Heterocyclic compound and application thereof

Abstract

The invention discloses a heterocyclic compound and application thereof, and belongs to the technical field of chemical medicines. The heterocyclic compound shown in the formula (II) provided by the invention can be used as a PARP1 inhibitor, has the advantages of high activity and high selectivity, and also has excellent pharmacokinetic properties and excellent safety. # imgabs0 #

Description

Heterocyclic compounds and uses thereof Technical Field

[0001] The present invention belongs to the field of chemical medicine and relates to a class of heterocyclic compounds and uses thereof. Background Art

[0002] As cells grow, their DNA is constantly damaged by various internal and external factors. The most severe types of DNA damage 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, cancer, and even direct cell death. The repair of single-strand breaks in DNA primarily relies on the enzyme PARP. Double-strand breaks can be repaired through two methods: non-homologous end joining repair and homologous recombination repair. Homologous recombination repair is a high-fidelity, error-free repair method and the primary pathway for double-stranded DNA repair. Numerous proteins are involved in homologous recombination repair, the most well-known of which are the 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 are selectively inhibited by PARP inhibitors. Based on this research result, scholars proposed the concept of synthetic lethality: the loss of either BRCA or PARP gene is not lethal in itself, but the simultaneous inactivation of both will lead 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 shown excellent clinical efficacy in patients with homologous recombination-deficient cancers. However, whether used as a single agent or in combination therapy, hematological toxicity (anemia, neutropenia, and thrombocytopenia) and other toxicities limit the application of this type of drug. Related studies have shown (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-benzenesulfonamide (Pazopanib), a novel and potent vascular endothelial growth factor receptor inhibitor. [J]. Journal of Medicinal Chemistry, 2008, 51(15): 4632.) These adverse reactions may be due to the inhibition of PARP2 by marketed PARP inhibitors, which is not essential for efficacy. Highly selective PARP1 inhibitors can reduce hematological toxicity, increase the therapeutic safety window, and increase the potential for combination with other chemotherapy or targeted drugs.

[0004] Therefore, there is an unmet clinical need for effective and safe PARP inhibitors, particularly PARP inhibitors that are selective for PARP1. The novel PARP1 inhibitors described in the present invention are unexpectedly selective for PARP1 over other PARP family members (such as PARP2, PARP3, PARP5a, and PARP6) and can be used to treat diseases related to PARP function.

[0005] Summary of the Invention

[0006] The purpose of the present invention is to provide a class of heterocyclic compounds and uses thereof to achieve highly selective and efficient prevention or treatment of diseases related to PARP function.

[0007] In a first aspect, the present invention provides a compound represented by Formula II or a pharmaceutically acceptable form thereof, wherein the structure of Formula II is as follows:

[0008] in:

[0009] Indicates a single bond or a double bond;

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

[0011] X1 is selected from N or C(R5a ), X2 is selected from N or C(R 5b ), X3 is selected from N or C(R 5c ), X1, X2 and X3 have only one selected from N;

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

[0013] R3 is selected from deuterium, fluorine, C 1-4 Alkyl or C 1-4 at least one of a deuterated alkyl group;

[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, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluoroalkyl or -CONHR7;

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

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

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

[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 Fluorinated alkyl, C 1-4 Alkoxy or C 1-4 Fluoroalkoxy;

[0021] n1 is an integer from 0 to 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, N-oxides, isotopically labeled substances, metabolites, or prodrugs.

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

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

[0026] In some embodiments of the present invention, in the compound represented by formula II or a pharmaceutically acceptable form thereof, 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 the above formula II or a pharmaceutically acceptable form thereof, R3 is selected from deuterium or methyl.

[0028] In some embodiments of the present invention, the compound represented by the above formula II or a pharmaceutically acceptable form thereof, When it is a double bond, R 3a 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 of formula II or a pharmaceutically acceptable form thereof, R4 is selected from fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluorinated alkyl, C 1-4 Alkyl, C 1-4 Deuterated alkylaminoacyl, C 1-4 Fluorinated alkylaminoacyl or 3-6 membered cycloalkylaminoacyl.

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

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

[0032] In some embodiments of the present invention, in the compound represented by the above formula II or a pharmaceutically acceptable form thereof, R 5a 、R 5b and R 5c are independently selected from hydrogen, fluorine, chlorine or methyl.

[0033] In some embodiments of the present invention, based on the compound represented by the above formula II or a pharmaceutically acceptable form thereof, the present invention further provides a compound having the structure represented by formula VI-1:

[0034] in:

[0035] R6 is selected from hydrogen or fluorine;

[0036] R 9a Selected from hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Fluorinated alkyl;

[0037] n1 is an integer from 0 to 6.

[0038] In some embodiments of the present invention, based on the compound represented by the above formula II or a pharmaceutically acceptable form thereof, the present invention further provides a compound having the structure represented by formula VI-3:

[0039] in:

[0040] R6 is selected from hydrogen or fluorine;

[0041] R 9a Selected from hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Fluorinated alkyl;

[0042] n1 is an integer from 0 to 6.

[0043] In some more preferred embodiments of the present invention, the following compounds and pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope labels, metabolites or prodrugs thereof are provided:

[0044] In some more preferred embodiments of the present invention, the following compounds and pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope labels, metabolites or prodrugs thereof are provided:

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

[0046] In a second aspect, the present invention provides a pharmaceutical composition, which uses 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 an active ingredient, supplemented by a pharmaceutically acceptable carrier.

[0047] A further object of the present invention is to provide a method for preparing the pharmaceutical composition of the present invention, which comprises 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.

[0048] The pharmaceutically acceptable carrier that can be used in the pharmaceutical composition of the present invention is a pharmaceutically acceptable carrier. Examples of suitable pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (2005).

[0049] The pharmaceutical composition can be administered in any form, as long as it prevents, alleviates, prevents or cures the symptoms of a human or animal patient. For example, it can be prepared into various suitable dosage forms according to the route of administration.

[0050] In other embodiments, the administration of the compound or pharmaceutical composition of the present invention can be combined with another treatment method. The other treatment method can be selected from, but not limited to: radiation therapy, chemotherapy, immunotherapy, or a combination thereof.

[0051] The present 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 a pharmaceutical composition of the present invention. In some embodiments, the formulation is in the form of a solid formulation, a semisolid formulation, a liquid formulation, or a gaseous formulation.

[0052] A further object of the present invention is to provide an article of manufacture, for example, in the form of a kit. Articles of manufacture as used herein are intended to include, but are not limited to, kits and packaging. The article of manufacture of the present invention comprises: (a) a first container; (b) a pharmaceutical composition located in the first container, wherein the composition comprises: a first therapeutic agent, the 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) an optional package insert indicating that the pharmaceutical composition can be used to treat a neoplastic condition (as defined below); and (d) a second container.

[0053] The first container is a container for holding a pharmaceutical composition. This container can be used for 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 preparing, holding, storing, or dispensing pharmaceutical products.

[0054] The second container is a container for accommodating the first container and optional package insert. Examples of the second container include, but are not limited to, boxes (e.g., paper or plastic boxes), boxes, cartons, bags (e.g., paper or plastic bags), pouches, and sacks. The package insert can be physically adhered to the outside of the first container via a cable tie, glue, staples, or other adhesion methods, or it can be placed inside the second container without any physical tool for adhering to the first container. Alternatively, the package insert is located outside the second container. When located outside the second container, it is preferred that the package insert is physically adhered via a cable tie, glue, staples, or other adhesion methods. Alternatively, it can abut or contact the outside of the second container without physical adhesion.

[0055] The package insert is a trademark, label, or indicia that lists information about the pharmaceutical composition within the first container. The information listed is typically determined by the regulatory agency (e.g., the U.S. Food and Drug Administration) that governs the region in which the product is to be sold. Preferably, the package insert specifically lists the indications for which the pharmaceutical composition is approved. The package insert can be made of any material from which the information contained therein or thereon can be read. Preferably, the package insert is a printable material (e.g., paper, plastic, cardboard, foil, adhesive paper, or plastic, etc.) onto which the desired information can be formed (e.g., printed or applied).

