;

[0013] R3 is selected from deuterium, fluorine, and C. 1-4 Alkyl or C 1-4 At least one of the deuterated alkyl groups;

[0014] R 3a Selected from hydrogen, deuterium, fluorine, hydroxyl, cyano, C 1-4 Alkyl, C 1-4 Fluorinated alkyl or C 1-4 Alkoxy;

[0015] R4 is selected from hydrogen, halogen, cyano, and C. 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluorinated alkyl groups or -CONHR7;

[0016] R 5a R 5a and R 5c Independently selected from hydrogen, fluorine, chlorine, and C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluoroalkyl, C 1-4 Alkoxy or C 1-4 Fluoroalkoxy;

[0017] R6 is selected from hydrogen, fluorine, chlorine, and C. 1-4 Alkyl, C 1-4 Fluorinated alkyl or C 1-4 Deuterated alkyl groups;

[0018] R7 is selected from hydrogen, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluorinated alkyl groups or 3-6 membered cycloalkyl groups;

[0019] X5 is selected from nitrogen or C(R) 9a X6 is selected from nitrogen or C(R) 9b X7 is selected from nitrogen or C(R) 9c X8 is selected from nitrogen or C(R) 9d );

[0020] R 9a R 9b R 9c and R 9dIndependently selected from hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluoroalkyl, C 1-4 Alkoxy or C 1-4 Fluoroalkoxy;

[0021] n1 is an integer between 0 and 8;

[0022] n3 is independently selected from 0 or 1;

[0023] The pharmaceutically acceptable form is selected from pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled substances, metabolites, or prodrugs.

[0024] In some embodiments of the present invention, in the compound represented by Formula II above or in its pharmaceutically acceptable form, R1 is selected from deuterated methyl or deuterated ethyl.

[0025] In some preferred embodiments of the present invention, in the compound represented by Formula II above or in its pharmaceutically acceptable form, R1 is selected from -CD2CD3 or -CH2CD3.

[0026] In some embodiments of the present invention, in the compound represented by Formula II above or in its pharmaceutically acceptable form, R3 is selected from at least one of deuterium, methyl, deuterated methyl, fluoromethyl or methoxy.

[0027] In some preferred embodiments of the present invention, in the compound represented by Formula II above or in its pharmaceutically acceptable form, R3 is selected from deuterium or methyl.

[0028] In some embodiments of the present invention, the compound represented by Formula II above, or its pharmaceutically acceptable form, is used. When it is a double bond, R 3a It does not exist. When it is a single bond, R 3a Selected from hydrogen and deuterium.

[0029] In some embodiments of the present invention, in the compound represented by Formula II above, or in its pharmaceutically acceptable form, R4 is selected from fluorine, chlorine, cyano, C... 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluoroalkyl, C 1-4 Alkyl, C 1-4 Deuterated alkylamino acyl, C 1-4 Fluoroalkylaminoacyl or 3-6 membered cycloalkylaminoacyl.

[0030] In some preferred embodiments of the present invention, in the compound represented by Formula II above or in its pharmaceutically acceptable form, R4 is selected from fluorine, cyano, methylaminoacyl, deuterated methylaminoacyl or cyclopropylaminoacyl.

[0031] In some preferred embodiments of the invention, in the compound represented by Formula II above or in its pharmaceutically acceptable form, R6 is selected from hydrogen, fluorine, chlorine or methyl.

[0032] In some embodiments of the present invention, in the compounds represented by Formula II above or in their pharmaceutically acceptable form, R 5a R 5b and R 5c It is independently selected from hydrogen, fluorine, chlorine, or methyl.

[0033] In some embodiments of the present invention, based on the compound represented by Formula II above or its pharmaceutically acceptable form, the present invention also provides compounds having the structure described in Formula VI-1:

[0034]

[0035] in:

[0036] R6 is selected from hydrogen or fluorine;

[0037] R 9a Selected from hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Fluorinated alkyl groups;

[0038] n1 is an integer between 0 and 6.

[0039] In some embodiments of the present invention, based on the compound represented by Formula II above or its pharmaceutically acceptable form, the present invention also provides compounds having the structure described in Formula VI-3:

[0040]

[0041] in:

[0042] R6 is selected from hydrogen or fluorine;

[0043] R 9a Selected from hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Fluorinated alkyl groups;

[0044] n1 is an integer between 0 and 6.

[0045] In some more preferred embodiments of the present invention, the following compounds and their pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled substances, metabolites, or prodrugs are provided:

[0046]

[0047] In some more preferred embodiments of the present invention, the following compounds and their pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled substances, metabolites, or prodrugs are provided:

[0048]

[0049] In some more preferred embodiments of the present invention, the following compounds and their pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled substances, metabolites, or prodrugs are provided:

[0050]

[0051] In a second aspect, the present invention provides a pharmaceutical composition having the aforementioned compound (Formula II, Formula VI-1 or Formula VI-3) or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite or prodrug as the active ingredient, supplemented by a pharmaceutically acceptable carrier.

[0052] A further object of the present invention is to provide a method for preparing the pharmaceutical composition of the present invention, the method comprising combining any compound of formula II, formula VI-1 or formula VI-3 or a pharmaceutically acceptable form thereof, or a mixture thereof, with one or more pharmaceutically acceptable carriers.

[0053] The pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions of the present invention are pharmaceutically acceptable carriers, and examples of suitable pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (2005).

[0054] Pharmaceutical compositions can be administered in any form, as long as they achieve the purpose of preventing, alleviating, preventing, or curing symptoms in human or animal patients. For example, they can be formulated into various suitable dosage forms depending on the route of administration.

[0055] In other embodiments, the administration of the compounds or pharmaceutical compositions of the present invention may be combined with other treatment methods. These other treatment methods may be selected from, but are not limited to, radiotherapy, chemotherapy, immunotherapy, or combinations thereof.

[0056] This invention also relates to a pharmaceutical formulation comprising any compound of formula II, formula VI-1 or formula VI-3 or a pharmaceutically acceptable form thereof, or a mixture thereof, as an active ingredient, or the pharmaceutical composition of this invention. In some embodiments, the formulation is in the form of a solid dosage form, a semi-solid dosage form, a liquid dosage form, or a gaseous dosage form.

[0057] A further object of the present invention is to provide an article of manufacture, for example, in the form of a kit. The article of manufacture as used herein is intended to include, but is not limited to, medicine boxes and packaging. The article of manufacture of the present invention comprises: (a) a first container; (b) a pharmaceutical composition contained in the first container, wherein the composition comprises: a first therapeutic agent comprising: any compound of formula II, formula VI-1 or formula VI-3 or a pharmaceutically acceptable form thereof, or a mixture thereof; (c) optionally present packaging instructions stating that the pharmaceutical composition may be used to treat oncological conditions (as defined below); and (d) a second container.

[0058] The first container is a container for containing a pharmaceutical composition. This container may be used for the preparation, storage, transportation, and / or individual / bulk sales. The first container is intended to encompass bottles, jars, vials, flasks, syringes, tubes (e.g. for cream products), or any other container for the preparation, containment, storage, or dispensing of pharmaceutical products.

[0059] The second container is a container for holding the first container and optional instruction manuals. Examples of the second container include, but are not limited to, boxes (e.g., cardboard or plastic boxes), cartons, cartons, bags (e.g., paper or plastic bags), sachets, and cloth bags. The instruction manuals may be physically attached to the outside of the first container by cable ties, glue, U-staples, or other adhesive methods, or they may be placed inside the second container without any physical means of attachment to the first container. Alternatively, the instruction manuals may be located outside the second container. When located outside the second container, it is preferable that the instruction manuals be physically attached by cable ties, glue, U-staples, or other adhesive methods. Alternatively, they may be adjacent to or in contact with the outside of the second container without physical attachment.

[0060] The package insert, such as a trademark, label, or marking, lists information relating to the pharmaceutical composition contained within the first container. The listed information is typically determined by the regulatory authority governing the region where the product is to be sold (e.g., the U.S. Food and Drug Administration). Preferably, the package insert specifically lists the approved indications for which the pharmaceutical composition is used. The package insert can be made of any material from which information contained therein or on the material can be read. Preferably, the package insert is made of a printable material (e.g., paper, plastic, cardboard, foil, adhesive paper, or plastic) on which the desired information can be formed (e.g., printed or coated).

[0061] Thirdly, the present invention provides the use of the aforementioned compounds, compounds of formula II, formula VI-1 or formula VI-3, and related specific compounds or their pharmaceutically acceptable forms, or the use of the pharmaceutical compositions of the present invention in the preparation of medicaments for the prevention or treatment of PARP1 enzyme-related diseases.

[0062] The present invention provides a method for the prevention or treatment of PARP1 enzyme-related diseases, the method comprising administering to an individual in need a compound of formula II, formula VI-1 or formula VI-3 or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention.

[0063] The present invention provides compounds of formula II, formula VI-1 or formula VI-3 or their pharmaceutically acceptable forms, or pharmaceutical compositions of the present invention, for the prevention or treatment of PARP1 enzyme-related diseases.

[0064] The present invention provides a method for preventing or treating PARP1 enzyme-related diseases by combining a compound of formula II, formula VI-1 or formula VI-3 or a pharmaceutically acceptable form thereof, or a pharmaceutical composition thereof, in combination with other treatment methods, including but not limited to: radiotherapy, chemotherapy, immunotherapy, or combinations thereof.

[0065] In some implementations, the PARP1 enzyme-related disease is a disease that is sensitive to or responsive to PARP1 enzyme inhibition.

[0066] In some implementations, the PARP1 enzyme-related disease is a tumor-related condition.

[0067] In some preferred embodiments, the tumor-related diseases lack the HR-dependent DNA DSB repair pathway.

[0068] In some preferred embodiments, the tumor-like condition comprises one or more cancer cells that have a reduced or absent ability to repair DNA DSB via HR compared to normal cells.

[0069] In some preferred embodiments, the cancer cells have a BRCA1 or BRCA2 defective phenotype.

[0070] In some embodiments, the PARP1 enzyme-related diseases are oncological conditions, including but not limited to solid and hematologic malignancies. In further embodiments, the oncological conditions include, but are not limited to, breast cancer, colorectal cancer, colon cancer, lung cancer (including small cell lung cancer, non-small cell lung cancer, and bronchioloalveolar carcinoma), and prostate cancer, as well as bile duct cancer, bone cancer, bladder cancer, head and neck cancer, kidney cancer, liver cancer, gastrointestinal cancer, esophageal cancer, ovarian cancer, pancreatic cancer, skin cancer, testicular cancer, thyroid cancer, uterine cancer, cervical cancer, and vulvar cancer, as well as leukemia (including chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), and chronic myeloid leukemia (CML)), multiple myeloma, or lymphoma.

[0071] In some preferred embodiments, the PARP1 enzyme-related diseases are breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer, or lung cancer.

[0072] In a further preferred embodiment, the compounds of the present invention can be used in combination with radiotherapy, chemotherapy, or immunotherapy to prevent or treat cancer.

[0073] The beneficial effects of this invention are:

[0074] This invention provides a novel class of highly active and selective PARP1 inhibitors that can achieve at least one of the following technical effects: (1) high inhibitory activity against PARP1 enzyme; (2) selective inhibition of PARP1 enzyme, with high selectivity for other PARP family enzymes such as PARP2, PARP5a, and PARP5b; (3) strong inhibitory activity against homologous recombination-deficient tumor cells, with weak inhibitory effect on non-homologous recombination-deficient cells; (4) excellent pharmacokinetic properties (e.g., good bioavailability, suitable half-life, and duration of action); and (5) excellent safety (low toxicity and / or fewer side effects, and a wider therapeutic window).

[0075] Terminology definition:

[0076] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by those skilled in the art. The terms “comprising,” “including,” “having,” “containing,” or “involving,” and their other variations herein, are inclusive or open-ended and do not exclude other unlisted elements or method steps. Those skilled in the art will understand that the foregoing term “comprising” encompasses the meaning of “consisting of.”

[0077] In this invention, the terms "a," "an," "the," "at least one," and "one or more" are used interchangeably. Thus, for example, a composition comprising "a" pharmaceutically acceptable excipient can be interpreted as indicating that the composition comprises "one or more" pharmaceutically acceptable excipients.

[0078] For example, the expression "C" 1-4 "This should be understood as encompassing any subrange and each point value, such as C." 2-4 C 3-4 C 1-2 C 1-3 C 1-4 And so on, as well as C1, C2, C3, C4, etc.

[0079] Unless otherwise stated, all other uses of this material are permitted. It indicates a single or double bond.

[0080] In this invention, unless otherwise stated, halogen refers to fluorine, chlorine, bromine or iodine.

[0081] In this invention, unless otherwise stated, "alkyl" includes straight-chain or branched monovalent saturated hydrocarbon groups. For example, alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 3-(2-methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl, 2-methylpentyl, etc. Similarly, "C 1-4 C in "alkyl" 1-4 It refers to a group that contains 1, 2, 3 or 4 carbon atoms arranged in a straight chain or branched chain.

[0082] In this invention, unless otherwise stated, “cycloalkyl,” “carbocyclic,” or “cycloalkylene” refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic) non-aromatic hydrocarbon group. Common cycloalkyl groups include (but are not limited to) monocyclic cycloalkyl groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclobutene, cyclopentene, cyclohexene, etc.; or bicyclic cycloalkyl groups, including fused rings, bridged rings, or spirocyclic groups, such as bicyclic [1.1.1]pentyl, bicyclic [2.2.1]heptyl, bicyclic [3.2.1]octyl, bicyclic [5.2.0]nonyl, decahydronaphthyl, etc. For example, “C 3-12 cycloalkyl” refers to a cycloalkyl group having 3-12 cyclic carbon atoms (such as 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12). The cycloalkyl or cycloalkylene groups in this invention may optionally be substituted by one or more substituents described in this invention.

[0083] In this invention, unless otherwise stated, "fluoroalkyl" refers to the alkyl group described above, wherein one or more hydrogen atoms are replaced by fluorine atoms. For example, the term "C 1-4"Fluoroalkyl" refers to a C-aryl group that is optionally substituted with one or more (e.g., 1-3) fluorine atoms. 1-4 Alkyl groups. Those skilled in the art will understand that when there is more than one fluorine atom substituent, the fluorine atoms can be the same or different, and can be located on the same or different C atoms. Examples of haloalkyl groups include, for example, -CH2F, -CHF2, -CF3, -C2F5, -CH2CF3, -CH2CH2CF3, etc. The fluoroalkyl groups in this invention are optionally substituted with one or more substituents described in this invention.

[0084] This invention also includes all pharmaceutically acceptable isotopically labeled compounds that are identical to the compounds of this invention, except that one or more atoms are replaced by atoms having the same atomic number but with an atomic mass or mass number different from the dominant atomic mass or mass number in nature. Examples of isotopes suitable for inclusion in the compounds of this invention include (but are not limited to) isotopes of hydrogen (e.g., deuterium). 2 H), tritium ( 3 H); carbon isotopes (e.g., ... 13 C and 14 C); isotopes of chlorine (e.g., 37Cl); isotopes of iodine (e.g., 125 I); nitrogen isotopes (e.g.) 13 N and 15 N); isotopes of oxygen (e.g., N); 17 O and 18 O); isotopes of phosphorus (e.g., O); phosphorus isotopes (e.g., O); 32 P); and isotopes of sulfur (e.g., ... 34 S).