[0056] In a third aspect, 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 pharmaceutically acceptable forms thereof, or the pharmaceutical compositions of the present invention in the preparation of drugs for preventing or treating PARP1 enzyme-related diseases.

[0057] The present invention provides a method for preventing or treating PARP1 enzyme-related diseases, comprising administering 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 to an individual in need thereof.

[0058] The present invention provides 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 for use in preventing or treating PARP1 enzyme-related diseases.

[0059] The present invention provides a method for preventing or treating PARP1 enzyme-related diseases in combination with 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, wherein the additional treatment method includes but is not limited to: radiotherapy, chemotherapy, immunotherapy, or a combination thereof.

[0060] In some embodiments, the PARP1 enzyme-related disease is a disease that is sensitive or responsive to PARP1 enzyme inhibition.

[0061] In some embodiments, the PARP1 enzyme-related disease is a tumor-related disorder.

[0062] In some preferred embodiments, the oncological disorder is deficient in a HR-dependent DNA DSB repair pathway.

[0063] In some preferred embodiments, the neoplastic disorder comprises one or more cancer cells that have a reduced or absent ability to repair DNA DSBs by HR relative to normal cells.

[0064] In some preferred embodiments, the cancer cells have a BRCA1 or BRCA2 deficient phenotype.

[0065] In some embodiments, the PARP1 enzyme-related disease is a tumor-related disorder, including but not limited to solid and hematological malignancies. In further embodiments, the tumor-related disorder includes but is 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 tissue 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 lymphocytic leukemia (ALL) and chronic myeloid leukemia (CML)), multiple myeloma or lymphoma.

[0066] In some preferred embodiments, the PARP1 enzyme-related disease is breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer or lung cancer.

[0067] In a further preferred embodiment, the compounds of the present invention can be used in combination with chemoradiotherapy or immunotherapy to prevent or treat cancer.

[0068] Beneficial effects of the present invention:

[0069] The present invention provides a novel class of highly active and highly selective PARP1 inhibitors capable of achieving 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 and weak inhibitory effect on non-homologous recombination-deficient cells; (4) excellent pharmacokinetic properties (e.g., good bioavailability, suitable half-life and duration of action); (5) excellent safety (lower toxicity and / or fewer side effects, wider therapeutic window), etc.

[0070] Definition of terms:

[0071] 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 "include," "comprising," "having," "containing," or "involving," and their variations herein, are inclusive or open-ended and do not exclude other unrecited elements or method steps. It should be understood by those skilled in the art that the above terms, such as "comprising," encompass the meaning of "consisting of."

[0072] In the present invention, "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 to mean that the composition includes "one or more" pharmaceutically acceptable excipients.

[0073] For example, the statement "C 1-4 " should be understood to include any sub-ranges therein and each point value, such as C 2-4 、C 3-4 、C 1-2 、C 1-3 、C 1-4 etc., as well as C1, C2, C3, C4, etc.

[0074] As used herein, unless otherwise stated, Indicates a single bond or a double bond.

[0075] In the present invention, unless otherwise specified, halogen means fluorine, chlorine, bromine or iodine.

[0076] In the present invention, unless otherwise specified, "alkyl" includes linear or branched monovalent saturated hydrocarbon groups. For example, alkyl includes 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, the C1-4 in "C1-4 alkyl" refers to a group containing 1, 2, 3, or 4 carbon atoms in a linear or branched form.

[0077] In the present invention, unless otherwise specified, "cycloalkyl", "carbocycle" 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 spiro rings, such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[5.2.0]nonyl, decalinyl, etc. For example, "C 3-12 cycloalkyl" refers to a cycloalkyl group having 3-12 ring carbon atoms (such as 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12). The cycloalkyl or cycloalkylene group in the present invention is optionally substituted with one or more substituents described herein.

[0078] In the present invention, unless otherwise specified, "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 group optionally substituted by one or more (e.g. 1-3) fluorine atoms. 1-4 Alkyl. It will be understood by those skilled in the art that when there are more than one fluorine atom substituent, the fluorine atoms may be the same or different and may be located on the same or different C atoms. Examples of haloalkyl groups include -CH2F, -CHF2, -CF3, -C2F5, -CH2CF3, -CH2CH2CF3, etc. The fluoroalkyl groups of the present invention are optionally substituted with one or more substituents described herein.

[0079] The present invention also includes all pharmaceutically acceptable isotopically labeled compounds, which are identical to the compounds of the present invention except that one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature. Examples of isotopes suitable for inclusion in the compounds of the present invention include, but are not limited to, isotopes of hydrogen (e.g., deuterium ( 2 H), tritium ( 3H)); carbon isotopes (e.g. 13 C and 14 C); isotopes of chlorine (such as 37Cl); isotopes of iodine (such as 125 I); isotopes of nitrogen (e.g. 13 N and 15 N); oxygen isotopes (e.g. 17 O and 18 O); isotopes of phosphorus (such as 32 P); and sulfur isotopes (e.g. 34 S).

[0080] As used herein, "polymorph" refers to different solid crystalline phases of certain compounds of the present invention resulting from the presence of two or more different molecular arrangements in the solid state. Certain compounds of the present invention may exist in more than one crystalline form, and the present invention is intended to encompass various crystalline forms and mixtures thereof. Typically, crystallization produces solvates of the compounds of the present invention. The term "solvate," as used herein, refers to an aggregate comprising one or more molecules of the compound of the present 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. Thus, the compounds of the present invention may exist as hydrates, including monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, and the like, as well as corresponding solvated forms. The compounds of the present invention may form true solvates, but in some cases, they may only retain adventitious water or a mixture of water and a portion of adventitious solvent. The compounds of the present invention may react in a solvent or precipitate or crystallize from a solvent. Solvates of the compounds of the present invention are also encompassed by the present invention. The present invention also encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be single polymorphs or mixtures of more than one polymorph in any ratio.

[0081] In the present invention, "stereoisomer" means an isomer formed due to at least one asymmetric center. In compounds with one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers can be produced. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of the present 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-ketone tautomers, nitroso-oxime tautomers, and imine-enamine tautomers. It is to be understood that the scope of the present invention encompasses all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0082] In the present invention, pharmaceutically acceptable salts include acid addition salts and base addition salts thereof. Suitable acid addition salts are formed from acids that form pharmaceutically acceptable salts. Suitable base addition salts are formed from bases that form pharmaceutically acceptable salts. For a review of suitable salts, see, for example, "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 the present invention are known to those skilled in the art. "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobromides, sulfates, nitrates, and phosphates; organic acid salts include, but are not limited to, formate, acetate, 2,2-dichloroacetate, trifluoroacetate, propionate, caproate, octanoate, decanoate, undecylenate, glycolate, gluconate, lactate, sebacate, 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-aminosalicylate, and naphthalene disulfonate. These salts can be prepared by methods known in the art. "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that retain the biological effectiveness 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, aluminum salts, and the like. Preferred inorganic salts are ammonium salts, sodium salts, calcium salts, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines, including natural 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, methylglucamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like.Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline hexyl caffeine. These salts can be prepared by methods known in the art.

[0083] In the present invention, unless otherwise indicated, "ester" refers to an ester derived from a compound described herein, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compound of the present invention in the form of a free acid or alcohol). The compound of the present invention itself may also be an ester.

[0084] The compounds of the present invention may exist in the form of solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0085] Those skilled in the art will appreciate that, since nitrogen requires an available lone pair of electrons to be oxidized to oxides, not all nitrogen-containing heterocycles are capable of forming nitrogen oxides. Those skilled in the art will recognize nitrogen-containing heterocycles that are capable of forming nitrogen oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming nitrogen oxides. Synthetic methods for preparing nitrogen oxides of heterocycles and tertiary amines are well known to those skilled in the art and include oxidizing heterocycles and tertiary amines with peroxyacids such as Peracetic Acid and Metachloroperbenzoic Acid (mCPBA), hydrogen peroxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate, and dioxirane such as dimethyldioxirane. These methods for preparing nitrogen oxides have been extensively described and reviewed in the literature, see for example: TL Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp 748-750 (AR Katritzky and AJ Boulton, Eds., Academic Press); and GWH Cheeseman and ESGWerstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp 390-392 (AR Katritzky and AJ Boulton, Eds., Academic Press).

[0086] As used herein, "metabolite" refers to a substance formed in vivo upon administration of a compound of the present invention. Metabolites of a compound can be identified using techniques known in the art, and their activity can be characterized using assays. Such products can be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, and the like of the administered compound. Therefore, the present invention includes metabolites of the compounds of the present invention, including compounds produced by contacting a compound of the present invention with a mammal for a period of time sufficient to produce a metabolic product thereof.

[0087] In the present invention, "prodrug" refers to certain derivatives of the compounds of the present invention that can be converted into compounds of the present invention having the desired activity by, for example, hydrolytic cleavage when administered to the body or thereon. Typically, such prodrugs will be functional group derivatives of the compound that are easily converted into the desired therapeutically active compound in vivo. Further information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems", Volume 14, ACS Symposium Series (T. Higuchi and V. Stella). Prodrugs of the present invention can be prepared, for example, by replacing appropriate functional groups present in the compounds of the present invention with certain moieties known to those skilled in the art as "pro-moieties" (e.g., as described in "Design of Prodrugs", H. Bundgaard (Elsevier, 1985)).

[0088] As used herein, a "pharmaceutical composition" refers to a formulation of a compound of the present invention and a medium generally accepted in the art for delivering a biologically active compound to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of the active ingredient, and thereby exert its biological activity.

[0089] In this application, "pharmaceutically acceptable carrier" includes but is not limited to any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved or accepted by relevant governmental regulatory authorities for use in humans or livestock.

[0090] As used herein, the terms "drug combination," "drug combination," "combination therapy," "administration of an additional therapy," "administration of an additional therapeutic agent," and the like refer to a drug therapy obtained by mixing or combining more than one active ingredient, and include both fixed and non-fixed combinations of the active ingredients. The term "fixed combination" refers to the simultaneous administration of at least one compound described herein and at least one synergistic agent to a patient as a single entity or single dosage form. The term "non-fixed combination" refers to the simultaneous administration of at least one compound described herein and at least one synergistic agent to a patient as separate entities, either in combination or sequentially at variable intervals. This also applies to cocktail therapies, e.g., administration of three or more active ingredients.

[0091] In the present invention, unless otherwise specified, "tumor" includes but is not limited to leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, squamous cell lung cancer, lung adenocarcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, cervical cancer, ovarian cancer, intestinal cancer, rhinitis cancer, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, oral cancer and other diseases.

[0092] As used herein, unless otherwise indicated, "treating" or "treating" means reversing, alleviating, inhibiting the progression of, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition.

[0093] On the basis of not violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0094] FIG1 is a graph showing the changes in tumor volume in the MDA-MB-436 nude mouse model after administration of compounds 1 and 6.

[0095] FIG2 is a graph showing the changes in tumor volume in the MDA-MB-436 nude mouse model after administration of compounds 1, 6, and 21.

[0096] Figure 3 shows the changes in tumor volume in gastric cancer PDX model mice after the combination of compound 6 and carboplatin.

[0097] Figure 4 shows the tumor volume of the SUM14PT nude mouse subcutaneous transplant tumor model after the combination of compound 6 and carboplatin. DETAILED DESCRIPTION

[0098] The scheme of the present invention will be explained below with reference to the examples. Those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. Where specific techniques or conditions are not specified in the examples, the techniques or conditions described in the literature in this area or the product specifications shall be followed.

[0099] The reagents and raw materials used in the examples of the present invention are all commercially available.

[0100] Table 1 Abbreviations and their meanings in the present invention

[0101] The structures of the compounds of the present invention were determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were performed using a Bruker AVANCE-400 nuclear magnetic 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 measured at 10 -6 The units are given in ppm.

[0102] MS was measured using an Agilent SQD (ESI) mass spectrometer (manufacturer: Agilent, signal: 6110).

[0103] HPLC analysis was performed using an Agilent 1200DAD high pressure liquid chromatograph (Sunfirc C18, 150×4.6 mm, 5 μm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18, 150×4.5 mm, 5 μm column).

[0104] The thin layer chromatography silica gel plate used was Qingdao Ocean GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) had a specification of 0.15mm-0.2mm, and the specification used for thin layer chromatography separation and purification products was 0.4mm-0.5mm silica gel plate.

[0105] Column chromatography generally uses Qingdao Ocean 100-200, 200-300 mesh silica gel as the carrier.

[0106] Unless otherwise specified, all reactions in the following examples were conducted under an argon or nitrogen atmosphere. Argon or nitrogen atmosphere refers to the reaction flask being connected to an approximately 1 L argon or nitrogen balloon. Hydrogen atmosphere refers to the reaction flask being connected to an approximately 1 L hydrogen balloon. The hydrogenation reaction was typically performed by evacuating the flask and then filling it with hydrogen, repeating this process three times.

[0107] Intermediate preparation

[0108] Intermediate int-1: N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0109] 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 of dioxane, 3 ml of anhydrous ethanol, and 4 ml of water. The nitrogen atmosphere was then replaced three times and the mixture was reacted at 90°C for 2 h under nitrogen. After TLC analysis, the reaction mixture was cooled to room temperature, 30 ml of dichloromethane and 20 ml of water were added, and the layers were separated in a separatory funnel. The aqueous phase was extracted twice with dichloromethane, and the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and spin-dried. The resulting crude product was purified by column chromatography to yield compound int-1c (1 g, white solid).

[0110] Step 2: Int-1c (1 g, 3 mmol), aqueous methylamine (5 g, 161.3 mmol), and anhydrous methanol (20 ml) were added to a 100 ml reaction flask and stirred overnight at room temperature. After completion of the reaction as monitored by TLC, the reaction mixture was concentrated under reduced pressure to dryness to afford int-1d (0.8 g, white solid).

[0111] Step 3: Add compound int-1d (0.5 g, 1.5 mmol) to 10 ml of anhydrous methanol, followed by 10 ml of 4 mol / L hydrochloric acid and dioxane solution. Stir at room temperature for 0.5-1 h. After the reaction is complete as monitored by TLC, the reaction solution is concentrated under reduced pressure to dryness to obtain compound int-1 (0.5 g, white solid).

[0112] Example 1: 1'-(7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0113] Step 1: Add compound 1a (20 g, 95.1 mmol) and selenium dioxide (16 g, 144 mmol) to a 250 ml reaction flask, then add 120 ml of 1,4-dioxane. Heat to 110°C and stir overnight. After the reaction is complete as monitored by TLC, filter the reaction mixture, rinse the residue with ethyl acetate, combine the filtrates, and concentrate by rotary evaporation. The crude product is purified by column chromatography to yield compound 1b (16 g, yellow solid). LC-MS: ESI [M+H] + =225.2.

[0114] Step 2: Add sodium hydride (6.86 g, 171.4 mmol) and 60 ml of 1,4-dioxane to a 250 ml reaction flask, then replace the nitrogen atmosphere three times. Cool to 0°C and slowly add triethyl 2-phosphonobutyrate (43.2 g, 171.4 mmol) dropwise under nitrogen. Stir and react at 0°C for 10 minutes. Warm to room temperature, stir and react for 10 minutes, then heat to 40°C, stir and react 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 dropwise and stir and react at -78°C for 1 minute. After TLC monitoring, quench the reaction by slowly adding ice-cold saturated ammonium chloride aqueous solution. Extract three times with 150 ml of ethyl acetate. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product was purified by column chromatography to give compound 1c (13.26 g, brown liquid). LC-MS: ESI [M+H] + =323.3.