[0085] In this invention, "polymorph" refers to different solid crystalline phases resulting from the presence of two or more different molecular arrangements in the solid state of certain compounds of this invention. Some compounds of this invention may exist in more than one crystal form, and this invention aims to include various crystal forms and mixtures thereof. Typically, crystallization produces solvates of the compounds of this invention. The term "solvate" as used in this invention refers to an aggregate comprising one or more molecules of the compound of this invention and one or more solvent molecules. The solvent may be water, in which case the solvate is a hydrate. Alternatively, the solvent may be an organic solvent. Therefore, the compounds of this invention can exist as hydrates, including monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, etc., and corresponding solvated forms. The compounds of this invention can form true solvates, but in some cases, they may also remain only as indeterminate water or a mixture of water and a portion of indeterminate solvent. The compounds of this invention can react in a solvent or precipitate or crystallize from a solvent. The solvates of the compounds of this invention are also included within the scope of this invention. This invention also covers all possible crystalline forms or polymorphs of the compounds of this invention, which may be a single polymorph or a mixture of more than one polymorph in any proportion.

[0086] In this invention, "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In compounds having one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, diastereomer mixtures, and individual diastereomers can be produced. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of this invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, and imine-enamine tautomers. It is to be understood that the scope of this invention covers all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0087] In this invention, pharmaceutically acceptable salts include their acid addition salts and base addition salts. Suitable acid addition salts are formed by acids that form pharmaceutically acceptable salts. Suitable base addition salts are formed by bases that form pharmaceutically acceptable salts. A review of suitable salts can be found, for example, in “Remington’s Pharmaceutical Sciences,” Mack Publishing Company, Easton, Pa., (2005); and “Handbook of Pharmaceutical Salts: Properties, Selection, and Use,” Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of this invention are known to those skilled in the art. “Pharmaceutically acceptable acid addition salt” refers to a salt formed with an inorganic or organic acid that retains the bioavailability of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobroms, sulfates, nitrates, and phosphates; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetate, trifluoroacetate, propionate, hexanoate, octanoate, decanoate, undecanoate, glycolate, gluconate, lactate, sebate, adipate, glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate, cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, alginate, ascorbate, salicylate, 4-aminosalicylic acid, and naphthalenedisulfonate. These salts can be prepared by methods known in this patent. "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that retain the bioavailability of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, and aluminum salts. Preferred inorganic salts are ammonium salts, sodium salts, calcium salts, and magnesium salts. Salts derived from organic bases include, but are not limited to, the following: primary amines, secondary amines, and tertiary amines; substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, and polyamine resins.Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in this patent.

[0088] In this invention, unless otherwise stated, "ester" refers to an ester derived from the compounds described herein, including physiologically hydrolyzable esters (compounds of this invention that can be hydrolyzed under physiological conditions to release free acids or alcohols). The compounds of this invention may themselves be esters.

[0089] The compounds of the present invention can exist as solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, can be stoichiometric or non-stoichiometric.

[0090] Those skilled in the art will understand that not all nitrogen-containing heterocycles can form nitrogen oxides because nitrogen requires available lone pairs of electrons to be oxidized. Those skilled in the art will identify nitrogen-containing heterocycles capable of forming nitrogen oxides. They will also recognize that tertiary amines can form nitrogen oxides. Synthetic methods for preparing nitrogen oxides of heterocycles and tertiary amines are well known to those skilled in the art, including the oxidation of heterocycles and tertiary amines with peroxyacids such as peracetic acid and m-chloroperoxybenzoic acid (mCPBA), hydrogen peroxide, alkyl peroxides such as tert-butyl peroxide, sodium perborate, and dioxiranes such as dimethyldioxirane. These methods for preparing nitrogen oxides have been extensively described and reviewed in the literature, see, for example: T.L. Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp. 748-750 (AR. Katritzky and A.J. Boulton, Eds., Academic Press); and G.W. H. Heeseman and E.S. G. Wierstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp. 390-392 (AR. Katritzky and A.J. Boulton, Eds., Academic Press).

[0091] In this invention, "metabolite" refers to a substance formed in the body upon administration of a compound of the present invention. Metabolites of the compound can be identified using techniques known in the art, and their activity can be characterized by experimental methods. Such products can be generated, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc., of the administered compound. Therefore, this invention includes metabolites of the compounds of the present invention, including compounds obtained by methods that expose the compounds of the present invention to mammals for a time sufficient to produce their metabolites.

[0092] In this invention, a "prodrug" refers to certain derivatives of the compounds of the invention that, when administered to or onto the body, can be converted, for example, by hydrolysis and cleavage into the compounds of the invention having the desired activity. Typically, such prodrugs are functional group derivatives of the compounds that readily convert in vivo into the desired therapeutically active compounds. Further information regarding the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems," Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella). The prodrugs of the invention can be prepared, for example, by replacing suitable functional groups present in the compounds of the invention with portions known to those skilled in the art as "pro-moiety" (e.g., as described in "Design of Prodrugs," H. Bundgaard (Elsevier, 1985)).

[0093] In this application, "pharmaceutical composition" refers to a formulation of the compounds of the present invention with a medium generally accepted in the art for delivering bioactive compounds to mammals (e.g., humans). This medium includes pharmaceutically acceptable carriers. The purpose of the pharmaceutical composition is to facilitate administration to the organism, enhance the absorption of the active ingredient, and thereby exert its bioactivity.

[0094] In this application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, flow aid, sweetener, diluent, preservative, dye / coloring agent, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that is permitted by the relevant government regulatory authority or is acceptable for human or livestock use.

[0095] As used herein, the terms “drug combination,” “drug co-administration,” “combination therapy,” “administration of other treatments,” and “administration of other therapeutic agents” refer to pharmaceutical treatments achieved by mixing or combining more than one active ingredient, including fixed and non-fixed combinations of active ingredients. The term “fixed combination” refers to the simultaneous administration to a patient of at least one compound described herein and at least one synergistic agent in the form of a single entity or dosage form. The term “non-fixed combination” refers to the simultaneous, combined, or sequential administration to a patient of at least one compound described herein and at least one synergistic agent in the form of a single entity at variable intervals. These also apply to cocktail therapies, such as the administration of three or more active ingredients.

[0096] In this invention, unless otherwise stated, "tumor" includes, but is not limited to, diseases such as leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, squamous cell carcinoma of the lung, adenocarcinoma of the lung, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, cervical cancer, ovarian cancer, intestinal cancer, nasal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, and oral cancer.

[0097] In this invention, unless otherwise stated, "treatment" means reversing, alleviating, or inhibiting the progression of a disease or condition or one or more symptoms of such a disease or condition, or preventing such a disease or condition or one or more symptoms of such a disease or condition.

[0098] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention. Attached Figure Description

[0099] Figure 1 The figure shows the changes in tumor volume in the MDA-MB-436 nude mouse model after administration of compounds 1 and 6.

[0100] Figure 2 The tumor volume changes in the MDA-MB-436 nude mouse model after administration of compounds 1, 6 and 21 are shown in the figure.

[0101] Figure 3 This is a graph showing the changes in tumor volume in a PDX model mouse with gastric cancer after compound 6 was used in combination with carboplatin.

[0102] Figure 4 The tumor volume in the SUM14PT nude mouse subcutaneous xenograft model after compound 6 was combined with carboplatin. Detailed Implementation

[0103] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they shall be performed in accordance with the techniques or conditions described in the literature in the art or in accordance with the product manual.

[0104] The reagents and raw materials used in the embodiments of this invention are all commercially available.

[0105] Table 1. Abbreviations and their meanings in this invention.

[0106]

[0107] The structure of the compound described in this invention was determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS). Chemical shifts were expressed as 10⁻⁶. -6 (ppm) is given as the unit.

[0108] MS measurements were performed using an Agilent SQD (ESI) mass spectrometer (manufacturer: Agilent, signal: 6110).

[0109] HPLC determinations were performed using an Agilent 1200DAD high-performance liquid chromatograph (Sunfirc C18, 150X4.6mm, 5µm column) and a Waters 2695-2996 high-performance liquid chromatograph (Gimini C18, 150X4.5mm, 5µm column).

[0110] The silica gel plates used for thin-layer chromatography are Qingdao Ocean GF254 silica gel plates. The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15mm-0.2mm, while the silica gel plates used for thin-layer chromatography separation and purification of products have a diameter of 0.4mm-0.5mm.

[0111] Column chromatography typically uses Qingdao Marine 100-200 or 200-300 mesh silica gel as a carrier.

[0112] Unless otherwise specified in the following examples, all reactions are carried out under an argon or nitrogen atmosphere. An argon or nitrogen atmosphere refers to a reaction flask connected to an approximately 1L argon or nitrogen balloon. A hydrogen atmosphere refers to a reaction flask connected to an approximately 1L hydrogen balloon. Hydrogenation reactions are typically performed under vacuum, followed by hydrogen filling, and repeated three times.

[0113] intermediate preparation

[0114] Intermediate int-1: N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0115]

[0116] Step 1: Compounds int-1a (1 g, 4.6 mmol), int-1b (1.7 g, 5.5 mmol), Pd(dppf)Cl2 (0.3 g, 0.46 mmol), and potassium carbonate (1.6 g, 11.5 mmol) were added to a mixed solvent of 7 ml dioxane, 3 ml anhydrous ethanol, and 4 ml water. The mixture was then purged with nitrogen three times and reacted at 90 °C for 2 h under nitrogen protection. After TLC detection of complete reaction, the reaction mixture was cooled to room temperature, and 30 ml dichloromethane and 20 ml water were added. The mixture was separated into layers in a separatory funnel. The aqueous phase was extracted twice with dichloromethane. The combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography to obtain compound int-1c (1 g, white solid).

[0117] Step 2: Add int-1c (1g, 3mmol), methylamine aqueous solution (5g, 161.3mmol), and anhydrous methanol (20ml) to a 100ml reaction flask, and stir overnight at room temperature. After the reaction is complete as monitored by TLC, concentrate the reaction solution under reduced pressure to dryness to obtain compound int-1d (0.8g, white solid).

[0118] Step 3: Add compound int-1d (0.5g, 1.5mmol) to 10ml of anhydrous methanol, followed by 10ml of 4mol / L dioxane hydrochloride solution. Stir at room temperature for 0.5-1h. After the reaction is complete as monitored by TLC, concentrate the reaction solution under reduced pressure to obtain compound int-1 (0.5g, white solid).

[0119] Example 1: 1'-(7-ethyl-6-oxo-5,6-dihydro-1,5-naphthid-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0120]

[0121] Step 1: Compound 1a (20 g, 95.1 mmol) and selenium dioxide (16 g, 144 mmol) were added to a 250 mL reaction flask, followed by 120 mL of 1,4-dioxane. The mixture was heated to 110 °C and stirred overnight. After the reaction was complete as monitored by TLC, the reaction solution was filtered, and the residue was washed with ethyl acetate. The filtrates were combined and concentrated by rotary evaporation. The crude product was purified by column chromatography to give compound 1b (16 g, yellow solid). LC-MS: ESI [M+H]+ =225.2.

[0122] Step 2: Add sodium hydride (6.86 g, 171.4 mmol) to a 250 ml reaction flask, then add 60 ml of 1,4-dioxane, and purge with nitrogen three times. Cool to 0 °C, and slowly add triethyl 2-phosphonobutyrate (43.2 g, 171.4 mmol) under nitrogen protection. Stir the reaction at 0 °C for 10 minutes, then heat to room temperature and stir for 10 minutes, then heat to 40 °C and stir for 5 minutes. Cool the reaction to -78 °C. Slowly add INT1b (16 g, 71.4 mmol) dissolved in 60 ml of 1,4-dioxane solution, and maintain the reaction at -78 °C with stirring for one small-scale test. After the reaction is complete as monitored by TLC, quench the reaction solution by slowly adding ice to a saturated ammonium chloride aqueous solution. Extract three times with 150 ml of ethyl acetate, combine the organic phases, dry with anhydrous sodium sulfate, filter, and evaporate to dryness. The crude product was purified by column chromatography to give compound 1c (13.26 g, brown liquid). LC-MS: ESI [M+H] + =323.3.

[0123] Step 3: Compound 1c (13.26 g, 41.1 mmol) was added to 100 mL of anhydrous ethanol, followed by Pd / C (1.33 g, 10%). The mixture was stirred overnight at room temperature. After the reaction was complete as monitored by LC-MS, the reaction solution was filtered, and the residue was washed with a large amount of ethanol. The filtrates were combined and concentrated by rotary evaporation. A 1,4-dioxane solution (50 mL) of 4 mol / L hydrochloric acid was added, and the mixture was stirred at room temperature for 30 minutes. Diethyl ether was added, precipitating a large amount of solid. The solid was filtered and dried to obtain compound 1d (7.32 g, white solid). LC-MS: ESI [M+H] + =249.3; 1 H NMR (400MHz, DMSO-d6) δ10.44(s,1H),8.62(d,J=1.9Hz,1H),7.75(d,J=1.9Hz,1H),4.34(q,J=7.1Hz,2H),3.87(s,0H),3.24(dd,J= 16.8, 6.3Hz, 1H), 2.97 (dd, J = 16.8, 10.1Hz, 1H), 1.81–1.65 (m, 1H), 1.52–1.36 (m, 1H), 1.32 (t, J = 7.1Hz, 3H), 0.93 (t, J = 7.4Hz, 3H).

[0124] Step 4: Add compound 1d (7.32 g, 29.5 mmol) to a 250 ml reaction flask, then add 120 ml of 1,4-dioxane, followed by DDQ (7.38 g, 32.5 mmol). Reflux overnight. After the reaction is complete as monitored by LC-MS, concentrate the reaction solution by rotary evaporation, add saturated sodium bicarbonate aqueous solution, stir for 1 hour, filter, wash the residue with water, then wash with a small amount of diethyl ether, and dry to obtain compound 1e (2.31 g, yellow solid). LC-MS: ESI [M+H] + =247.3.

[0125] Step 5: Add compound 1e (2.0 g, 8.1 mmol) to a 150 mL reaction flask, then add 60 mL of tetrahydrofuran. Cool to 0 °C, then add a 2.5 mol / L lithium aluminum hydride tetrahydrofuran solution (6.48 mL, 16.2 mmol). React at 0 °C for 2 hours. After the reaction is complete as monitored by TLC, quench the reaction with 5 mL of water, add a large amount of anhydrous sodium sulfate to dry, filter, wash the residue with a large amount of dichloromethane, combine the filtrates, concentrate by rotary evaporation, and dry to obtain compound 1f (1.2 g, white solid). LC-MS: ESI [M+H] + =205.3; 1 H NMR (400MHz, DMSO-d6) δ11.87(s,1H),8.03(d,J=2.0Hz,1H),7.36(d,J=1.0Hz,1H),7 .34(dd,J=2.0,0.9Hz,1H),4.51(s,2H),2.52(d,J=1.8Hz,1H),1.15(t,J=7.4Hz,3H).

[0126] Step 6: Add compound 1f (0.82 g, 4.0 mmol) to a 50 mL reaction flask, along with 20 mL of dichloromethane and 1 mL of N,N-dimethylformamide. Cool to 0 °C, and add sulfoxide (0.87 mL, 12 mmol) dropwise. React at 0 °C for 1 hour. After the reaction is complete as monitored by TLC, concentrate the reaction solution by rotary evaporation. The crude product is purified by column chromatography to give compound 1 g (0.66 g, gray solid). LC-MS: ESI [M+H] + =223.7. 1 H NMR (400MHz, DMSO) δ12.16 (s, 1H), 8.55 (s, 1H), 7.97–7.73 (m, 2H), 4.95 (s, 2H), 2.56 (d, J = 7.4Hz, 2H), 1.19 (t, J = 7.4Hz, 3H).

[0127] Step 7: Compound int-1 (0.05 g, 0.23 mmol), 1 g (0.06 g, 0.28 mmol), N,N-diisopropylethylamine (0.15 g, 1.15 mmol), and potassium iodide (0.19 g, 1.15 mmol) were added to 10 mL of anhydrous acetonitrile. The mixture was stirred at 80 °C for 2 h. After the reaction was complete as monitored by TLC, the reaction solution was concentrated under reduced pressure. The crude product was purified by column chromatography to obtain compound 1 (0.02 g, white solid); LC-MS: ESI [M+H] + =404.5; 1 H NMR (400MHz, DMSO): 11.85(s,1H),8.71(d,J=5.0Hz,1H),8.69(s,1H),8.41(d,J=1.3Hz,1H),8.06–7.91(m,2H),7.75(s,1H),7.6 4(s,1H),6.42(s,1H),3.72(s,2H),3.16(s,2H),2.81(d,J=4.8Hz,3H),2.70(s,2H),2.54(d,J=7.4Hz,4H),1.18(t,J=7.4Hz,3H).

[0128] Example 2: 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthid-3-yl)methyl)-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0129]

[0130] Step 1: Weigh int-1c (1.4 g, 4.5 mmol), dissolve in 20 mL MeOH, add 5 mL water, add lithium hydroxide (570 mg, 13.5 mmol), react at room temperature for 12 h, monitor the reaction by TLC, after the reaction is complete, add 2 M HCl to adjust the pH to 6, add (3 x 25 mL) EA for extraction, combine the organic phases, dry with anhydrous sodium sulfate, and evaporate under vacuum to obtain product 2a (460 mg, pale yellow solid).

[0131] Step 2: Weigh 2a (230 mg, 0.7 mmol), EDCI (191 mg, 1 mmol), HOBT (100 mg, 1 mmol), 2 mL LDM, N-methylmorpholine (250 mg, 2.8 mmol), and d3-methylamine hydrochloride (49 mg, 0.7 mmol). React at room temperature for 12 h, monitor with TLC. After the raw materials are completely consumed, dilute with water, extract with EA, wash the organic phase with water 5 times, dry with anhydrous sodium sulfate, and evaporate to dryness to obtain crude product 2b (150 mg, pale yellow oily liquid).

[0132] Step 3: The crude product of 2b (220 mg, 0.7 mmol) was dissolved in 4 mL of MeOH, followed by the addition of 1.1 mL of 4 M HCl in dioxane solution. The reaction was carried out at room temperature for 12 h, and the reaction was monitored by TLC. After the reaction was complete, potassium carbonate was added, and the mixture was stirred for 30 minutes. The potassium carbonate was then removed by filtration to obtain the crude product of compound 2c (270 mg, yellow-green solid).

[0133] Step 4: Weigh 1f (40.6 mg, 0.2 mmol), add 1 mL of DCM and DMF (15 mg, 0.02 mmol), add thionyl chloride (70 mg, 0.6 mmol) at 0 °C, react for 30 minutes, then react at room temperature for 30 minutes. Monitor by TLC. After the starting material is completely consumed, evaporate to dryness under vacuum. Then add 2c (44.2 mg, 0.2 mmol), DIPEA (180 mg, 1.4 mmol), and KI (10 mg, 0.06 mmol), followed by 2 mL of acetonitrile. Heat to 80 °C and react for 2 hours. Monitor the reaction by TLC. After the reaction is complete, add 2 mL of saturated sodium bicarbonate solution, extract with EA (2 x 3 mL), combine the organic phases, and prepare TLC purified to obtain 2 (12 mg, pale yellow solid). LC-MS: ESI [M+H] + =407.5. 1 H NMR (400MHz, DMSO): 11.85(s,1H),8.71(d,J=5.0Hz,1H),8.69(s,1H),8.41(d,J=1.3Hz,1H),8.06–7.91(m,2H),7.75( s,1H),7.64(s,1H),6.42(s,1H),3.72(s,2H),3.16(s,2H),2.70(s,2H),2.54(d,J=7.4Hz,4H),1.18(t,J=7.4Hz,3H).

[0134] Example 3: 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl-d)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0135]

[0136] Step 1: Compound 1f (0.4 g, 2.0 mmol) and dichloromethane (20 mL) were added to a 50 mL reaction flask. After cooling to 0 °C, Dysmartin oxidant (1.7 g, 4.0 mmol) was added. The reaction was carried out at 0 °C for 1.5 h. After the reaction was complete as monitored by TLC, the mixture was filtered through diatomaceous earth. The residue was washed with dichloromethane, and the filtrate was concentrated by rotary evaporation and purified by column chromatography to obtain compound 3a (0.38 g, pale yellow solid). LC-MS: ESI [M+H] + =203.2; 1 H NMR (400MHz, DMSO-d6) δ12.17(s,1H),10.16(s,1H),8.92(d,J=1.8Hz,1H),7.85–7.83(m,2H),2.59(qd,J=7.4,1.3Hz,2H),1.21(t,J=7.4Hz,3H).

[0137] Step 2: Weigh compound 3a (0.2 g, 1 mmol) into a reaction flask, add 15 mL of diethyl ether and 4 mL of methanol, cool to 0 °C, add sodium borodeuteride (46 mg, 1.1 mmol), and react at room temperature for 1 hour. After the reaction is complete as detected by TLC, quench with a small amount of water, concentrate by rotary evaporation, add a small amount of ice water to form a slurry, filter, and dry. Compound 3b (110 mg, white solid) is obtained; LC-MS: ESI [M+H] + =206.2.

[0138] Step 3: Compound 3b (110 mg, 0.5 mmol) was added to a 25 ml reaction flask, along with 10 ml of dichloromethane and 0.1 ml of N,N-dimethylformamide. The mixture was cooled to 0 °C, and thionyl chloride (88 mg, 0.75 mmol) was added dropwise. The reaction was carried out at 0 °C for 1 hour. After the reaction was completed as monitored by TLC, the reaction solution was concentrated by rotary evaporation. The crude product was purified by column chromatography to obtain compound 3c (103 mg, white solid); LC-MS: ESI [M+H] + =224.7.

[0139] Step 4: Compound 3c (67 mg, 0.3 mmol), int-1 (96 mg, 0.33 mmol), N,N-diisopropylethylamine (194 mg, 1.5 mmol), and potassium iodide (5 mg, 0.03 mmol) were added to 10 mL of anhydrous acetonitrile. The mixture was stirred at 80 °C for 2 hours. After the reaction was complete as monitored by TLC, the reaction solution was concentrated under reduced pressure. The crude product was purified by column chromatography to obtain compound 3 (60 mg, pale yellow solid); LC-MS: ESI [M+H] + =405.5; 1H NMR (400MHz, DMSO) δ11.85(s,1H),8.70(s,2H),8.42(d,J=1.7Hz,1H),7.99(d,J=2.0Hz,2H),7.76(s,1H),7.65(s,1H) ,6.43(s,1H),3.69(s,1H),3.17(s,2H),2.82(d,J=4.8Hz,3H),2.71(s,2H),2.60–2.53(m,4H),1.19(t,J=7.4Hz,3H).

[0140] Example 4: 1'-((2-ethyl-5-fluoro-3-oxo-3,4-dihydroquinoxalin-6-yl)methyl)-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0141]

[0142] Step 1: Weigh compound 4a (50 g, 314 mmol) and N-bromosuccinimide (67 g, 377 mmol) into a reaction flask, add 300 mL of sulfuric acid, and heat to 80 °C and react overnight. After the reaction is complete as detected by TLC, cool to room temperature, slowly add ice water to dilute, extract three times with ethyl acetate, combine the organic phases, wash with water, wash with saturated sodium bicarbonate solution, dry with anhydrous sodium sulfate, filter, and evaporate to dryness. The crude product obtained is purified by column chromatography to give compound 4b (65 g, pale yellow solid); LC-MS: ESI [M+H] + =238.9.

[0143] Step 2: Compound 4b (43 g, 181 mmol), compound 4c (21 g, 181 mmol), N,N-diisopropylethylamine (70 g, 543 mmol), and N,N-dimethylformamide (100 ml) were added to a 500 ml reaction flask and stirred overnight at room temperature. After the reaction was complete as monitored by TLC, the reaction solution was concentrated under reduced pressure, extracted three times with ethyl acetate and water, and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography to give compound 4d (37 g, orange-red solid); LC-MS: ESI [M+H] + =336.1.

[0144] Step 3: Compound 4d (26 g, 78 mmol) was added to 200 ml of anhydrous methanol and 5 ml of water, followed by ammonium chloride (35 g, 621 mmol). The mixture was cooled to 0 °C, and zinc powder (43 g, 621 mmol) was slowly added. The mixture was then heated to room temperature and reacted for 1 hour. After the reaction was complete as monitored by TLC, the mixture was filtered. The filtrate was concentrated under reduced pressure, and 30 ml of a 4 mol / L dioxane solution was added. The mixture was reacted at room temperature for 1 hour. After the reaction was complete as monitored by TLC, petroleum ether was added to precipitate a solid. The solid was filtered, dried, and compound 4e (16 g, grayish-white solid) was obtained. LC-MS: ESI [M+H] + =274.1.

[0145] Step 4: Compound 4e (16 g, 57 mmol) and 2,3-dichloro-5,6-dicyanobenzoquinone (16 g, 69 mmol) were added to 500 mL of dichloromethane and reacted overnight at room temperature. After the reaction was complete as monitored by TLC, the reaction solution was concentrated under reduced pressure, quenched with saturated sodium bicarbonate aqueous solution, and extracted three times with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography to give compound 4f (7.9 g, brown solid); LC-MS: ESI [M+H] + =272.1.

[0146] Step 5: Weigh compound 4f (0.5 g, 1.8 mmol), tributyltin methanol (0.65 g, 2.0 mmol), and Xphos-Pd-G2 (73 mg, 0.09 mmol), add 15 mL of dioxane, replace with nitrogen, heat to 80 °C and react overnight. After the reaction is complete as monitored by TLC, extract three times with water and ethyl acetate. Combine the organic phases, dry with anhydrous sodium sulfate, filter, and evaporate to dryness. The crude product is purified by column chromatography to give compound 4 g (0.4 g, pale yellow solid); LC-MS: ESI [M+H] + =223.2.

[0147] Step 6: 4 g (0.4 g, 0.8 mmol) of compound was added to 10 mL of hydrobromic acid aqueous solution, and the mixture was heated to 80 °C and reacted for 3 hours. After the reaction was complete as monitored by TLC, saturated sodium bicarbonate aqueous solution was added to quench the reaction. The mixture was extracted three times with dichloromethane, and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography to give compound 4h (0.24 g, pale yellow solid); LC-MS: ESI [M+H] + =286.1.

[0148] Step 7: Compound 4h (57 mg, 0.2 mmol), 2c (64 mg, 0.22 mmol), N,N-diisopropylethylamine (0.13 g, 1.0 mmol), and potassium iodide (3 mg, 0.02 mmol) were added to 10 mL of anhydrous acetonitrile. The mixture was stirred at 85 °C for 2 h. After the reaction was complete as monitored by TLC, the reaction solution was concentrated under reduced pressure. The crude product was purified by column chromatography to obtain compound 4 (16 mg, pale yellow solid). LC-MS: ESI [M+H] + =425.2; 1 H NMR (400MHz, DMSO) δ12.42(s,1H),8.75–8.63(m,2H),8.03–7.91(m,2H),7.55(d,J=8.3Hz,1H),7.36–7. 25(m,1H),6.41(s,1H),3.76(s,2H),2.89–2.76(m,4H),2.72(t,J=5.5Hz,2H),2.54(s,2H),1.22(s,3H).

[0149] Example 5: N-cyclopropyl-1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl-d)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0150]

[0151] Step 1: Weigh 2a (180 mg, 0.6 mmol), EDCI (144 mg, 0.75 mmol), HOBT (100 mg, 0.75 mmol), 2 mL DMF, N-methylmorpholine (290 mg, 3.2 mmol), and cyclopropylamine (34 mg, 0.6 mmol). React at low temperature for 12 h, monitor with TLC. After the starting materials are completely consumed, dilute with water, extract with EA, wash the organic phase with water 5 times, dry with anhydrous sodium sulfate, and evaporate to dryness to obtain crude product 5a (180 mg, yellow oily liquid).

[0152] Step 2: The crude product of 5a (180 mg, 0.6 mmol) was dissolved in 5 mL of MeOH, followed by the addition of 0.9 mL of 4 M HCl in dioxane solution. The reaction was carried out at room temperature for 12 h, and the reaction was monitored by TLC. After the reaction was complete, potassium carbonate was added, and the mixture was stirred for 30 minutes. The potassium carbonate was then removed by filtration to obtain the crude product of compound 5b (180 mg, yellowish-brown solid).

[0153] Step 3: Weigh 3b (40.6 mg, 0.2 mmol), add 1 mL of DCM and DMF (15 mg, 0.02 mmol), add thionyl chloride (70 mg, 0.6 mmol) at 0 °C and react for 30 minutes. Then, react at room temperature for 30 minutes, monitoring with TLC. After the starting material is completely consumed, evaporate to dryness under vacuum. Then add 5b (44.2 mg, 0.2 mmol), DIPEA (180 mg, 1.4 mmol), and KI (10 mg, 0.06 mmol), followed by 2 mL of acetonitrile. Heat to 80 °C and react for 2 hours. Monitor the reaction with TLC. After the reaction is complete, add 2 mL of saturated sodium bicarbonate solution, extract with EA (2 x 3 mL), combine the organic phases, and prepare TLC purified to obtain 5 (8 mg, pale yellow solid). LC-MS: ESI [M+H] + =431.2. 1 H NMR (400MHz, DMSO) δ11.87(s,1H),8.69-8.65(m,2H),8.43(s,1H),8.04-7.94(m,2H),7.76(s,1H),7.66(s,1H),6.41(s,1H),3.76-3.66(m,1 H),3.17(s,2H),2.95-2.85(m,1H),2.77-2.68(m,3H),2.55(q,J=13.2Hz,2H),2.36-2.33(m,1H),1.19(t,J=13.2Hz,3H),0.75-0.61(m,4H).

[0154] Example 6: 1'-((7-(ethyl-d5)-6-oxo-5,6-dihydro-1,5-naphthid-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0155]

[0156] Step 1: Weigh 16 g (406 mmol) of 60% sodium hydride, add 200 mL of DME, purge with nitrogen three times, cool to 0 °C, and add compound 6a (76 g, 338 mmol) dropwise. Incubate the reaction at room temperature for 2 hours, add 50 g (439 mmol) of deuterated bromoethane, and incubate at 60 °C for 3 hours. After monitoring the reaction progress by HPLC, slowly quench the reaction mixture in ice water. Extract three times with ethyl acetate, combine the organic phases, dry with anhydrous sodium sulfate, filter, and evaporate to dryness. The crude compound 6c (83 g, colorless liquid) is obtained.

[0157] Step 2: Add 60% sodium hydride (13g, 322mmol) to the reaction flask, add 200ml of tetrahydrofuran, and then purge with nitrogen three times. Cool to 0℃, slowly add compound 6c (83g, 322mmol), stir at 0℃ for 10 minutes, heat to room temperature and stir for 10 minutes, then heat to 40℃ and stir for 5 minutes, and cool to -78℃. Slowly add a solution of compound 1b (48g, 215mmol) dissolved in 200ml of tetrahydrofuran, and maintain at -78℃ with stirring for 1 hour. After the reaction is complete as monitored by TLC, quench the reaction solution slowly in ice-cold saturated ammonium chloride aqueous solution, extract three times with ethyl acetate, combine the organic phases, dry with anhydrous sodium sulfate, filter, and evaporate to dryness. The crude product is purified by column chromatography to obtain compound 6d (41g, yellow-green liquid). LC-MS: ESI [M+H] + =328.4.