[0115] Step 3: Compound 1c (13.26 g, 41.1 mmol) was added to 100 ml of anhydrous ethanol, followed by the addition of Pd / C (1.33 g, 10%), and the reaction was stirred at room temperature overnight. After the reaction was complete as monitored by LC-MS, the reaction solution was filtered, the residue was rinsed with a large amount of ethanol, the filtrates were combined and concentrated by rotary evaporation. A 4 mol / L hydrochloric acid solution in 1,4-dioxane (50 ml) was added, and the mixture was stirred at room temperature for 30 minutes. Diethyl ether was added to precipitate a large amount of solid, which 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).

[0116] Step 4: Compound 1d (7.32 g, 29.5 mmol) and 120 ml of 1,4-dioxane were added to a 250 ml reaction flask, followed by DDQ (7.38 g, 32.5 mmol). The mixture was refluxed overnight. After completion of the reaction as monitored by LC-MS, the reaction solution was concentrated by rotary evaporation, saturated sodium bicarbonate solution was added, and the mixture was stirred for 1 hour. The mixture was filtered, and the residue was rinsed with water and then washed with a small amount of diethyl ether. After drying, compound 1e (2.31 g, yellow solid) was obtained. LC-MS: ESI [M+H] + =247.3.

[0117] Step 5: Compound 1e (2.0 g, 8.1 mmol) and 60 ml of tetrahydrofuran were added to a 150 ml reaction flask, cooled to 0°C, and then a 2.5 mol / L solution of lithium aluminum hydride in tetrahydrofuran (6.48 ml, 16.2 mmol) was added. The mixture was allowed to react at 0°C for 2 hours. After TLC monitoring, the reaction was quenched by adding 5 ml of water, dried over a large amount of anhydrous sodium sulfate, filtered, and the residue rinsed with a large amount of dichloromethane. The filtrates were combined and concentrated by rotary evaporation. After drying, compound 1f (1.2 g, white solid) was obtained. 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).

[0118] Step 6: Compound 1f (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 temperature was cooled to 0°C, and thionyl chloride (0.87 ml, 12 mmol) was added dropwise. The mixture was allowed to react at 0°C for 1 hour. After completion of the reaction as monitored by TLC, the reaction solution was concentrated by rotary evaporation, and the crude product was purified by column chromatography to yield compound 1g (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).

[0119] Step 7: Compound int-1 (0.05 g, 0.23 mmol), 1g (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 and stirred at 80 ° C for 2 h. After the reaction was completed 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).

[0120] Example 2: 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0121] Step 1: Weigh int-1c (1.4 g, 4.5 mmol), add 20 mL of MeOH to dissolve, add 5 mL of water, add lithium hydroxide (570 mg, 13.5 mmol), and react at room temperature for 12 h. The reaction is monitored by TLC. After the reaction is complete, 2 M HCl is added to adjust the pH to 6. EA (3 x 25 mL) is added for extraction. The organic phases are combined, dried over anhydrous sodium sulfate, and dried in vacuo to obtain product 2a (460 mg, light yellow solid).

[0122] Step 2: Weigh 2a (230 mg, 0.7 mmol), EDCI (191 mg, 1 mmol), HOBT (100 mg, 1 mmol) 2 mL DMF, N-methylmorpholine (250 mg, 2.8 mmol), d3-methylamine hydrochloride (49 mg, 0.7 mmol), and react at room temperature for 12 h. Monitor by TLC. After the starting materials are completely consumed, dilute with water, extract with EA, and wash the organic phase with water 5 times, dry over anhydrous sodium sulfate, and spin dry to give the crude product 2b (150 mg, light yellow oily liquid).

[0123] 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 allowed to react at room temperature for 12 h. The reaction was monitored by TLC. After the reaction was complete, potassium carbonate was added and stirred for 30 minutes. The potassium carbonate was removed by filtration to obtain the crude product of compound 2c (270 mg, yellow-green solid).

[0124] Step 4: 1f (40.6 mg, 0.2 mmol) was weighed, and 1 mL of DCM and DMF (15 mg, 0.02 mmol) were added. Thionyl chloride (70 mg, 0.6 mmol) was added at 0°C and allowed to react for 30 minutes. The mixture was then allowed to react at room temperature for 30 minutes. TLC was used to monitor the reaction. After the starting material was completely consumed, the mixture was dried under vacuum and then 2c (44.2 mg, 0.2 mmol) and DIPEA (180 mg, 1.4 mmol) were added. KI (10 mg, 0.06 mmol) was then added, followed by 2 mL of acetonitrile. The temperature was raised to 80°C and the reaction was allowed to react for 2 h. The reaction was monitored by TLC. After completion, 2 mL of saturated sodium bicarbonate solution was added, and the mixture was extracted with EA (2 x 3 mL). The organic phases were combined and purified by preparative TLC to yield 2 (12 mg, light 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).

[0125] 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

[0126] Step 1: Compound 1f (0.4 g, 2.0 mmol) and dichloromethane (20 mL) were added to a 50 mL reaction flask. The temperature was cooled to 0°C, and Dess-Martin periodinane (1.7 g, 4.0 mmol) was added. The mixture was allowed to react at 0°C for 1.5 hours. After completion of the reaction, the mixture was filtered through celite, and the residue was rinsed with dichloromethane. The filtrate was concentrated by rotary evaporation and purified by column chromatography to obtain compound 3a (0.38 g, light 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).

[0127] Step 2: Weigh compound 3a (0.2 g, 1 mmol) into a reaction flask, add 15 mL of ether and 4 mL of methanol, cool to 0°C, add sodium borodeuteride (46 mg, 1.1 mmol), and warm to room temperature for 1 hour. After TLC, quench with a small amount of water, concentrate by rotary evaporation, slurry with a small amount of ice water, filter, and dry. Compound 3b (110 mg, white solid) was obtained; LC-MS: ESI [M+H] + =206.2.

[0128] 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 temperature was cooled to 0°C, and thionyl chloride (88 mg, 0.75 mmol) was added dropwise. The mixture was allowed to react at 0°C for 1 hour. After completion of the reaction as monitored by TLC, the reaction solution was concentrated by rotary evaporation. The crude product was purified by column chromatography to afford compound 3c (103 mg, white solid); LC-MS: ESI [M+H] + =224.7.

[0129] 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 and stirred at 80°C for 2 hours. After the reaction was completed 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, light yellow solid); LC-MS: ESI [M+H] + =405.5; 1 H 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).

[0130] 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

[0131] 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 to react overnight. After TLC detection, cool to room temperature, slowly add ice water to dilute the reaction solution, extract three times with ethyl acetate, combine the organic phases, wash with water, wash with saturated sodium bicarbonate aqueous solution, dry over anhydrous sodium sulfate, filter, and spin dry. The resulting crude product is purified by column chromatography to obtain compound 4b (65 g, light yellow solid); LC-MS: ESI [M+H] + =238.9.

[0132] 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 and extracted three times with ethyl acetate and water. The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product was purified by column chromatography to obtain compound 4d (37 g, orange-red solid); LC-MS: ESI [M+H] + =336.1.

[0133] Step 3: Compound 4d (26 g, 78 mmol) was added to 200 ml of anhydrous methanol and 5 ml of water, followed by the addition of ammonium chloride (35 g, 621 mmol). The temperature was lowered to 0°C, and zinc powder (43 g, 621 mmol) was slowly added. The temperature was raised to room temperature and the reaction was allowed to react for 1 hour. After TLC monitoring, the reaction was completed, and the mixture was filtered. The filtrate was concentrated under reduced pressure and 30 ml of a 4 mol / L hydrochloric acid solution in dioxane was added. The mixture was allowed to react at room temperature for 1 hour. After TLC monitoring, the reaction was completed, and petroleum ether was added to precipitate a solid, which was filtered and dried to obtain compound 4e (16 g, off-white solid). LC-MS: ESI [M+H] + =274.1.