[0158] Step 3: Compound 6d (1.0 g, 3.0 mmol) was added to 10 mL of anhydrous ethanol, followed by the addition of Pd / C (0.1 g, 10%). Hydrogen was then introduced three times, and the mixture was stirred overnight at room temperature. After the reaction was complete as monitored by LC-MS, the reaction solution was filtered, and the residue was washed with copious amounts of ethanol. The filtrates were combined and concentrated by rotary evaporation. A 1,4-dioxane solution (12 mL) in 4 mol / L hydrochloric acid was added, and the mixture was stirred at room temperature for 30 minutes. Diethyl ether was added, precipitating a large amount of solid. The solid was filtered and dried to obtain compound 6e (0.51 g, white solid). LC-MS: ESI [M+H] + =254.3.

[0159] Step 4: Add compound 6e (0.51 g, 2.0 mmol) to a 50 mL reaction flask, then add 15 mL of 1,4-dioxane, followed by DDQ (0.50 g, 2.2 mmol). Reflux overnight. After the reaction is complete as monitored by LC-MS, concentrate the reaction solution by rotary evaporation, add saturated sodium bicarbonate aqueous solution, stir for 1 hour, filter, wash the residue with water, then wash with a small amount of diethyl ether, and dry to obtain compound 6f (0.3 g, yellow solid). LC-MS: ESI [M+H] + =252.3.

[0160] Step 5: Add compound 6f (0.3 g, 1.2 mmol) to a reaction flask, add 15 mL of tetrahydrofuran, cool to 0 °C, then add 1.44 mL of a 2.5 mol / L lithium aluminum hydride tetrahydrofuran solution (3.6 mmol), and react at 0 °C for 2 hours. After the reaction is complete as monitored by TLC, quench the reaction with 1 mL of water, add a large amount of anhydrous sodium sulfate to dry, filter, wash the residue with a large amount of dichloromethane, combine the filtrates, concentrate by rotary evaporation, and dry to obtain compound 6 g (0.2 g, yellow solid). LC-MS: ESI [M+H]+ =210.3.

[0161] Step 6: Add 6 g (0.2 g, 0.96 mmol) of compound to a 25 mL reaction flask, along with 10 mL of dichloromethane and 0.1 mL of N,N-dimethylformamide. Cool to 0 °C, and add 0.35 g (2.9 mmol) of thionyl chloride dropwise. React at 0 °C for 1 hour. After the reaction is complete as monitored by TLC, concentrate the reaction solution by rotary evaporation. The crude product is purified by column chromatography to obtain compound 6h (0.2 g, gray solid). LC-MS: ESI [M+H] + =228.7.

[0162] Step 7: Compound 6h (0.2 g, 0.88 mmol), int-1 (0.25 g, 0.88 mmol), N,N-diisopropylethylamine (0.45 g, 3.52 mmol), and potassium iodide (15 mg, 0.09 mmol) were added to 10 mL of anhydrous acetonitrile. The mixture was stirred at 85 °C for 2 hours. After the reaction was complete as monitored by TLC, the reaction solution was concentrated under reduced pressure. The crude product was purified by column chromatography to obtain compound 6 (160 mg, white solid). LC-MS: ESI [M+H] + =409.5; 1 H NMR (400MHz, CDCl3) δ10.97(s,1H),8.57(d,J=2.0Hz,1H),8.54(d,J=1.7Hz,1H),8.15(d,J=8.2Hz,1H),7.96(d,J=4.9Hz,1H),7.86(s,1H),7.80(dd, J=8.2,2.3Hz,1H),7.66(s,1H),6.23(s,1H),3.78(s,2H),3.27(d,J=3.1H z, 2H), 3.04 (d, J = 5.1Hz, 3H), 2.79 (t, J = 5.6Hz, 2H), 2.60 (d, J = 1.4Hz, 2H).

[0163] Example 7: 1'-((7-(ethyl-d5)-6-oxo-5,6-dihydro-1,5-naphthid-3-yl)methyl)-N,2-dimethyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0164]

[0165] Step 1: Compound 7a (0.5 g, 2.2 mmol), int-1b (2.4 g, 5.5 mmol), Pd(dppf)Cl2 (0.16 g, 0.22 mmol), and potassium carbonate (0.7 g, 5.5 mmol) were added to a mixed solvent of 7 ml dioxane, 3 ml anhydrous ethanol, and 4 ml water. The mixture was then purged with nitrogen three times and reacted at 90 °C for 2 h under nitrogen protection. After TLC detection of complete reaction, the reaction mixture was cooled to room temperature, and 30 ml dichloromethane and 20 ml water were added. The mixture was separated into layers in a separatory funnel. The aqueous phase was extracted twice with dichloromethane. The combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography to obtain compound 7b (0.7 g, white solid).

[0166] Step 2: Compound 7b (0.7 g, 26 mmol) was added to 10 ml of anhydrous methanol, followed by 20 ml of methylamine aqueous solution. The mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain compound 7c (0.5 g, white solid).

[0167] Step 3: Compound 7c (0.5 g, 1.7 mmol) was added to 10 ml of anhydrous methanol, followed by 10 ml of 4 mol / L dioxane hydrochloride solution. The mixture was stirred at room temperature for 0.5-1 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain compound 7d (0.3 g, white solid).

[0168] Step 4: Compounds 7d (0.05 g, 0.25 mmol), 6h (0.071 g, 0.231 mmol), N,N-diisopropylethylamine (0.17 g, 1.25 mmol), and potassium iodide (0.22 g, 1.25 mmol) were added to 10 mL of anhydrous acetonitrile. The mixture was stirred at 80 °C for 2 h. After the reaction was complete as monitored by TLC, the reaction solution was concentrated under reduced pressure. The crude product was purified by column chromatography to obtain compound 7 (0.024 g, white solid). LC-MS: ESI [M+H] + =423.2. 1H NMR (400MHz, Chloroform-d) δ11.90(s,1H),8.50(d,J=2.0Hz,1H),7.99(d,J=5.0Hz,1H),7.91(d,J=7.8Hz,1H),7.81(s,1H),7.67(d,J=1.9Hz,1H),7.4 6(d,J=7.8Hz,1H),5.64-5.50(m,1H),3.73(s,2H),3.15(d,J=3.1Hz,2H),2 .96(d,J=5.1Hz,3H),2.69(t,J=5.5Hz,2H),2.47(s,3H),2.36-2.32(m,2H).

[0169] Example 8: 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-2-methyl-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0170]

[0171] The preparation of 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-2-methyl-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was performed according to Example 2. LC-MS: ESI [M+H] + =421.2. 1 H NMR (400MHz, DMSO-d6) δ11.86(s,1H),8.56(s,1H),8.43(d,J=1.9Hz,1H),7.80(d,J=7.8Hz,1H),7.76(s,1H),7.70-7.60(m,2H) ,5.70(s,1H),3.73(s,2H),3.12(d,J=3.1Hz,2H),2.69(t,J=5.5Hz,2H),2.57-2.53(m,5H),2.36(s,2H),1.18(t,J=7.4Hz,3H).

[0172] Example 9: 1'-((7-(ethyl-d5)-6-oxo-5,6-dihydro-1,5-naphthid-3-yl)methyl)-2-fluoro-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0173]

[0174] Step 1: Compound int-1a (3.7 g, 17.1 mmol) and silver fluoride (10 g, 68.4 mmol) were added to 40 mL of anhydrous acetonitrile. The mixture was then purged with nitrogen three times and stirred at room temperature for 48 h under nitrogen protection. After the reaction was confirmed to be complete by TLC, the reaction mixture was concentrated to dryness under reduced pressure. The crude product was purified by column chromatography to obtain compound 9a (2.2 g, white solid).

[0175] Step 2: Compound 9a (1 g, 4.3 mmol), int-1b (1.59 g, 5.1 mmol), Pd(dppf)Cl2 (0.31 g, 0.43 mmol), and potassium carbonate (1.5 g, 10.8 mmol) were added to a mixed solvent of 7 ml dioxane, 3 ml anhydrous ethanol, and 4 ml water. The mixture was then purged with nitrogen three times and reacted at 90 °C for 2 h under nitrogen protection. After TLC detection of complete reaction, the reaction mixture was cooled to room temperature, and 30 ml dichloromethane and 20 ml water were added. The mixture was separated into layers in a separatory funnel. The aqueous phase was extracted twice with dichloromethane. The combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography to give compound 9b (1.2 g, white solid).

[0176] Step 3: Add 9b (1g, 2.9mmol), aqueous methylamine solution (5g, 161.3mmol), and anhydrous methanol (20ml) to a 100ml reaction flask and stir overnight at room temperature. After the reaction is complete as monitored by TLC, concentrate the reaction solution under reduced pressure to dryness to obtain compound 9c (0.8g, white solid).

[0177] Step 4: Compound 9c (0.5 g, 1.4 mmol) was added to 10 ml of anhydrous methanol, followed by 10 ml of 4 mol / L dioxane hydrochloride solution. The mixture was stirred at room temperature for 0.5-1 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain compound 9d (0.3 g, white solid).

[0178] Step 5: Compounds 9d (0.05 g, 0.21 mmol), 6h (0.056 g, 0.26 mmol), N,N-diisopropylethylamine (0.14 g, 1.05 mmol), and potassium iodide (0.17 g, 1.05 mmol) were added to 10 mL of anhydrous acetonitrile. The mixture was stirred at 80 °C for 2 h. After the reaction was complete as monitored by TLC, the reaction solution was concentrated under reduced pressure. The crude product was purified by column chromatography to obtain compound 9 (0.02 g, white solid). LC-MS: ESI [M+H] + =427.2. 1H NMR (400MHz, Chloroform-d) δ11.47(s,1H),8.48(d,J=1.8Hz,1H),7.99(dd,J=7.7,1.7Hz,1H),7.84-7.72(m,2H),7.64-7. 52(m,2H),6.17-6.08(m,1H),3.71(s,2H),3.20(d,J=3.3Hz,2H),2.95(d,J=5.0Hz,3H),2.70(t,J=5.6Hz,2H),2.51(s,2H).

[0179] Example 10: 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-2-fluoro-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0180]

[0181] The preparation of 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-2-fluoro-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was performed according to Example 2. LC-MS: ESI [M+H] + =425.2. 1 H NMR(400MHz,Chloroform-d)δ11.15(s,1H),8.47(d,J=1.8Hz,1H),7.99(dd,J=7.7,1.7Hz,1H),7.83-7.73(m,2H),7.63(s,1H) ,7.57(s,1H),6.13(t,J=2.6Hz,1H),3.72(s,2H),3.21(d,J=3.3Hz,2H),2.75-2.62(m,4H),2.52(s,2H),1.24(t,J=7.4Hz,3H).

[0182] Example 11: 1'-((7-(ethyl-d5)-6-oxo-56-dihydro-1,5-naphthid-3-yl)methyl)-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0183]

[0184] The preparation of 1'-((7-(ethyl-d5)-6-oxo-56-dihydro-1,5-naphthid-3-yl)methyl)-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was performed according to Example 1. LC-MS: ESI [M+H] + =412.3. 1 HNMR(400MHz,Chloroform-d)δ11.25(s,1H),8.49(dd,J=8.7,2.0Hz,2H),8.08(d,J=8.2Hz,1H),7.87(s,1H),7.79(s,1H),7.73(dd,J =8.2, 2.3Hz, 1H), 7.62 (d, J = 1.7Hz, 1H), 6.15 (t, J = 3.7Hz, 1H), 3.72 (s, 2H), 3.20 (d, J = 3.2Hz, 2H), 2.73 (t, J = 5.6Hz, 2H), 2.54 (s, 2H).

[0185] Example 12: N-cyclopropyl-1'-((7-(ethyl-d5)-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-1'-,2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0186]

[0187] The preparation of N-cyclopropyl-1'-((7-(ethyl-d5)-6-oxo-5,6-dihydro-1,5-naphthylpyridin-3-yl)methyl)-1'-,2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was performed according to Example 1. LC-MS: ESI [M+H] + =435.3. 1 H NMR(400MHz,Chloroform-d)δ11.53(s,1H),8.55-8.36(m,2H),8.08(dd,J=8.3,0 .8Hz,1H),7.92(d,J=3.8Hz,1H),7.80(s,1H),7.72(dd,J=8.2,2.3Hz,1H),7.63(d ,J=1.9Hz,1H),6.23-6.06(m,1H),3.72(s,2H),3.20(d,J=3.1Hz,2H),2.91-2.84 (m,1H),2.72(t,J=5.7Hz,2H),2.53(s,2H),0.86-0.72(m,2H),0.63-0.53(m,2H).

[0188] Example 13: 1'-((2-(ethyl-d5)-5-fluoro-3-oxo-3,4-dihydroquinoxalin-6-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0189]

[0190] Step 1: Weigh 13a (10.0 g, 39.5 mmol) into a reaction flask, add 100 mL of DMF to dissolve it, and add t-BuOK (6.6 g, 59.2 mmol) in portions to the system at -10 °C under nitrogen protection. After the addition is complete, continue stirring at -10 °C for 1 h. Then, slowly add deuterated bromoethane (5.0 g, 43.4 mmol) dropwise. After the addition is complete, return to room temperature and stir for 3 h. Add 300 mL of water to the system, extract with EA (500 mL × 3), combine the organic phases and wash with saturated brine. Separate the layers, dry the organic phase with anhydrous Na2SO4, filter, and concentrate under vacuum to obtain a yellow oily liquid 13b (11.0 g, 97%). LC-MS: ESI [M+H] + =287.2.

[0191] Step 2: Weigh 13b (11.0 g, 38.4 mmol), add 120 mL of a 3 / 1 mixture of EA and H2O, add 28.8 mL of 4 M ethyl acetate hydrochloride solution (115.2 mmol), stir overnight, concentrate under vacuum to remove the solvent, and slurry with EA to obtain a white solid 13c hydrochloride (2.5 g, 41%). LC-MS: ESI [M+H] + =123.1.

[0192] Step 3: Weigh 2.5 g (15.8 mmol) of 13c hydrochloride into a reaction flask, dissolve it in 30 mL of DMF, add 8.1 g (63.0 mmol) of DIEA, and add 4b (4.1 g (17.3 mmol) while stirring at room temperature. Stir at room temperature for 5 h under nitrogen protection. After the reaction is complete, add 90 mL of pure water and extract with EA (150 mL × 2). Combine the organic phases and wash with saturated brine. Separate the layers and concentrate the organic phase under vacuum. Filter the column with PE and DCM (0-20%) to obtain a yellow oily liquid 13d (4.8 g, 90%). LC-MS: ESI [M+H] + =340.0.

[0193] Step 4: Dissolve 13d (4.5 g, 13.2 mmol) in 80 mL of glacial acetic acid. Slowly add reduced Fe (4.4 g, 79.4 mmol) powder to the system while stirring at room temperature. After the addition is complete, stir at 70 °C for 1 h. Filter while hot, wash the filter cake with a mixed solvent of DCM and MeOH, concentrate the filtrate under vacuum, and purify by column chromatography with PE and EA (0-25%) to obtain a white solid 13e (2.1 g, 57%). LC-MS: ESI [M+H] + =278.0.