[0134] 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 TLC monitoring, the reaction solution was concentrated under reduced pressure and quenched by adding saturated sodium bicarbonate aqueous solution. The mixture was extracted three times with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by column chromatography to obtain compound 4f (7.9 g, brown solid); LC-MS: ESI [M+H] + =272.1.

[0135] Step 5: 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) were weighed, 15 ml of dioxane was added, the nitrogen atmosphere was replaced, and the temperature was raised to 80°C for overnight reaction. After TLC monitoring, the reaction was complete, and water and ethyl acetate were added and extracted three times. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by column chromatography to obtain compound 4g (0.4 g, light yellow solid); LC-MS: ESI [M+H] + =223.2.

[0136] Step 6: Compound 4g (0.4g, 0.8mmol) was added to 10ml of aqueous hydrobromic acid solution and the temperature was raised to 80°C for 3 hours. After the reaction was complete as monitored by TLC, saturated aqueous sodium bicarbonate solution was added to quench the reaction. The mixture was extracted three times with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The crude product was purified by column chromatography to obtain compound 4h (0.24g, light yellow solid); LC-MS: ESI [M+H] + =286.1.

[0137] 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 and 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, light yellow solid). LC-MS: ESI [M+H] + =425.2; 1H 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).

[0138] 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

[0139] 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), and react at room temperature for 12 h. Monitor by TLC. After the starting materials are completely consumed, dilute with water, extract with EA, and wash the organic phase with water five times, dry over anhydrous sodium sulfate, and spin dry to give the crude product 5a (180 mg, yellow oily liquid).

[0140] 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 allowed to react at room temperature for 12 h and monitored by TLC. After the reaction was complete, potassium carbonate was added and stirred for 30 minutes. The potassium carbonate was then removed by filtration to obtain the crude product of compound 5b (180 mg, yellow-brown solid).

[0141] 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, transfer to room temperature and react for 30 minutes. Monitor by TLC. After the starting material is completely consumed, vacuum spin dry, then add 5b (44.2 mg, 0.2 mmol) and DIPEA (180 mg, 1.4 mmol). KI (10 mg, 0.06 mmol) and 2 mL of acetonitrile are added. After heating to 80°C, react for 2 hours. Monitor the reaction by TLC. After the reaction is complete, add 2 mL of saturated sodium bicarbonate solution and extract with EA (2 x 3 mL). The organic phases are combined and purified by preparative TLC to obtain 5 (8 mg, light 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).

[0142] Example 6: 1'-((7-(ethyl-d5)-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0143] Step 1: Weigh 60% sodium hydride (16 g, 406 mmol) and add DME (200 mL). Replace the nitrogen atmosphere three times, cool the temperature to 0°C, and add compound 6a (76 g, 338 mmol) dropwise. Warm the reaction to room temperature for 2 hours. Add deuterated bromoethane (50 g, 439 mmol) dropwise, and heat the temperature to 60°C for 3 hours. After monitoring the reaction progress by HPLC, slowly pour the reaction solution into ice water to quench the reaction. Extract with ethyl acetate three times. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin-dry. This yields crude compound 6c (83 g, colorless liquid).

[0144] Step 2: Add 60% sodium hydride (13g, 322mmol) to the reaction flask, add 200ml of tetrahydrofuran, and then replace nitrogen three times. Cool to 0°C, slowly add compound 6c (83g, 322mmol), stir and react at 0°C for 10 minutes, warm to room temperature and stir and react for 10 minutes, then warm to 40°C and stir and react for 5 minutes, and cool the reaction to -78°C. Slowly add a solution of compound 1b (48g, 215mmol) dissolved in 200ml of tetrahydrofuran, keep stirring and react at -78°C for 1 hour. After TLC monitoring, the reaction solution was slowly added to an ice-cold saturated ammonium chloride aqueous solution to quench, extracted three times with ethyl acetate, combined the organic phases, dried over anhydrous sodium sulfate, filtered, and spin-dried. The resulting crude product was purified by column chromatography to obtain compound 6d (41g, yellow-green liquid). LC-MS: ESI[M+H] + =328.4.

[0145] 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%), followed by replacement of hydrogen three times, and the reaction was stirred at room temperature overnight. After the reaction was complete as monitored by LC-MS, the reaction solution was filtered, the residue was rinsed with a large amount of ethanol, the filtrates were combined and concentrated by rotary evaporation. A 4 mol / L hydrochloric acid solution in 1,4-dioxane (12 ml) was added, and the mixture was stirred at room temperature for 30 minutes. Ether was added to precipitate a large amount of solid, which was filtered and dried to obtain compound 6e (0.51 g, white solid); LC-MS: ESI [M+H] + =254.3.

[0146] Step 4: Compound 6e (0.51 g, 2.0 mmol) and 15 mL of 1,4-dioxane were added to a 50 mL reaction flask, followed by DDQ (0.50 g, 2.2 mmol). The mixture was refluxed overnight. After completion of the reaction as monitored by LC-MS, the reaction solution was concentrated by rotary evaporation, saturated aqueous sodium bicarbonate solution was added, and the mixture was stirred for 1 hour. The mixture was filtered, and the residue was rinsed with water and then washed with a small amount of diethyl ether. After drying, compound 6f (0.3 g, yellow solid) was obtained. LC-MS: ESI [M+H] + =252.3.

[0147] Step 5: Add compound 6f (0.3 g, 1.2 mmol) and 15 ml of tetrahydrofuran to a reaction flask, cool to 0°C, then add 2.5 mol / L lithium aluminum hydride solution in tetrahydrofuran (1.44 ml, 3.6 mmol) and react at 0°C for 2 hours. After TLC monitoring, quench the reaction by adding 1 ml of water, dry with a large amount of anhydrous sodium sulfate, filter, and rinse the filter residue with a large amount of dichloromethane. The filtrates are combined and concentrated by rotary evaporation. After drying, compound 6g (0.2 g, yellow solid) is obtained. LC-MS: ESI [M+H] + =210.3.

[0148] Step 6: Compound 6g (0.2g, 0.96mmol) was added to a 25ml reaction flask, along with 10ml of dichloromethane and 0.1ml of N,N-dimethylformamide. The temperature was cooled to 0°C, and thionyl chloride (0.35g, 2.9mmol) was added dropwise. The mixture was allowed to react at 0°C for 1 hour. After completion of the reaction as monitored by TLC, the reaction solution was concentrated by rotary evaporation, and the crude product was purified by column chromatography to yield compound 6h (0.2g, gray solid). LC-MS: ESI [M+H] + =228.7.

[0149] 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 and stirred at 85 ° C for 2 hours. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure and the crude product was purified by column chromatography to give 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).

[0150] Example 7: 1'-((7-(ethyl-d5)-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N,2-dimethyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0151] 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 of dioxane, 3 ml of anhydrous ethanol, and 4 ml of water. The nitrogen atmosphere was then replaced three times and the mixture was reacted at 90°C for 2 h. After TLC analysis of the reaction, the reaction mixture was cooled to room temperature, 30 ml of dichloromethane and 20 ml of water were added, and the layers were separated in a separatory funnel. The aqueous phase was extracted twice with dichloromethane, and the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and spin-dried. The resulting crude product was purified by column chromatography to yield compound 7b (0.7 g, white solid).

[0152] Step 2: Compound 7b (0.7 g, 26 mmol) was added to 10 ml of anhydrous methanol, followed by 20 ml of aqueous methylamine solution. The mixture was stirred at room temperature overnight. After completion of the reaction as monitored by TLC, the reaction solution was concentrated under reduced pressure to dryness to obtain compound 7c (0.5 g, white solid).

[0153] 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 hydrochloric acid and dioxane solution. The mixture was stirred at room temperature for 0.5-1 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure to dryness to obtain compound 7d (0.3 g, white solid).

[0154] Step 4: Compound 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 and 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. 1 H 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).

[0155] 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

[0156] 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 Reference 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).