[0194] Step 5: Weigh 13e (2.1 g, 7.6 mmol) into a reaction flask, add 30 mL of DCM and stir. Then slowly add DDQ (2.1 g, 9.1 mmol). After the addition is complete, stir overnight at room temperature. After the reaction is complete, add saturated NaHCO3 aqueous solution to the system, separate the layers, and extract the aqueous phase with a 5:1 mixture of DCM and MeOH (30 mL × 3). Combine the organic phases and dry them with anhydrous Na2SO4. Filter and concentrate to dryness. Purify by column chromatography with DCM and MeOH (0-3%) to obtain a white solid 13f (2.0 g, 96%). LC-MS: ESI [M+H] + =276.0.

[0195] Step 6: Weigh 13f (1.2 g, 5.3 mmol), (tributyltin)methanol (2.8 g, 8.7 mmol), and XPos-Pd-G2 (416.4 mg, 0.4 mmol) into a reaction flask. Add 30 mL of dioxane to purge nitrogen 3-5 times. Then, stir at 90 °C for 4 h under nitrogen protection. After the reaction is complete, concentrate the solvent under vacuum and purify by column chromatography with DCM and MeOH (0-5%) to obtain 13 g (946.0 mg, 76%) of white solid. LC-MS: ESI [M+H] + =228.1; 1 H NMR (400MHz, DMSO) δ 12.46 (s, 1H), 7.59 (d, J = 8.3Hz, 1H), 7.37 (t, J = 7.7Hz, 1H), 5.46 (t, J = 5.8Hz, 1H), 4.68 (d, J = 5.6Hz, 2H).

[0196] Step 7: Weigh 13g (650mg, 2.9mmol) into a reaction flask, add 20mL of DCM to dissolve, and slowly add Dess-Martin oxidant (1.5g, 3.4mmol) at 0℃. After the addition is complete, maintain 0℃ and continue stirring for 0.5h. After the reaction is complete, add saturated NaHCO3 aqueous solution to the system, stir for 10min, and then separate the layers. Concentrate the organic solvent to dryness under vacuum, and purify by column chromatography with DCM and MeOH (0-5%) to obtain a white solid of 13h (610mg, 95%). LC-MS: ESI [M+H] + =226.1; 1 H NMR (400MHz, CDCl3) δ10.37(s,1H),9.85(s,1H),7.72–7.63(m,2H).

[0197] Step 8: Weigh int-1a (99.2 mg, 0.34 mmol) into a reaction flask, add 5 mL of DCM, then add TEA to release it, then add glacial acetic acid dropwise to adjust the pH to approximately 5, and finally add 13h (70.0 mg, 0.31 mmol) and stir at room temperature for 2 h. Add sodium triacetoxyborohydride (263.5 mg, 1.24 mmol) to the system and continue stirring for 2 h. Add saturated NH4Cl aqueous solution and stir. Separate the liquid and aqueous phases, extract with a 5:1 mixture of DCM and MeOH (30 mL × 3), combine the organic phases and dry with anhydrous Na2SO4, filter and concentrate, prepare by HPLC and freeze-dry to obtain white solid 13 (16 mg, 12%). LC-MS: ESI [M+H] + =427.2; 1 H NMR (400MHz, DMSO) δ12.37(s,1H),8.76–8.66(m,2H),8.02–7.94(m,2H),7.56(d,J=8.3Hz,1H),7.32(t,J=7 .6Hz,1H),6.42(s,1H),3.77(s,2H),3.18(s,2H),2.82(d,J=4.7Hz,3H),2.73(t,J=5.4Hz,2H),2.55(s,2H).

[0198] Example 14: 1'-((2-(ethyl-d5)-5-fluoro-3-oxo-3,4-dihydroquinoxalin-6-yl)methyl)-2-fluoro-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0199]

[0200] Weigh 9d (150.5 mg, 0.49 mmol) into a reaction flask, add 5 mL of DCM, then add TEA to release it, then add glacial acetic acid dropwise to adjust the pH to approximately 5, and finally add 13h (100.0 mg, 0.44 mmol) and stir at room temperature for 2 h. Add sodium triacetoxyborohydride (376.4 mg, 1.78 mmol) to the system, continue stirring for 2 h, then add saturated NH4Cl aqueous solution and stir. Separate the solution, extract the aqueous phase with a 5:1 mixture of DCM and MeOH (30 mL × 3), combine the organic phases and dry with anhydrous Na2SO4, filter and concentrate the filtrate, and prepare the residue by HPLC and freeze-dry to give a white solid 14 (60 mg, 30%). LC-MS: ESI [M+H] + =445.2; 1 H NMR (400MHz, DMSO) δ12.47(s,1H),8.72–8.58(m,1H),8.08(dd,J=9.8,7.8Hz,1H),7.92(dd,J=7.7,1.4Hz,1H),7.56(d,J=8.3Hz,1H),7 .32(t,J=7.7Hz,1H),6.25(s,1H),3.76(s,2H),3.18(d,J=2.5Hz,2H),2.80(d,J=4.8Hz,3H),2.70(t,J=5.5Hz,2H),2.51–2.46(m,2H).

[0201] Example 15: 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl-d2)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0202]

[0203] Step 1: Compound 1e (2.0 g, 8.1 mmol) was added to a 150 mL reaction flask, followed by 60 mL of tetrahydrofuran. The mixture was cooled to 0 °C, and then a 2.5 mol / L solution of lithium aluminum deuteride in tetrahydrofuran (6.48 mL, 16.2 mmol) was added. The reaction was carried out at 0 °C for 2 hours. After the reaction was complete as monitored by TLC, 5 mL of water was added to quench the reaction. The mixture was dried with a large amount of anhydrous sodium sulfate, filtered, and the residue was washed with a large amount of dichloromethane. The filtrates were combined and concentrated by rotary evaporation. After drying, compound 15a (0.8 g, white solid) was obtained. LC-MS: ESI [M+H] + =207.3.

[0204] Step 2: Compound 15a (0.82 g, 4.0 mmol) was added to a 50 mL reaction flask, along with 20 mL of dichloromethane and 1 mL of N,N-dimethylformamide. The mixture was cooled to 0 °C, and sulfoxide (0.87 mL, 12 mmol) was added dropwise. The reaction was carried out at 0 °C for 1 hour. After the reaction was completed as monitored by TLC, the reaction solution was concentrated by rotary evaporation. The crude product was purified by column chromatography to obtain compound 15b (0.52 g, gray solid). LC-MS: ESI [M+H] + =225.7.

[0205] Step 3: Compound 15b (56 mg, 0.25 mmol), INT1 (71 mg, 0.231 mmol), N,N-diisopropylethylamine (0.17 g, 1.25 mmol), and potassium iodide (0.22 g, 1.25 mmol) were added to 10 mL of anhydrous acetonitrile. The mixture was stirred at 80 °C for 2 h. After the reaction was complete as monitored by TLC, the reaction solution was concentrated under reduced pressure. The crude product was purified by column chromatography to obtain compound 15 (25 mg, white solid); LC-MS: ESI [M+H] + =406.4.

[0206] Example 16: 1'-((7-(ethyl-2,2,2-d3)-6-oxo-5,6-dihydro-1,5-naphthid-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0207]

[0208] Step 1: Weigh 16a (500.0 g, 1.99 mol), add 2.5 L of methanol, and cool to 0-5 °C under nitrogen protection with stirring. Weigh sodium methoxide (118.0 g, 2.19 mol), add 1.0 L of methanol, stir to dissolve, and then add dropwise to the reaction system under controlled temperature of 0-5 °C. After the addition is complete, allow it to return to the temperature naturally and stir for 1 h. After the reaction is complete as detected by TLC, add 2.0 L of water to the reaction system and concentrate under reduced pressure at 40-50 °C until no liquid is dispensed. Extract the aqueous layer twice with 4.0 L and 1.0 L of ethyl acetate, respectively. Combine the organic phases and concentrate under reduced pressure at 40-50 °C to constant weight to obtain compound 16b (480 g, white solid).

[0209] Step 2: Weigh 16b (475g, 1.93mol), add 2.85L of DMF and 2.85L of DMF-DMA. After the addition is complete, heat to 100℃ and stir for 2h. After the reaction is complete as detected by TLC, concentrate the reaction solution at 70-80℃ until no liquid is discharged. Then add 10.0L of water, stir at 20-30℃ for 2h, filter, and dry the filter cake under reduced pressure at 60-70℃ to constant weight to obtain compound 16c (612g, red solid).

[0210] Step 3: Weigh 16c (500g, 1.91mol), add 2.56L of tetrahydrofuran, weigh sodium periodate (805.0g, 3.72mol) and dissolve it in 2.56L of water. Add the sodium periodate aqueous solution dropwise to the reaction system while maintaining the temperature at 25-30℃. After the addition is complete, stir at 25-30℃ for 2-4 hours. After the reaction is complete as detected by TLC, add 2.56L of water and 4.0L of ethyl acetate. Stir to separate the layers. Wash the aqueous layer twice with 2.0L of ethyl acetate. Combine the organic layers and wash them successively with 4.0L of saturated sodium thiosulfate solution and 4.0L of saturated saline solution. Separate the layers and concentrate the organic layer under reduced pressure at 40-50℃ to constant weight to obtain compound 16d (500g, oily substance).

[0211] Step 4: Weigh 16d (500.0 g, 1.69 mol), ethyl 3,3-diethoxypropionate (1457.0 g, 7.61 mol), and add 7.5 L of anhydrous ethanol. Add stannous chloride (1815.0 g, 9.57 mol) in portions. After the addition is complete, heat to reflux and react for 1-2 h. After the reaction is complete as detected by TLC, concentrate the reaction solution under reduced pressure at 40-50 °C to constant weight. Add 15.0 L of ethyl acetate, adjust the pH to 7-8 with saturated sodium bicarbonate solution, filter, concentrate the organic layer under reduced pressure at 40-50 °C to constant weight, and purify by column chromatography to obtain compound 16e (230.0 g, white solid).

[0212] Step 5: Weigh 16e (80.0 g, 257.1 mmol), add 960 ml of tetrahydrofuran, cool in an ice-water bath to 0 °C, add diisopropylaluminum hydride (1 M, 771.3 ml) dropwise, and after the addition is complete, warm to 20-25 °C to react. After the reaction is complete as detected by LC-MS, add 800 ml of saturated ammonium chloride solution dropwise to the reaction system, filter, evaporate the filtrate to dryness, add 80 ml of dichloromethane to slurry, filter, and concentrate the filter cake under reduced pressure at 40-50 °C to constant weight to obtain compound 16f (43.0 g, yellow solid).

[0213] Step 6: Weigh 16f (45.0 g, 167.2 mmol), DIPEA (64.8 g, 501.6 mmol), sodium iodide (5.0 g, 33.4 mmol), add 450 ml of tetrahydrofuran, and add MOMCl (26.9 g, 334.4 mmol) dropwise while stirring. After the addition is complete, heat to 45-50 °C and stir for 3 h. After the reaction is complete as detected by TLC, cool to 20-25 °C, add 500 ml of water, separate the layers, extract the aqueous layer with 300 ml of ethyl acetate, combine the organic layers and concentrate to dryness under reduced pressure at 40-50 °C. Column chromatography gives 16 g of compound (34.8 g, yellow solid).

[0214] Step 7: Weigh 16g (39.8g, 127.1mmol), add triethylamine (15.5g, 152.5mmol), Pd(dppf)Cl2 (9.22g, 12.7mmol), add 800ml of methanol, purge with nitrogen three times, purge with carbon monoxide three times, heat to 60℃ and react for 12h. After the reaction is complete as detected by LC-MS, concentrate the reaction solution under reduced pressure at 40-50℃ to remove the solvent, and purify by column chromatography to obtain compound 16h (32.0g, white solid).

[0215] Step 8: Weigh 16h (32.0g, 109.5mmol), add 384ml of tetrahydrofuran, cool to 0℃ in an ice-water bath, add diisopropylaluminum hydride (1M, 328.5mmol) dropwise, and after the addition is complete, warm to 20-25℃ to react. After the reaction is complete as detected by LC-MS, add 320ml of saturated ammonium chloride solution and 320ml of ethyl acetate dropwise to the reaction system, filter, and concentrate the filtrate under reduced pressure at 40-50℃ to obtain compound 16i (28.6g, yellow solid).

[0216] Step 9: Weigh 16i ​​(28.6 g, 108.2 mmol), add 286 ml of dichloromethane, stir to dissolve, cool to 0°C in an ice-water bath, add Dys-Martin oxidant (55.0 g, 129.8 mmol) in batches, and after the addition is complete, warm to 20-25°C and react for 2 h. After the reaction is complete as detected by TLC, add 300 ml of a 1:1 solution of saturated sodium bicarbonate and saturated sodium thiosulfate and wash twice. Extract the aqueous layer with 300 ml of dichloromethane. Combine the organic layers and concentrate under reduced pressure at 40-50°C. Purify by column chromatography to obtain compound 16j (23.4 g, yellow solid).

[0217] Step 10: Weigh 16J (23.4g, 89.1mmol), add 234ml of tetrahydrofuran, cool to -40℃ in a dry ice bath, add deuterated methyl magnesium iodide (1M, 106.9mmol) dropwise, allow to return to room temperature after addition, stir for 1h, and after the reaction is complete as detected by TLC, add 234ml of saturated ammonium chloride solution, quench with 234ml of water, stir and separate the layers, combine the organic layers and concentrate under reduced pressure at 40-50℃ to obtain compound 16K (25.6g, oil).

[0218] Step 11: Weigh 16k (24.7g, 88.1mmol), add 247ml of dichloromethane, stir until dissolved, cool to 0℃ in an ice-water bath, add Dys-Martin oxidant (44.8g, 105.7mmol) in batches, and after the addition is complete, warm to 20-25℃ and react for 2h. After the reaction is complete as detected by TLC, add 300ml of a saturated sodium bicarbonate: saturated sodium thiosulfate = 1:1 solution to wash twice, extract the aqueous layer with 300ml of dichloromethane, combine the organic layers and concentrate to dryness under reduced pressure at 40-50℃, and purify by column chromatography to obtain compound 16l (23.2g, yellow solid).

[0219] Step 12: Weigh 16L (5.0g, 17.9mmol) and p-toluenesulfonyl hydrazine (3.3g, 17.9mmol), add 100ml of methanol, react at 20-25℃ for 16h, and after the reaction is complete as detected by TLC, filter the reaction solution and dry the filter cake under reduced pressure at 40-50℃ to constant weight to obtain compound 16m (7.20g, white solid).

[0220] Step 13: Weigh 16m (4.50g, 10.0mmol), add 45ml of dichloromethane, cool in an ice-water bath to 0℃, add diisopropylaluminum hydride (1M, 20ml) dropwise, and after the addition is complete, warm to 20-25℃ to react. After the reaction is complete as detected by LC-MS, add 20ml of saturated ammonium chloride solution dropwise to the reaction system, filter, evaporate the filtrate to dryness, and purify by column chromatography to obtain compound 16n (540mg, oily substance).

[0221] Step 14: Weigh 16n (400.0 mg, 1.51 mmol), add 4 ml of dioxane and 4 ml of 48% HBr / H2O, heat to 80℃ and react for 2 h. After the reaction is complete as detected by LC-MS, adjust the pH to 7-8 with saturated sodium carbonate solution, filter, and purify by filter cake column chromatography to obtain compound 16o (200 mg, white solid).