[0157] Example 9: 1'-((7-(ethyl-d5)-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-2-fluoro-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0158] 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 nitrogen atmosphere was then replaced three times and the mixture was stirred at room temperature under nitrogen for 48 h. After completion of the reaction by TLC, the reaction mixture was concentrated to dryness under reduced pressure. The crude product was purified by column chromatography to yield compound 9a (2.2 g, white solid).

[0159] 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 of dioxane, 3 ml of anhydrous ethanol, and 4 ml of water. The nitrogen atmosphere was then replaced three times and the mixture was reacted at 90°C under nitrogen for 2 h. After TLC analysis of the reaction, the reaction mixture was cooled to room temperature, 30 ml of dichloromethane and 20 ml of water were added, and the layers were separated in a separatory funnel. The aqueous phase was extracted twice with dichloromethane, and the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and spin-dried. The resulting crude product was purified by column chromatography to yield compound 9b (1.2 g, white solid).

[0160] Step 3: Add 9b (1 g, 2.9 mmol), aqueous methylamine (5 g, 161.3 mmol), and anhydrous methanol (20 ml) to a 100 ml reaction flask and stir overnight at room temperature. After completion of the reaction as monitored by TLC, the reaction solution was concentrated under reduced pressure to dryness to yield compound 9c (0.8 g, white solid).

[0161] 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 hydrochloric acid and dioxane solution. The mixture was stirred at room temperature for 0.5-1 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure to dryness to obtain compound 9d (0.3 g, white solid).

[0162] Step 5: Compound 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 and 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. 1 H 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).

[0163] 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

[0164] 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 Reference Example 2. LC-MS: ESI [M+H] + =425.2. 1H NMR (400MHz, Chloroform-d) δ11.15 (s, 1H), 8.47 (d, J = 1.8 Hz, 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).

[0165] Example 11: 1'-((7-(ethyl-d5)-6-oxo-56-dihydro-1,5-naphthyridin-3-yl)methyl)-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0166] Preparation of 1'-((7-(ethyl-d5)-6-oxo-5'-dihydro-1,5-naphthyridin-3-yl)methyl)-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide Reference Example 1. LC-MS: ESI [M+H] + =412.3. 1 H NMR (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).

[0167] 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

[0168] Preparation of 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 Reference 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).

[0169] 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

[0170] Step 1: 13a (10.0 g, 39.5 mmol) was weighed into a reaction flask and dissolved in 100 mL of DMF. t-BuOK (6.6 g, 59.2 mmol) was added portionwise to the system at -10°C under nitrogen protection. After the addition, the mixture was stirred at -10°C for 1 h. Deuterated bromoethane (5.0 g, 43.4 mmol) was then slowly added dropwise. After the addition, the mixture was returned to room temperature and stirred for 3 h. 300 mL of water was added to the system and extracted with EA (500 mL × 3). The organic phases were combined and washed with saturated brine. The organic phase was separated and dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to obtain 13b (11.0 g, 97%) as a yellow oily liquid. LC-MS: ESI [M+H] + =287.2.

[0171] Step 2: 13b (11.0 g, 38.4 mmol) was weighed and added to 120 mL of a 3 / 1 EA / H2O mixed solvent. 4 M hydrochloric acid ethyl acetate solution (28.8 mL, 115.2 mmol) was added and stirred overnight. The solvent was removed by vacuum concentration and the mixture was slurried with EA to obtain 13c hydrochloride as a white solid (2.5 g, 41%). LC-MS: ESI [M+H] + =123.1.

[0172] Step 3: The hydrochloride of 13c (2.5 g, 15.8 mmol) was weighed into a reaction flask and dissolved in 30 mL of DMF. DIEA (8.1 g, 63.0 mmol) was added and 4b (4.1 g, 17.3 mmol) was added while stirring at room temperature. The mixture was stirred at room temperature for 5 h under nitrogen protection. After the reaction, 90 mL of pure water was added and the mixture was extracted with EA (150 mL × 2). The organic phases were combined and washed with saturated brine. The organic phase was separated and concentrated in vacuo. The organic phase was filtered through a column with PE and DCM (0-20%) to obtain 13d (4.8 g, 90%) as a yellow oily liquid. LC-MS: ESI [M+H] + =340.0.

[0173] Step 4: 13d (4.5 g, 13.2 mmol) was dissolved in 80 mL of glacial acetic acid. Reduced Fe powder (4.4 g, 79.4 mmol) was slowly added to the system with stirring at room temperature. After the addition, the mixture was stirred at 70°C for 1 h. The mixture was filtered while hot and the filter cake was washed with a mixed solvent of DCM and MeOH. The filtrate was concentrated in vacuo and purified by column chromatography with PE and EA (0-25%) to give 13e (2.1 g, 57%) as a white solid. LC-MS: ESI [M+H] + =278.0.

[0174] Step 5: 13e (2.1 g, 7.6 mmol) was weighed into a reaction flask, 30 mL of DCM was added and stirred, and then DDQ (2.1 g, 9.1 mmol) was slowly added. After the addition, the mixture was stirred at room temperature overnight. After the reaction, saturated aqueous NaHCO₃ was added to the system. The layers were separated, and the aqueous phase was extracted with a mixture of DCM and MeOH (5:1) (30 mL x 3). The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, concentrated, and purified by column chromatography with DCM and MeOH (0-3%) to obtain 13f (2.0 g, 96%) as a white solid. LC-MS: ESI [M+H] + =276.0.

[0175] Step 6: 13f (1.2 g, 5.3 mmol), (tributyltin)methanol (2.8 g, 8.7 mmol) and XPhos-Pd-G2 (416.4 mg, 0.4 mmol) were weighed into a reaction flask, and 30 mL of dioxane was added to replace the nitrogen atmosphere 3-5 times. Then, the mixture was stirred at 90°C under nitrogen for 4 h. After the reaction, the solvent was concentrated in vacuo and purified by column chromatography with DCM and MeOH (0-5%) to obtain 13 g (946.0 mg, 76%) of a white solid. LC-MS: ESI [M+H] + =228.1; 1H 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).

[0176] Step 7: Weigh 13 g (650 mg, 2.9 mmol) into a reaction flask and dissolve in 20 mL of DCM. Slowly add Dess-Martin oxidant (1.5 g, 3.4 mmol) at 0°C. Maintain stirring at 0°C for 0.5 h. After the reaction is complete, add saturated aqueous NaHCO₃ solution to the system. Stir for 10 min, separate the layers, and concentrate the organic solvent in vacuo to dryness. Purify the mixture by column chromatography with DCM and MeOH (0-5%) to obtain 13h (610 mg, 95%) as a white solid. 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).

[0177] Step 8: Weigh int-1a (99.2 mg, 0.34 mmol) into a reaction flask. First, add 5 mL of DCM, then TEA to dissociate it. Then, add glacial acetic acid dropwise to adjust the pH to ≈ 5. Finally, add 13h (70.0 mg, 0.31 mmol) and stir at room temperature for 2 h. Sodium triacetoxyborohydride (263.5 mg, 1.24 mmol) was added to the system, and stirring continued for 2 h. Saturated aqueous NH4Cl solution was added and stirred. The phases were separated, and the aqueous phase was extracted with a mixture of DCM and MeOH (5:1) (30 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, concentrated, and prepared by HPLC and freeze-dried to obtain 13 (16 mg, 12%) as a white solid. 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).

[0178] 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

[0179] 9d (150.5 mg, 0.49 mmol) was weighed into a reaction flask and first added with 5 mL of DCM, followed by TEA to dissociate it. Glacial acetic acid was then added dropwise to adjust the pH to ≈ 5. Finally, 13h (100.0 mg, 0.44 mmol) was added and stirred at room temperature for 2 h. Sodium triacetoxyborohydride (376.4 mg, 1.78 mmol) was added to the system, and stirring was continued for 2 h. Saturated aqueous NH4Cl was added and stirred. The phases were separated, and the aqueous phase was extracted with a mixture of DCM and MeOH (5:1) (30 mL x 3). The organic phases were combined and dried over anhydrous Na2SO4. The filtrate was then filtered and concentrated. The residue was purified by HPLC and freeze-dried to yield 14 (60 mg, 30%) as a white solid. 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).