[0222] Step 15: Weigh 16o (100.0 mg, 0.48 mmol), add 3 ml of toluene, add DMF (3.5 mg, 0.048 mmol), cool in an ice-water bath to 0 °C, add thionyl chloride (69.0 g, 0.48 mmol) dropwise, and after the addition is complete, warm to 20-25 °C to react. After the reaction is complete as detected by TLC, concentrate under reduced pressure to remove the solvent, and obtain compound 16p (108 mg, yellow solid).

[0223] Step 16: Add 16p (108.0 mg, 0.48 mmol), int-1 (139.0 mg, 0.48 mmol), DIEA (248.0 mg, 1.92 mmol), and KI (16.0 mg, 0.096 mmol) to 3 ml of acetonitrile. Heat to 80 °C and react for 2 h. After the reaction is complete as detected by LC-MS, cool to room temperature and evaporate to dryness. The crude product is purified by column chromatography to obtain compound 16 (100 mg, white solid). LC-MS: ESI [M+H] + =407.2;1H NMR (400MHz, DMSO-d6) δ11.89(s,1H),8.82–8.71(m,2H),8.48(d,J=1.8Hz,1H),8.10–7.98(m,2H),7.81(s,1H),7.70(d,J=1.8Hz,1H),6. 48(d,J=3.6Hz,1H),3.78(s,2H),3.22(q,J=3.0Hz,2H),2.88(d,J=4.8Hz,3H),2.77(t,J=5.6Hz,2H),2.62(d,J=6.2Hz,2H),2.58(s,2H).

[0224] Example 17: 1'-((7-(ethyl-2,2,2-d3)-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-1',2',3',6'-tetrahydro-[2,4'-bipyridine]-5-nitrile

[0225]

[0226] Step 1: Compound 17a (0.5 g, 2.7 mmol), int-1b (1.02 g, 3.3 mmol), Pd(dppf)Cl2 (0.2 g, 0.27 mmol), and potassium carbonate (0.94 g, 6.8 mmol) were added to a mixed solvent of 7 ml dioxane, 3 ml anhydrous ethanol, and 4 ml water. The mixture was then purged with nitrogen three times and reacted at 90 °C for 2 h under nitrogen protection. After TLC detection of complete reaction, the reaction mixture was cooled to room temperature, and 30 ml dichloromethane and 20 ml water were added. The mixture was separated into layers in a separatory funnel. The aqueous phase was extracted twice with dichloromethane. The combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography to obtain compound 17b (0.7 g, white solid).

[0227] Step 2: Compound 17b (0.5 g, 1.8 mmol) was added to 10 ml of anhydrous methanol, followed by 10 ml of 4 mol / L dioxane hydrochloride solution. The mixture was stirred at room temperature for 0.5-1 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain compound 17c (0.2 g, white solid).

[0228] Step 3: Compound 17c (0.05 g, 0.27 mmol), 16p (0.072 g, 0.32 mmol), N,N-diisopropylethylamine (0.17 g, 1.35 mmol), and potassium iodide (0.22 g, 1.35 mmol) were added to 10 mL of anhydrous acetonitrile. The mixture was stirred at 80 °C for 2 h. After the reaction was complete as monitored by TLC, the reaction solution was concentrated under reduced pressure. The crude product was purified by column chromatography to obtain compound 17 (0.021 g, white solid). LC-MS: ESI [M+H] + =375.2. 1 H NMR (400MHz, DMSO-d6) δ11.85(s,1H),8.96(dd,J=2.2,0.8Hz,1H),8.42(d,J=1.8Hz,1H),8.26(dd,J=8.4,2.3Hz,1H),7.75(d,J=8. 3Hz, 2H), 7.65 (d, J = 1.9Hz, 1H), 6.95 (dd, J = 4.5, 2.6Hz, 1H), 3.74 (s, 2H), 3.22 (d, J = 3.4Hz, 2H), 2.70 (t, J = 5.6Hz, 2H), 2.60 (s, 2H).

[0229] Example 18: 3-(ethyl-2,2,2-d3)-7-((5-fluoro-3',6'-dihydro-[2,4'-bipyridine]-1'(2'H)-yl)methyl)-1,5-naphthidium-2(1H)-one

[0230]

[0231] The preparation of 3-(ethyl-2,2,2-d3)-7-((5-fluoro-3',6'-dihydro-[2,4'-bipyridinyl]-1'(2'H)-yl)methyl)-1,5-naphthidium-2(1H)-one was carried out according to Example 17. LC-MS: ESI [M+H] + =368.2. 1H NMR (400MHz, DMSO-d6) δ11.85(s,1H),8.52(d,J=2.9Hz,1H),8.42(d,J=1.9Hz,1H),7.76(s,1H),7.70(td,J=8.8,3 .0Hz,1H),7.67–7.60(m,2H),6.64(s,1H),3.73(s,2H),3.16(d,J=3.5Hz,2H),2.69(t,J=5.6Hz,2H),2.58(s,2H).

[0232] Example 19: 3-(ethyl-d5)-8-fluoro-7-((5-fluoro-3',6'-dihydro-[2,4'-bipyridin]-1'(2'H)-yl)methyl)quinoxalin-2(1H)-one

[0233]

[0234] Weigh 18c (122.6 mg, 0.49 mmol) into a reaction flask, add 5 mL of DCM, then add TEA to release it, then add glacial acetic acid dropwise to adjust the pH to approximately 5, and finally add 13h (100.0 mg, 0.44 mmol) and stir at room temperature for 2 h. Add sodium triacetoxyborohydride (376.4 mg, 1.78 mmol) to the system, continue stirring for 2 h, then add saturated NH4Cl aqueous solution and stir. Separate the solution, extract the aqueous phase with a 5:1 mixture of DCM and MeOH (30 mL × 3), combine the organic phases and dry them with anhydrous Na2SO4, filter and concentrate to dryness, prepare by HPLC and freeze-dry to give compound 19 (32 mg, 19%) as a white solid. LC-MS: ESI [M+H] + =388.2; 1 H NMR (400MHz, DMSO) δ12.42(s,1H),8.49(d,J=2.8Hz,1H),7.67(td,J=8.7,2.9Hz,1H),7.60(dd,J=8.9,4.5Hz,1H),7.54( d,J=8.3Hz,1H),7.34–7.27(m,1H),6.61(s,1H),3.75(s,2H),3.16(d,J=2.3Hz,2H),2.69(t,J=5.5Hz,2H),2.55(s,2H).

[0235] Example 20: 1'-((2-(ethyl-d5)-5-fluoro-3-oxo-3,4-dihydroquinoxalin-6-yl)methyl)-1',2',3',6'-tetrahydro-[2,4'-bipyridine]-5-nitrile

[0236]

[0237] Weigh 17c (126.1 mg, 0.49 mmol) into a reaction flask, add 5 mL of DCM, then add TEA to release it, then add glacial acetic acid dropwise to adjust the pH to approximately 5, and finally add 13h (100.0 mg, 0.44 mmol) and stir at room temperature for 2 h. Add sodium triacetoxyborohydride (376.4 mg, 1.78 mmol) to the system, continue stirring for 2 h, then add saturated NH4Cl aqueous solution and stir. Separate the solution, extract the aqueous phase with a 5:1 mixture of DCM and MeOH (30 mL × 3), combine the organic phases and dry them with anhydrous Na2SO4, filter, concentrate to dryness, prepare by HPLC, and freeze-dry to give compound 20 (14 mg, 8%) as a white solid. LC-MS: ESI [M+H] + =395.20; 1 H NMR (400MHz, DMSO) δ8.94 (s, 1H), 8.28–8.19 (m, 1H), 7.72 (d, J = 8.4Hz, 1H), 7.55 (d, J = 8.2Hz, 1H), 7.31(t,J=7.6Hz,1H),6.92(s,1H),3.77(s,2H),3.22(s,2H),2.70(d,J=5.2Hz,2H),2.57(s,2H).

[0238] Example 21: 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-2',2',6',6'-d4-6-carboxamide

[0239]

[0240] Step 1: Add compound 21a (6.0 g, 49.2 mmol) to 50 ml of tetrahydrofuran, cool to -78 °C, and add LDA (2 M, 29.5 ml, 59.0 mmol) dropwise while maintaining the temperature between -60 and -78 °C. After the addition is complete, stir for 15 min, then add 40 ml of tetrahydrofuran solution of 21b (19.3 g, 54.1 mmol). After the addition is complete, allow the mixture to return to room temperature naturally, stir for 2 h, and take a sample. After the reaction is complete, add 60 ml of saturated ammonium chloride aqueous solution and stir. Separate the layers, extract the aqueous layer with 100 ml of ethyl acetate, separate the layers, combine the organic layers, wash with 100 ml of saturated brine, separate the layers, concentrate the organic phase under reduced pressure to remove the solvent, and obtain compound 21c (20.0 g, oil).

[0241] Step 2: Add 100 ml of 21c (19.3 g, 28.7 mmol), potassium carbonate (7.93 g, 57.4 mmol), and 21d (8.32 g, 31.6 mmol) to 100 ml of a 1,4-dioxane / water = 10:1 solution. Purge with nitrogen three times, heat to 80 °C and stir for 2 h. After the reaction is complete as detected by LC-MS, cool to room temperature, add 100 ml of water and 100 ml of ethyl acetate, and separate the layers. Extract the aqueous layer three times with 100 ml of ethyl acetate. Combine the organic layers, add anhydrous sodium sulfate and dry, filter, and evaporate to dryness. The crude product obtained is purified by column chromatography to give compound 21e (4.2 g, yellow solid).

[0242] Step 3: Add 21e (4.2 g, 13.0 mmol) to 21 ml of anhydrous methanol, add 40% methylamine aqueous solution (5.1 g, 65.2 mmol), stir at room temperature for 1 h, and after the reaction is complete as detected by LC-MS, concentrate the reaction solution to dryness, add 40 ml of 1,4-dioxane, add dropwise 4M / dioxane hydrochloride solution (16.3 ml, 65.2 mmol), stir at room temperature for 2 h, and after the reaction is complete as detected by LC-MS, filter, and evaporate to dryness to obtain compound 21f (3.4 g, off-white solid).

[0243] Step 4: Compound 21f (3.4 g, 11.5 mmol), DIEA (5.6 g, 46.0 mmol), compound 1 g (2.4 g, 10.5 mmol), and KI (340 mg, 2.1 mmol) were added to 48 ml of acetonitrile. The mixture was heated to 80 °C and stirred for 2 h. After the reaction was confirmed to be complete by LC-MS, 200 ml of saturated sodium bicarbonate solution was added, and the mixture was stirred, filtered, and purified by rotary evaporation column chromatography to obtain compound 21 (2.4 g, white solid). LC-MS: ESI [M+H] + =408.2; 1 H NMR (400MHz, DMSO-d6) δ11.90(s,1H),8.82-8.72(m,2H),8.48(d,J=1.9Hz,1H),8.10-7.99(m,2H),7.82(q,J=1.0Hz,1H) ,7.75-7.66(m,1H),6.48(d,J=1.5Hz,1H),3.78(s,2H),2.88(d,J=4.9Hz,3H),2.64-2.58(m,4H),1.25(t,J=7.4Hz,3H).

[0244] Example 22: 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-2-fluoro-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-2',2',6',6'-d4-6-carboxamide

[0245]

[0246] Step 1: Compound 21c (20 g, 60 mmol), pinacol diboronate (15.2 g, 72 mmol), potassium acetate (11.8 g, 120 mmol), and PdCl2 (dppf) (2.1 g, 3 mmol) were added sequentially to a reaction flask. Under N2 protection, the reaction was carried out at 80 °C overnight. After the reaction was completed, the mixture was filtered, washed with EA, and the filtrate was concentrated. The residue was subjected to column chromatography to obtain compound 22a (15 g, oil).

[0247] Step 2: Add 100 ml of 22a (15 g, 22.3 mmol), potassium carbonate (7.93 g, 57.4 mmol), and 9a (8.32 g, 31.6 mmol) to 100 ml of a 1,4-dioxane / water = 10:1 solution. Purge with nitrogen three times, heat to 80 °C and stir for 2 h. After the reaction is complete as detected by LC-MS, cool to room temperature, add 100 ml of water and 100 ml of ethyl acetate, and separate the layers. Extract the aqueous layer three times with 100 ml of ethyl acetate. Combine the organic layers, add anhydrous sodium sulfate and dry, filter, and evaporate to dryness. The crude product obtained is purified by column chromatography to give compound 22b (12 g, yellow solid).

[0248] Step 3: Add 22b (4.2 g, 13.0 mmol) to 21 ml of anhydrous methanol, add 40% methylamine aqueous solution (5.1 g, 65.2 mmol), stir at room temperature for 1 h, and after the reaction is complete as detected by LC-MS, concentrate the reaction solution to dryness, add 40 ml of 1,4-dioxane, add dropwise 4M / dioxane hydrochloride solution (16.3 ml, 65.2 mmol), stir at room temperature for 2 h, filter after the reaction is complete as detected by LC-MS, and evaporate the filtrate to dryness to obtain compound 22c (3.4 g, off-white solid).

[0249] Step 4: Compound 22c (3.4 g, 11.5 mmol), DIEA (5.6 g, 46.0 mmol), compound 1 g (2.4 g, 10.5 mmol), and KI (340 mg, 2.1 mmol) were added to 48 ml of acetonitrile. The mixture was heated to 80 °C and stirred for 2 h. After the reaction was confirmed to be complete by LC-MS, 200 ml of saturated sodium bicarbonate solution was added, the mixture was stirred and filtered, and purified by rotary evaporation column chromatography to obtain compound 22 (2.4 g, white solid). LC-MS: ESI [M+H] + =426.5; 1H NMR (400MHz, DMSO) δ11.84(s,1H),8.63(d,J=4.8Hz,1H),8.42(t,J=4.4Hz,1H),8.16–8.03(m,1H),7.92(dd,J=7.7,1.5Hz,1H),7.75( s,1H),7.65(s,1H),6.24(s,1H),3.72(s,2H),2.88(t,J=8.1Hz,1H),2.80(d,J=4.8Hz,3H),2.60–2.52(m,3H),1.18(t,J=7.4Hz,3H).

[0250] Example 23: 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N,2-dimethyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-2',2',6',6'-d4-6-carboxamide

[0251]

[0252] Step 1: Add 100 ml of 22a (13 g, 21.2 mmol), potassium carbonate (7.8 g, 55 mmol), and 7a (8.1 g, 28.6 mmol) to 100 ml of a 1,4-dioxane / water = 10:1 solution. Purge with nitrogen three times, heat to 80 °C and stir for 2 h. After the reaction is complete as detected by LC-MS, cool to room temperature, add 100 ml of water and 100 ml of ethyl acetate, and separate the layers. Extract the aqueous layer three times with 100 ml of ethyl acetate. Combine the organic layers, add anhydrous sodium sulfate and dry, filter, and evaporate to dryness. The crude product obtained is purified by column chromatography to give compound 23a (10.6 g, yellow solid).

[0253] Step 2: Add 23a (3.8 g, 11.3 mmol) to 21 ml of anhydrous methanol, add 40% methylamine aqueous solution (4.5 g, 60.5 mmol), stir at room temperature for 1 h, and after the reaction is complete as detected by LC-MS, concentrate the reaction solution to dryness, add 40 ml of 1,4-dioxane, add dropwise 4M / dioxane hydrochloride solution (15 ml, 64 mmol), stir at room temperature for 2 h, and after the reaction is complete as detected by LC-MS, filter, and evaporate the filtrate to dryness to obtain compound 23b (3.2 g, off-white solid).

[0254] Step 3: Compound 23b (3.2 g, 11.2 mmol), DIEA (5.4 g, 45.2 mmol), compound 1 g (2.1 g, 10.5 mmol), and KI (320 mg, 1.9 mmol) were added to 48 ml of acetonitrile. The mixture was heated to 80 °C and stirred for 2 h. After the reaction was confirmed to be complete by LC-MS, 200 ml of saturated sodium bicarbonate solution was added, and the mixture was stirred, filtered, and purified by rotary evaporation column chromatography to obtain compound 23 (1.8 g, white solid). LC-MS: ESI [M+H] + =426.5; 1 H NMR (400MHz, DMSO) δ11.84(s,1H),8.63(d,J=4.8Hz,1H),8.42(t,J=4.4Hz,1H),8.16–8.03(m,1H),7.92(dd,J=7.7,1.5Hz,1H),7.75( s,1H),7.65(s,1H),6.24(s,1H),3.72(s,2H),2.88(t,J=8.1Hz,1H),2.80(d,J=4.8Hz,3H),2.60–2.52(m,3H),1.18(t,J=7.4Hz,3H).

[0255] Bioactivity testing:

[0256] 1. PARP-1 / 2 / 5a enzyme assay

[0257] Experimental materials: PARP1 protein (BPS, Cat. No. 80501), PARP2 protein (BPS, Cat. No. 80502), PARP5A protein (BPS, Cat. No. 80504), Biotin-NAD+ (R&D, Cat. No. 6573), Strep-HRP (Thermo Pierce, Cat. No. 21127), NAD+ (TCI, Cat. No. D0919-5G), quantitative enhanced chemiluminescence HRP substrate kit (Thermo Pierce, Cat. No. 15159), histones (Active Motif, Cat. No. 81167), activated DNA (Genscript, Cat. No. L05182-01&02&03), anti-rabbit IgG, HRP-linked... Antibody (CST, Cat. No. 7074P2), anti-Poly / Mono-ADP Ribose (E6F6A) Rabbit mAb (CST, Cat. No. 83732S), SuperSignal ELISA Femto Substrate (THERMO PIERCE, Cat. No. 37074), and reference compound AZD5305 were purchased from MedChemExpress (MCE).

[0258] 1.1 PARP1 enzyme assay

[0259] 1.1.1 Preparation of buffer solutions: PBST: 1X PBS, 0.05% Tween-20; blocking buffer: 1X PBS, 0.05% Tween-20, 5% BSA; reaction buffer: 50mM Tris-HCl (pH 7.5), 0.005% Tween-20, 0.01% BSA.

[0260] 1.1.2 Coating: Prepare 50 ng / mL Histone coating solution with 1xPBS, transfer 25 μL of the coating solution to a 384-well reaction plate, and coat overnight at 4°C.

[0261] 1.1.3 Washing: After coating, discard the coating solution and wash with PBST solution. The method is to transfer 50 μL of PBST to a 384-well reaction plate, let it stand for 5 minutes, discard the washing solution, refill the plate, repeat the washing 3 times, and finally pat the reaction plate dry and wait for the next step of blocking.

[0262] 1.1.4 Blocking: Transfer 50 μL of blocking solution to a 384-well reaction plate and let stand for 1 hour.

[0263] Washing: After the blocking is completed, discard the blocking solution, wash the plate three times with PBST solution as described in step 2, and finally pat the reaction plate dry.

[0264] 1.1.5 Prepare a 1000-fold dilution of the compound, transfer 1 μL of the compound to a 96-well plate containing 199 μL of reaction buffer, mix well, and then transfer 5 μL of the mixed compound to a 384-well reaction plate.

[0265] 1.1.6 Prepare a 25 / 10-fold PARP1-DNA solution using reaction buffer. Transfer 10 μL of PARP1-DNA solution to a 384-well reaction plate. For the negative control wells, transfer 10 μL of DNA solution. The final concentration of PARP1 is 0.02 nM and the final concentration of DNA is 0.8 nM.

[0266] 1.1.7 Prepare a 25 / 10-fold NAD+ solution using reaction buffer, transfer 10 μL of NAD+ solution to a 384-well reaction plate, with a final NAD+ concentration of 3.5 μM, and incubate at room temperature for 60 minutes.

[0267] 1.1.8 Prepare a 25 / 10-fold NAD+ solution using reaction buffer, transfer 10 μL of NAD+ solution to a 384-well reaction plate, with a final NAD+ concentration of 3.5 μM, and incubate at room temperature for 60 minutes.

[0268] 1.1.9 Washing: After the reaction is complete, discard the reaction solution, wash the plate three times with PBST solution as described in step 2, and finally pat the plate dry.

[0269] 1.1.10 Dilute the primary antibody (anti-Poly / Mono-ADP Ribose Rabbit mAb) 2000 times with blocking buffer, add 20 μL of primary antibody, and incubate at room temperature for 1.5 hours.

[0270] 1.1.11 Washing: Discard the primary antibody, wash three times with PBST solution as described in step 2, and finally pat the reaction plate dry.

[0271] 1.1.12 Dilute the secondary antibody (anti-rabbit IgG, HRP-linked Antibody) 2000 times with blocking buffer, add 20uL of secondary antibody, and incubate at room temperature for 1 hour.

[0272] 1.1.13 Washing: Discard the secondary antibody, wash three times with PBST solution as described in step 2, and finally pat the reaction plate dry.

[0273] 1.1.14 Color development: Mix Femto-ECL Substrate A and Femto-ECL Substrate B in a 1:1 ratio and transfer 25 μL to a 384 reaction plate.

[0274] 1.1.15 Reading: Read the chemiluminescence value RLU using Envision.

[0275] 1.2 PARP2 enzyme assay

[0276] 1.2.1 Preparation of buffer solutions: PBST: 1X PBS, 0.05% Tween-20; blocking buffer: 1X PBS, 0.05% Tween-20, 5% BSA; reaction buffer: 50mM HEPES (pH 7.5), 0.002% Tween-20, 0.1% BSA, 100mM NaCl, 2mM DTT.

[0277] 1.2.2 Coating: Prepare 100 ng / mL Histone coating solution with 1xPBS, transfer 25 μL of the coating solution to a 384-well reaction plate, and coat overnight at 4°C.

[0278] 1.2.3 Washing: After coating, discard the coating solution and wash with PBST solution. The method is to transfer 50 μL of PBST to a 384-well reaction plate, let it stand for 5 minutes, discard the washing solution, refill the plate, repeat the washing 3 times, and finally pat the reaction plate dry and wait for the next step of blocking.

[0279] 1.2.4 Blocking: Transfer 50 μL of blocking solution to a 384-well reaction plate and let stand for 1 hour.

[0280] 1.2.5 Washing: After the blocking is completed, discard the blocking solution, wash the plate three times with PBST solution as described in step 2, and finally pat the reaction plate dry.

[0281] 1.2.6 Prepare a 25 / 10-fold PARP2 solution. Transfer 10 μL of PARP2 solution to a 384-well reaction plate. For the negative control wells, transfer 10 μL of reaction buffer. The final PARP2 concentration is 1.5 nM.

[0282] 1.2.7 Prepare a 2000-fold dilution of the compound. Transfer 50 nL of the compound using an echo converter. Add 19.95 μL of reaction buffer to the compound and mix well. Transfer 5 μL of the mixed compound to a 384-well reaction plate.

[0283] 1.2.8 Prepare a 25 / 10-fold Biotin-NAD+ solution, transfer 10 μL of Biotin-NAD+ solution to a 384-well reaction plate, with a final Biotin-NAD+ concentration of 2 μM, and incubate at room temperature for 60 minutes.

[0284] 1.2.9 Washing: After the reaction is complete, discard the reaction solution, wash the plate three times with PBST solution as described in step 2, and finally pat the reaction plate dry.

[0285] 1.2.10 Prepare a Stre-HRP solution by diluting it with blocking buffer. Transfer 25 μL of Stre-HRP solution to the reaction plate and incubate at room temperature for 1 hour. The final Stre-HRP concentration is 0.1 μg / mL.

[0286] 1.2.11 Washing: Discard the Stre-HRP solution, wash three times with PBST solution as described in step 2, and finally pat the reaction plate dry.

[0287] 1.2.12 Color development: Mix Femto-ECL Substrate A, Femto-ECL Substrate B, and QuantaRed ADHP in a 50:50:1 ratio. Transfer 25 μL of the mixture to a 384 reaction plate and incubate at room temperature for 10 minutes. Add 2.5 μL of QuantaRed StopSolution.

[0288] 1.2.13 Reading: Read the fluorescence value using Paradigm (Ex550 / Em 620).

[0289] 1.3 PARP5A enzyme assay

[0290] 1.3.1 Preparation of buffer solutions: PBST: 1X PBS, 0.05% Tween-20; blocking buffer: 1X PBS, 0.05% Tween-20, 5% BSA; reaction buffer: 50mM HEPES (pH 7.5), 0.002% Tween-20, 0.1% BSA, 100mM NaCl, 2mM DTT.

[0291] 1.3.2 Coating: Prepare 100 ng / mL Histone coating solution with 1xPBS, transfer 25 μL of the coating solution to a 384-well reaction plate, and coat overnight at 4°C.

[0292] 1.3.3 Washing: After coating, discard the coating solution and wash with PBST solution. The method is to transfer 50 μL of PBST to a 384-well reaction plate, let it stand for 5 minutes, discard the washing solution, refill the plate, repeat the washing 3 times, and finally pat the reaction plate dry before the next step of blocking.

[0293] 1.3.4 Blocking: Transfer 50 μL of blocking solution to a 384-well reaction plate and let stand for 1 hour.

[0294] 1.3.5 Washing: After the blocking is completed, discard the blocking solution, wash the plate three times with PBST solution as described in step 2, and finally pat the reaction plate dry.

[0295] 1.3.6 Prepare 25 / 10 times PARP5A solution, transfer 10 μL of PARP5A solution to a 384-well reaction plate, and transfer 10 μL of reaction buffer to the negative control wells. The final concentration of PARP5A is 10 nM.

[0296] 1.3.7 Prepare a 2000-fold dilution of the compound. Transfer 50 nL of the compound using an echo converter. Add 19.95 μL of reaction buffer to the compound and mix well. Transfer 5 μL of the mixed compound to a 384-well reaction plate.

[0297] 1.3.8 Prepare a 25 / 10-fold Biotin-NAD+ solution, transfer 10 μL of Biotin-NAD+ solution to a 384-well reaction plate, with a final Biotin-NAD+ concentration of 2 μM, and incubate at room temperature for 60 minutes.

[0298] 1.3.9 Washing: After the reaction is complete, discard the reaction solution, wash the plate three times with PBST solution as described in step 2, and finally pat the plate dry.

[0299] 1.3.10 Prepare a Stre-HRP solution by diluting with blocking buffer, transfer 25 μL Stre-HRP solution to the reaction plate, and incubate at room temperature for 1 hour. The final Stre-HRP concentration is 0.1 μg / mL.

[0300] 1.3.11 Washing: Discard the Stre-HRP solution, wash three times with PBST solution as described in step 2, and finally pat the reaction plate dry.

[0301] 1.3.12 Color development: Mix Femto-ECL Substrate A, Femto-ECL Substrate B, and QuantaRed ADHP in a 50:50:1 ratio. Transfer 25 μL of the mixture to a 384 reaction plate and incubate at room temperature for 10 minutes. Add 2.5 μL of QuantaRed StopSolution.

[0302] 1.3.13 Reading: Read the fluorescence value using Paradigm (Ex550 / Em 620).

[0303] The test results are shown in Table 2 below:

[0304] Table 2. PARP-1 / 2 / 5a enzyme test results

[0305]

[0306]

[0307] Conclusion: The compounds of this invention have a significant inhibitory effect on PARP1, but a weak inhibitory effect on PARP2 / 5a, indicating that the compounds of this invention selectively inhibit PARP-1.

[0308] 2. Cell anti-proliferation activity test:

[0309] BRCA mutant MDA-MB-436 cells were cultured in DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin, and incubated at 37°C in a 5% CO2 saturated incubator. When cells reached 80% confluence, they were harvested, centrifuged at 300g for 10 min, and seeded at 1200 cells / well in 96-well plates. After 24 h, different final concentrations of PARPi (0, 0.01, 0.1, 1, 10, 100, 1000 nM) were added, and the cells were cultured for another 72 h. The medium was then changed (with the same final concentration of PARPi added), and the cells were cultured for another 96 h. The OD values ​​at 450 nM were measured using the CCK8 assay, and the cell inhibition rate was calculated as follows: Inhibition rate % = 1 - (mean OD value of the treated group - mean OD value of the Blank group) / (mean OD value of the control group - mean OD value of the Blank group) * 100%.

[0310] BRCA wild-type DLD-1 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin, and incubated at 37°C in a 5% CO2 saturated incubator. When cells reached 80% confluence, they were harvested, centrifuged at 300g for 10 min, and seeded at 1000 cells / well in 96-well plates. After 24 h, different final concentrations of PARPi (0, 1, and 10 μM) were added, and the cells were cultured for another 72 h. The medium was then changed (with the same final concentration of PARPi added), and the cells were cultured for another 96 h. The OD values ​​at 450 nM were measured using the CCK8 assay, and the cell inhibition rate was calculated.

[0311] Inhibition rate % = 1 - (mean OD value of the treatment group - mean OD value of the Blank group) / (mean OD value of the control group - mean OD value of the Blank group) * 100%.

[0312] The test results are shown in Table 3 below:

[0313] Table 3 Results of cell antiproliferation activity assay

[0314]

[0315]

[0316] Conclusion: The compound of the present invention has a significant inhibitory effect on BRCA-mutated MDA-MB-436 cells and no obvious inhibitory effect on BRCA wild-type DLD-1 cells, indicating that the compound of the present invention specifically inhibits tumor cells with homologous recombination deficiency.

[0317] 3. Pharmacokinetic Evaluation of the Compound in Balb / c Mice

[0318] Experimental Purpose: To understand the pharmacokinetics of the compound.

[0319] Experimental Basis: Technical Guidelines for Non-Clinical Pharmacokinetic Studies of Chemical Drugs, 2014.

[0320] Experimental Scheme: By intravenous administration (1 mg·kg-1) and gavage administration (1 mg·kg-1) to Balb / c mice, the pharmacokinetics of the compound was investigated.

[0321] Sample Preparation: Weigh about 0.2 mg of the compound, dissolve it with 10 μL DMSO, and then add sodium chloride injection solution to prepare a 0.1 mg·mL-1 compound solution for use in administration.

[0322] Sample Collection: Six Balb / c mice (Chengdu Dashuo Experimental Animal Co., Ltd., License No.: SCXK(Sichuan)2020-030), male. Three mice were administered intravenously (IV) at 1 mg·kg-1 and three mice were administered by gavage (PO) at 1 mg·kg-1. Approximately 0.05 mL of blood was collected at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, 24 h, and 48 h after administration. The collected blood was centrifuged at 3500 rpm for 15 min, and the supernatant plasma was collected and stored at -40 °C for future testing. The blood drug concentration was quantitatively analyzed by LC-MS / MS analysis method, and pharmacokinetic parameters such as peak time (Cmax), area under the concentration-time curve (AUC(0-t)), half-life (T 1 / 2 ), clearance rate (CL), volume of distribution at steady state (Vdss), bioavailability (F), etc. were calculated.