[0180] 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

[0181] Step 1: Compound 1e (2.0 g, 8.1 mmol) and 60 ml of tetrahydrofuran were added to a 150 ml reaction flask, 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 mixture was allowed to react at 0°C for 2 hours. After TLC monitoring, the reaction was quenched by adding 5 ml of water, dried over a large amount of anhydrous sodium sulfate, filtered, and the residue rinsed 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.

[0182] 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 temperature was cooled to 0°C, and thionyl chloride (0.87 ml, 12 mmol) was added dropwise. The mixture was allowed to react at 0°C for 1 hour. After completion of the reaction as monitored by TLC, the reaction solution was concentrated by rotary evaporation, and the crude product was purified by column chromatography to yield compound 15b (0.52 g, gray solid). LC-MS: ESI [M+H] + =225.7.

[0183] 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 and stirred at 80°C for 2 h. After the reaction was completed 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.

[0184] Example 16: 1'-((7-(ethyl-2,2,2-d3)-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide

[0185] 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 and dissolve, and then add dropwise to the reaction system. Control the temperature at 0-5 ° C. After the addition, naturally return to the temperature and stir for 1 h. After TLC detection, the reaction system is complete. After that, 2.0 L of water is added to the reaction system, and the mixture is concentrated under reduced pressure at 40-50 ° C until no liquid is discharged. The aqueous layer is extracted twice with 4.0 L and 1.0 L of ethyl acetate respectively. The organic phases are combined and concentrated under reduced pressure at 40-50 ° C to constant weight to obtain compound 16b (480 g, white solid).

[0186] Step 2: Weigh 16b (475 g, 1.93 mol), add 2.85 L of DMF, add 2.85 L of DMF-DMA, raise the temperature to 100°C after the addition, stir for 2 h, and after TLC detection, concentrate the reaction solution at 70-80°C until no liquid is discharged. Then add 10.0 L of water, stir at 20-30°C for 2 h, filter, and dry the filter cake at 60-70°C under reduced pressure to constant weight to obtain compound 16c (612 g, red solid).

[0187] Step 3: Weigh 16c (500 g, 1.91 mol), add 2.56 L of tetrahydrofuran, weigh sodium periodate (805.0 g, 3.72 mol) and add it to 2.56 L of water to dissolve it. Control the temperature at 25-30°C and add the sodium periodate aqueous solution dropwise to the reaction system. After the addition, control the temperature at 25-30°C and stir for 2-4 h. After TLC detection, add 2.56 L of water and 4.0 L of ethyl acetate. Stir and separate the layers. The aqueous layer is washed twice with 2.0 L of ethyl acetate. The combined organic layers are washed with 4.0 L of saturated sodium thiosulfate solution and 4.0 L of saturated brine in sequence. After separation, the organic layer is concentrated under reduced pressure at 40-50°C to constant weight to obtain compound 16d (500 g, oil).

[0188] Step 4: Weigh 16d (500.0 g, 1.69 mol) and 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 batches. After the addition, heat to reflux and react for 1-2 h. After TLC detection, the reaction solution is concentrated under reduced pressure at 40-50 ° C to constant weight, 15.0 L of ethyl acetate is added, and the pH is adjusted to 7-8 with saturated sodium bicarbonate solution. Filter, and concentrate the organic layer under reduced pressure at 40-50 ° C to constant weight. Purify by column chromatography to obtain compound 16e (230.0 g, white solid).

[0189] Step 5: Weigh 16e (80.0 g, 257.1 mmol), add 960 ml of tetrahydrofuran, cool to 0°C in an ice-water bath, add diisopropylaluminum hydride (1 M, 771.3 ml) dropwise, and return to 20-25°C for reaction. After the reaction is complete, LC-MS detection indicates that the reaction is complete. Add 800 ml of saturated ammonium chloride solution dropwise to the reaction system, filter, and spin-dry the filtrate. Add 80 ml of dichloromethane to pulp, 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).

[0190] Step 6: 16f (45.0 g, 167.2 mmol), DIPEA (64.8 g, 501.6 mmol), and sodium iodide (5.0 g, 33.4 mmol) were weighed and added to 450 ml of tetrahydrofuran. MOMCl (26.9 g, 334.4 mmol) was added dropwise with stirring. After the addition, the temperature was raised to 45-50°C and stirred for 3 h. After the reaction was complete by TLC, the temperature was lowered to 20-25°C, 500 ml of water was added, the layers were separated, the aqueous layer was extracted with 300 ml of ethyl acetate, and the combined organic layers were concentrated to dryness under reduced pressure at 40-50°C. Compound 16g (34.8 g, yellow solid) was obtained by column chromatography.

[0191] Step 7: Weigh 16 g (39.8 g, 127.1 mmol), add triethylamine (15.5 g, 152.5 mmol), Pd(dppf)Cl2 (9.22 g, 12.7 mmol), add 800 ml of methanol, replace with nitrogen three times, replace with carbon monoxide three times, raise the temperature to 60 ° C and react for 12 h. After LC-MS detection, the reaction solution is concentrated under reduced pressure at 40-50 ° C to remove the solvent, and purified by column chromatography to obtain compound 16h (32.0 g, white solid).

[0192] Step 8: Weigh 16h (32.0 g, 109.5 mmol), add 384 ml of tetrahydrofuran, cool to 0°C in an ice-water bath, add diisopropylaluminum hydride (1 M, 328.5 mmol) dropwise, and return to 20-25°C for reaction. After the reaction is complete, LC-MS detection is performed. To the reaction system, 320 ml of saturated ammonium chloride solution and 320 ml of ethyl acetate are added dropwise, and the mixture is filtered. The filtrate is concentrated under reduced pressure at 40-50°C to obtain compound 16i (28.6 g, yellow solid).

[0193] 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 Dess-Martin periodinane (55.0 g, 129.8 mmol) in batches, return to 20-25°C after addition, and react for 2 h. After TLC detection, add 300 ml of a saturated sodium bicarbonate: saturated sodium thiosulfate = 1:1 solution, wash twice, extract the aqueous layer with 300 ml of dichloromethane, combine the organic layers, concentrate under reduced pressure at 40-50°C, and purify by column chromatography to obtain compound 16j (23.4 g, yellow solid).

[0194] Step 10: Weigh 16j (23.4 g, 89.1 mmol), add 234 ml of tetrahydrofuran, cool to -40°C in a dry ice bath, add deuterated methylmagnesium iodide (1 M, 106.9 mmol) dropwise, and return to the temperature naturally after addition. Stir for 1 h. After TLC detection, the reaction is complete, and then add 234 ml of saturated ammonium chloride solution and 234 ml of water to quench the reaction. Stir and separate the layers. The organic layers are combined and concentrated under reduced pressure at 40-50°C to obtain compound 16k (25.6 g, oil).

[0195] Step 11: Weigh 16k (24.7 g, 88.1 mmol), add 247 ml of dichloromethane, stir to dissolve, cool to 0°C in an ice-water bath, add Dess-Martin periodinane (44.8 g, 105.7 mmol) in batches, return to 20-25°C after addition, and react for 2 h. After TLC detection, add 300 ml of a saturated sodium bicarbonate:saturated sodium thiosulfate = 1:1 solution, wash twice, extract the aqueous layer with 300 ml of dichloromethane, combine the organic layers, concentrate under reduced pressure at 40-50°C to dryness, and purify by column chromatography to obtain compound 16l (23.2 g, yellow solid).

[0196] Step 12: Weigh 16l (5.0 g, 17.9 mmol) and p-toluenesulfonyl hydrazide (3.3 g, 17.9 mmol), add 100 ml of methanol, and react at 20-25°C for 16 h. After the reaction is complete as detected by TLC, the reaction solution is filtered and the filter cake is dried under reduced pressure at 40-50°C to constant weight to obtain compound 16m (7.20 g, white solid).