[0323] The results of pharmacokinetic evaluation are shown in Table 4 below:

[0324] Table 4 Pharmacokinetic Test Results of the Compound in Balb / c Mice

[0325]

[0326]

[0327] Conclusion: The compounds of this invention exhibit favorable pharmacokinetic properties in Balb / c mice, including good oral bioavailability, exposure, half-life, and clearance. After oral gavage administration of 1 mg / kg, the C6 concentration of compound 6... max It is superior to reference compounds 5305, 1, 2, 3, 15 and 21.

[0328] 4. Pharmacokinetic evaluation of the compound in SD rats

[0329] Experimental objective: To understand the pharmacokinetics of the compound.

[0330] Experimental basis: Technical Guidelines for Non-Clinical Pharmacokinetic Studies of Chemical Drugs, 2014.

[0331] Experimental protocol: The pharmacokinetics of the compound were investigated by oral and intravenous administration to SD rats.

[0332] Experimental Procedure: Weigh the compound, add a small amount of DMSO, then add sodium chloride solution for injection to prepare a solution for drug administration. Six male SD rats were administered the drug orally. Approximately 0.1 mL of blood was collected at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, and 24 h after administration. The blood samples were centrifuged at 3500 rpm for 15 min, and the supernatant plasma was collected. 5 μL of plasma was placed in an EP tube, and 100 μL of acetonitrile containing 20 ng·ml⁻¹ internal standard SAHA was added to precipitate the protein. The tube was vortexed for 30 s and centrifuged at 13000 rpm for 15 min. The supernatant was collected and placed in a sample vial for analysis. Standard curve range: 10–10000 ng·ml⁻¹.

[0333] The pharmacokinetic evaluation results are shown in Table 5 below:

[0334] Table 5. Pharmacokinetic results of the compounds in SD rats.

[0335]

[0336] Conclusion: The compounds of this invention exhibit favorable pharmacokinetic properties in SD rats, including good oral bioavailability, exposure, half-life, and clearance. Compound 6 has a C0.05 max It is superior to reference compounds AZD5305, 1 and 21.

[0337] 5. In vivo pharmacodynamic studies of compounds 1 and 6 on a subcutaneous xenograft tumor model of breast cancer MDA-MB-436 in nude mice.

[0338] (1) Major instruments and equipment

[0339] CO2 cell incubator: Yamato brand, model IP610; Biosafety cabinet: Suzhou Antai brand, model BSC-1304ⅡA2; Room temperature centrifuge: Thermo Scientific brand, model SORVALLST 16;

[0340] Digital inverted microscope: Olympus brand, model CKX3-SLP; Constant temperature water bath: Shanghai Yuejin Medical Instrument Co., Ltd., model HSW-420; Cell counting chamber: Shanghai Qiujing Biochemical Reagent Instrument Co., Ltd., model XB.K.25; Liquid nitrogen tank: Thermo brand, model CY50935-70; Refrigerator: Qingdao Haier brand, model BCD-601WDGX; Medical low temperature freezer: Thermo brand, model ULTS1651 (USA);

[0341] Pure water system: Millipore (USA), model F7PNO9748; Vertical autoclave: Yamato, model DKN812C; 1mL disposable sterile syringe: Shanghai Kangdelai Enterprise Development Group Co., Ltd.; 1mL disposable sterile insulin syringe: BD brand; Weight scale: Shanghai Hengping Instrument Factory, model JY2002; Analytical scale: SARTORIUS, model BCE95I-1CEU;

[0342] Thermometer and hygrometer: Wuqiang Thermometer and Hygrometer Manufacturing Center, Hebei Province, No. 30260102; Mini centrifuge: Thermo brand, model SORVALL LEGEND MICRO 17Centrifuge; Mini mixer: Yeasen brand, model ES-VM25; Metal bath: SCILOGEX brand, model SCL120-S.

[0343] (2) Main software and data processing systems

[0344] Graphpad Prism, version 6.0, from Graphpad Software Ltd., is a software used for organizing and quantifying data to create bar charts.

[0345] (3) Laboratory animals

[0346] Strain: NOD / SCID, Grade: SPF, Source: Beijing Vital River Biotechnology Co., Ltd.

[0347] (4) Modeling methods

[0348] Tumor cell culture and preparation: Routine passage and culture of tumor cell lines were performed using a culture medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin. When cells reached 80% confluence, they were collected and centrifuged at 300g for 10 min. Cells were washed three times with pre-chilled PBS, centrifuged again at 300g for 10 min, resuspended in PBS, and counted using a hemocytometer. Cell concentration was adjusted. After collection, the cell suspension was pre-chilled on ice. 100 μL of the cell suspension was subcutaneously injected into the dorsal axilla of mice. Inoculation was performed when the tumor volume reached 200-300 mm². 3 Treatment will begin in groups at that time.

[0349] (5) Observation and detection indicators

[0350] General observation: Animals to be observed: All surviving laboratory animals to be observed; Observation time: Twice a day; Observation content: Including but not limited to general appearance, behavioral status, toxic symptoms in the eyes, mouth, nose and mouth, ears, hair, feces, urine, genitals, etc. If any abnormalities occur, they must be described in detail.

[0351] (6)Weight

[0352] Animals tested: All surviving experimental animals planned for testing; Testing time: The weight of all animals was measured before modeling for experimental grouping. After modeling, the weight of mice was measured every 2 days, and the weight changes of mice were recorded in real time. Validity criteria: Tumor volume (TV), relative tumor volume (RTV), and relative tumor proliferation rate in the subcutaneous xenograft model. The long and short diameters of the tumor were measured every 2 days using calipers to dynamically observe the antitumor effect of the test drug. The formula for calculating tumor volume (TV) is: TV (mm3) = a × b² × 0.5; where a and b represent the long and short diameters, respectively.

[0353] The relative tumor volume (RTV) is calculated based on the measurement results using the formula: RTV = Vt / V0, where V0 is the total tumor volume (TV) measured at the time of administration (d0) for each group, and Vt is the TV measured at each subsequent measurement for that group. The evaluation index for antitumor activity is the relative tumor proliferation rate (T / C%), calculated using the following formula:

[0354] T / C(%)=TRTV / CRTV×100%;

[0355] TRTV: relative tumor volume in the treatment group; CRTV: relative tumor volume in the negative control group. Efficacy evaluation criteria: T / C (%) > 60 was considered ineffective; T / C (%) ≤ 60, and statistical analysis showed P < 0.05, was considered effective. Tumor weight measurement and tumor inhibition rate (%) calculation: At the end of treatment, animals were sacrificed, the tumor was dissected, weighed, and photographed. The tumor inhibition rate (%) was calculated using the following formula:

[0356] Tumor inhibition rate (tumor growth inhibition rate, %) = (average tumor weight of negative control group (g) - average tumor weight of treatment group (g)) / average tumor weight of negative control group (g) × 100%;

[0357] Validity criteria: A tumor inhibition rate (i.e., tumor growth inhibition rate) < 40% is considered invalid; a tumor inhibition rate ≥ 40% with statistically significant P < 0.05 is considered valid. Comprehensive criteria for subcutaneous tumor models: A model is considered valid if either of the above two validity criteria (relative tumor proliferation rate and tumor inhibition rate) is met.

[0358] (7) Data collection and analysis

[0359] All raw data within the facility were collected manually or using a data acquisition system according to the experimental protocol and relevant regulations of the research institution. Manually collected data could be transcribed into software such as Excel for analysis and reporting. All data were quantitative data. Animal experimental data in the tables are expressed as mean ± standard deviation (Mean ± SD), and experimental data in the images are described as mean ± standard error (Mean ± SEM). t-tests and survival analyses were performed using Excel and GraphPad Prism software.

[0360] Experimental Procedure: Forty-two female NOD / SCID mice were randomly divided into seven groups: a blank control group, compound 6 (0.03, 0.1, 0.3 mg / kg) dose groups, reference compound AZD5305 0.1 mg / kg, compound 1 0.1 mg / kg, and olaparib 100 mg / kg groups, with six mice in each group. All mice were administered the medication orally once daily. Body weight was measured every two days, and tumor length and width were measured using calipers. After 24 days of administration, the tumor-bearing mice were anesthetized and euthanized. Tumor tissue was dissected, weighed, and photographed to calculate the tumor inhibition rate.

[0361] Conclusion: The experimental results are as follows Figure 1As shown in Table 6, after cell seeding, each treatment group was administered the drug for 24 consecutive days. Compared with the blank control group, each treatment group (AZD5305 0.1 mg / kg, compound 1 0.1 mg / kg, Olaparib 100 mg / kg, compound 6 0.03, 0.1, and 0.3 mg / kg) significantly inhibited tumor growth in the NOD / SCID mouse subcutaneous tumor model of human breast cancer cell line MM436. The tumor inhibition rates of each treatment group were 80.79%, 82.18%, 62.19%, 49.77%, 86.73%, and 95.83%, respectively; the relative tumor proliferation rates (T / C%) were 15.2%, 17.0%, 33.4%, 40.8%, 12.6%, and 3.8%, respectively. Among all treatment groups, the compound 6 0.3 mg / kg group showed the best tumor inhibition effect, with complete tumor regression in 2 mice at the treatment endpoint (2 / 6). The compound 6 0.1 mg / kg group showed better tumor inhibition than the reference compound AZD5305 and compound 1 at the same dose, and was far superior to the reference drug Olaparib 100 mg / kg group (P<0.01). At the same time, there were significant dose-response differences among the low, medium and high dose groups of compound 6 (P<0.001, P<0.01).

[0362] Table 6. Tumor inhibition rates of compounds 1 and 6 in MDA-MB-436 nude mouse models after 24 days.

[0363]

[0364] VS Blank Group: c, P <0.001

[0365] 6. Challenge experiments of compounds 1, 6, and 21 on a subcutaneous xenograft tumor model of breast cancer MDA-MB-436 in nude mice.

[0366] Experimental Procedure: Fifteen female NOD / SCID mice were randomly divided into three groups: compound 1 (1 mg / kg), compound 6 (1 mg / kg), and compound 21 (1 mg / kg), with five mice in each group. All mice were administered the compound orally once daily. The mice were weighed every two days, and the length and width of the tumors were measured using calipers.

[0367] Conclusion: The experimental results are as follows Figure 2 As shown, compounds 1, 6, and 21 of this invention still produce good anti-tumor efficacy against large-volume tumor models, leading to complete tumor regression. Compound 6 exhibits faster tumor regression than compounds 1 and 21, indicating that at the same dosage, compound 6 has a stronger in vivo anti-tumor efficacy compared to compounds 1 and 21.

[0368] 7. In vivo pharmacodynamic study of compound 6 in a mouse model of PDX gastric cancer

[0369] Procedure: A stable passaged tumor-bearing mouse model was constructed using tissue from gastric cancer patients. In this study, tumor fragments from mice that had been stably passaged three times were subcutaneously inoculated. The experiment was divided into 8 groups, with 6 mice in each group. The negative control group (Control) received physiological saline, 0.2 ml / mouse / dose, orally once a day; the carboplatin control group (Car) received 20 mg / kg, 0.2 ml / mouse / dose, intraperitoneally once every 7 days. The test drug compound 6 was administered orally at doses of 0.3, 1, and 3 mg / kg, 0.2 ml / mouse / dose, once a day; the combination group received compound 6 plus carboplatin.

[0370] The specific dosing regimen and grouping are shown in Table 7.

[0371] Table 7 Experimental Grouping Table

[0372]

[0373] Conclusion: The experimental results are as follows Figure 3 As shown, different doses of compound 6, when combined with carboplatin, exhibited dose-dependent inhibitory effects on a gastric cancer PDX model, especially the 3 mg / kg dose group, where tumor growth was significantly inhibited when compound 6 was combined with carboplatin.

[0374] 8. In vivo pharmacodynamic study of compound 6 on a subcutaneous xenograft tumor model of breast cancer in nude mice (SUM14PT model).

[0375] Procedure: This study used subcutaneous inoculation of SUM14PT tumor cells to establish a mouse model. The experiment was divided into 8 groups, with 6 mice in each group. The negative control group (Control) received saline, 0.2 ml / mouse / dose, orally once daily. The carboplatin control group (Car) received 20 mg / kg, 0.2 ml / mouse / dose, intraperitoneally once every 7 days. The olaparib control group received 100 mg / kg, 0.2 ml / mouse / dose, orally once daily. The test drug compound 6 received 0.3 and 1 mg / kg, 0.2 ml / mouse / dose, orally once daily. The combination groups received either olaparib or compound 6 + carboplatin.

[0376] The specific dosing regimen and grouping are shown in Table 8.

[0377] Table 8 Experimental Grouping Table

[0378]

[0379] Conclusion: The experimental results are as follows Figure 4As shown. Compound 6 at 0.3 and 1 mg / kg, administered orally once daily, showed better antitumor effects than olaparib at 100 mg / kg, administered orally once daily; Compound 6 at 1 mg / kg, administered orally once daily, combined with carboplatin, showed better efficacy than olaparib at 100 mg / kg, administered orally once daily, combined with carboplatin.

Claims

1. A compound of the formula or a pharmaceutically acceptable salt thereof, characterized in that: The formula is The structure is as follows: ； in: represents a double bond; R1is selected from C 1-4 deuteroalkyl; X1is selected from N or C(R 5a ), X2is selected from N or C(R 5b ), X3is selected from C(R 5c ), X1and X2have and only have 1 selected from N; R 2a and R 2b is independently selected from hydrogen; R3 is selected from deuterium, fluorine, and C. 1-4 Alkyl or C 1-4 At least one of the deuterated alkyl groups; R 3a absent; R4 is selected from -CONHR7; R 5a , R 5b , and R 5c are independently selected from hydrogen; R6 is selected from hydrogen, fluorine, or chlorine; R7is selected from C 1-4 alkyl or C 1-4 deuteroalkyl; X5is selected from C(R 9a ), X6is selected from C(R 9b ), X7is selected from C(R 9c ), and X8is selected from nitrogen; R 9a , R 9b , R 9c is independently selected from hydrogen, fluoro, chloro, C 1-4 alkyl or C 1-4 deuteroalkyl; n1 is 0; n3 is selected from 0.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: R1 is selected from deuterated methyl or deuterated ethyl.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: R4 is selected from methylaminoyl or deuterated methylaminoyl.

4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: It has the structure described in Formula VI-1: ； in: R6 is selected from hydrogen or fluorine; R 9a selected from hydrogen, fluorine, chlorine, C 1-4 alkyl or C 1-4 deuteroalkyl; n1 is 0; R1, R 2a , R 2b , R3 and R7 are as described in claim 1.

5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: It has the structure described in Formula VI-3: ； in: R6 is selected from hydrogen or fluorine; R 9a Selected from hydrogen, fluorine, chlorine, C 1-4 Alkyl or C 1-4 Deuterated alkyl groups; n1 is 0; R1, R 2a , R 2b , R3 and R7 are as described in claim 1.

6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: The compound is selected from: , , , , or .

7. A pharmaceutical composition characterized by: It uses the compound or a pharmaceutically acceptable salt of any one of claims 1 to 6 as the active ingredient, supplemented by a pharmaceutically acceptable carrier.

8. Use of the compound of any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 7 in the preparation of a medicament for the prevention and / or treatment of PARP1 enzyme-related diseases.

9. Use according to claim 8, characterized in that: The PARP1 enzyme-related disease is a tumor.

10. Use according to claim 9, characterized in that: The tumor contains one or more cancer cells that have a BRCA1 or BRCA2 defective phenotype.

11. Use according to claim 9, characterized in that: The tumors mentioned are breast cancer, ovarian cancer, primary peritoneal cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer, glioblastoma, or lung cancer.

Images