[0197] Step 13: Weigh 16m (4.50 g, 10.0 mmol), add 45 ml of dichloromethane, cool to 0°C in an ice-water bath, add diisopropylaluminum hydride (1 M, 20 ml) dropwise, and return to 20-25°C after the addition to react. After the reaction is complete as detected by LC-MS, add 20 ml of saturated ammonium chloride solution dropwise to the reaction system, filter, and the filtrate is spin-dried and purified by column chromatography to obtain compound 16n (540 mg, oil).

[0198] Step 14: 16n (400.0 mg, 1.51 mmol) was weighed, 4 ml of dioxane was added, 4 ml of 48% HBr / H2O was added, and the temperature was raised to 80°C for 2 h. After the reaction was complete, the pH was adjusted to 7-8 with saturated sodium carbonate solution, filtered, and purified by filter cake column chromatography to obtain compound 16o (200 mg, white solid).

[0199] Step 15: Weigh 16o (100.0 mg, 0.48 mmol), add 3 ml of toluene, add DMF (3.5 mg, 0.048 mmol), cool to 0°C in an ice-water bath, add thionyl chloride (69.0 g, 0.48 mmol) dropwise, and after the addition, return the temperature to 20-25°C for reaction. After TLC detection, the reaction is complete, and the solvent is removed by concentration under reduced pressure to obtain compound 16p (108 mg, yellow solid).

[0200] Step 16: 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) were added to 3 ml of acetonitrile, and the temperature was raised to 80°C for 2 h. After the reaction was complete as determined by LC-MS, the temperature was lowered to room temperature and the mixture was dried by spin drying. The crude product was 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).

[0201] 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-carbonitrile

[0202] 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 of dioxane, 3 ml of anhydrous ethanol, and 4 ml of water. The nitrogen atmosphere was then replaced three times and the mixture was reacted at 90°C for 2 h. After TLC analysis of the reaction, the reaction mixture was cooled to room temperature, 30 ml of dichloromethane and 20 ml of water were added, and the layers were separated in a separatory funnel. The aqueous phase was extracted twice with dichloromethane, and the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and spin-dried. The resulting crude product was purified by column chromatography to yield compound 17b (0.7 g, white solid).

[0203] 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 hydrochloric acid-dioxane solution. The mixture was stirred at room temperature for 0.5-1 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure to dryness to obtain compound 17c (0.2 g, white solid).

[0204] 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 and 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).

[0205] Example 18: 3-(ethyl-2,2,2-d3)-7-((5-fluoro-3',6'-dihydro-[2,4'-bipyridinyl]-1'(2'H)-yl)methyl)-1,5-naphthyridin-2(1H)-one

[0206] Preparation of 3-(ethyl-2,2,2-d3)-7-((5-fluoro-3',6'-dihydro-[2,4'-bipyridyl]-1'(2'H)-yl)methyl)-1,5-naphthyridin-2(1H)-one Reference Example 17. LC-MS: ESI [M+H] + =368.2. 1 H 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).

[0207] Example 19: 3-(Ethyl-d5)-8-fluoro-7-((5-fluoro-3',6'-dihydro-[2,4'-bipyridyl]-1'(2'H)-yl)methyl)quinoxalin-2(1H)-one

[0208] 18c (122.6 mg, 0.49 mmol) was weighed into a reaction flask and first added with 5 mL of DCM, followed by TEA to dissociate it. Glacial acetic acid was then added dropwise to adjust the pH to ≈ 5. Finally, 13h (100.0 mg, 0.44 mmol) was added and stirred at room temperature for 2 h. Sodium triacetoxyborohydride (376.4 mg, 1.78 mmol) was added to the system, and stirring was continued for 2 h. Saturated aqueous NH4Cl solution was then added and stirred. The mixture was separated, and the aqueous phase was extracted with a mixture of DCM and MeOH (5:1) (30 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, concentrated, and lyophilized to obtain 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).

[0209] 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-carbonitrile

[0210] 17c (126.1 mg, 0.49 mmol) was weighed into a reaction flask and first added with 5 mL of DCM, followed by TEA to dissociate it. Glacial acetic acid was then added dropwise to adjust the pH to ≈ 5. Finally, 13h (100.0 mg, 0.44 mmol) was added and stirred at room temperature for 2 h. Sodium triacetoxyborohydride (376.4 mg, 1.78 mmol) was added to the system, and stirring was continued for 2 h. Saturated aqueous NH4Cl was added and stirred. The phases were separated, and the aqueous phase was extracted with a mixture of DCM and MeOH (5:1) (30 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, concentrated, and prepared by HPLC and freeze-dried to obtain 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).

[0211] 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

[0212] Step 1: Compound 21a (6.0 g, 49.2 mmol) was added to 50 ml of tetrahydrofuran, cooled to -78°C, and LDA (2M, 29.5 ml, 59.0 mmol) was added dropwise at a temperature of -60 to -78°C. After the addition, the mixture was stirred for 15 min. 40 ml of a tetrahydrofuran solution of 21b (19.3 g, 54.1 mmol) was added dropwise. After the addition, the mixture was naturally warmed to room temperature, stirred for 2 h, and sampled. After TLC detection, the reaction was complete, and 60 ml of a saturated aqueous ammonium chloride solution was added and stirred. The layers were separated, and 100 ml of ethyl acetate was added to the aqueous layer for extraction. The layers were separated, and the combined organic layers were washed with 100 ml of saturated brine. The layers were separated, and the organic phase was concentrated under reduced pressure to remove the solvent to obtain compound 21c (20.0 g, oil).

[0213] Step 2: To 100 ml of a 1,4-dioxane / water solution (10:1) were added 21c (19.3 g, 28.7 mmol), potassium carbonate (7.93 g, 57.4 mmol), and 21d (8.32 g, 31.6 mmol) (100 ml). The mixture was replaced with nitrogen three times, heated to 80°C and stirred for 2 h. After the reaction was complete as detected by LC-MS, the mixture was cooled to room temperature, 100 ml of water and 100 ml of ethyl acetate were added, the layers were separated, and the aqueous layer was extracted three times with 100 ml of ethyl acetate. The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product was purified by column chromatography to give compound 21e (4.2 g, yellow solid).

[0214] Step 3: 21e (4.2 g, 13.0 mmol) and 40% methylamine aqueous solution (5.1 g, 65.2 mmol) were added to 21 ml of anhydrous methanol, and the mixture was stirred at room temperature for 1 h. After the reaction was completed by LC-MS, the reaction solution was concentrated to dryness, 40 ml of 1,4-dioxane was added, and 4M / dioxane hydrochloride solution (16.3 ml, 65.2 mmol) was added dropwise. The mixture was stirred at room temperature for 2 h. After the reaction was completed by LC-MS, the mixture was filtered and dried to obtain compound 21f (3.4 g, off-white solid).

[0215] Step 4: Compound 21f (3.4 g, 11.5 mmol), DIEA (5.6 g, 46.0 mmol), compound 1g (2.4 g, 10.5 mmol), and KI (340 mg, 2.1 mmol) were added to 48 ml of acetonitrile, and the mixture was heated to 80°C and stirred for 2 h. After the reaction was complete as determined by LC-MS, 200 ml of saturated sodium bicarbonate solution was added, the mixture was stirred and filtered, and the mixture was purified by spin 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).

[0216] 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

[0217] Step 1: Compound 21c (20 g, 60 mmol), pinacol diboron (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 and reacted at 80°C overnight. After completion of the reaction, the mixture was filtered, washed with EA, and the filtrate was concentrated. The residue was purified by column chromatography to obtain compound 22a (15 g, oil).

[0218] Step 2: To 100 ml of a 1,4-dioxane / water solution (10:1) were added 22a (15 g, 22.3 mmol), potassium carbonate (7.93 g, 57.4 mmol), and 9a (8.32 g, 31.6 mmol) (100 ml). The mixture was replaced with nitrogen three times, heated to 80°C and stirred for 2 h. After the reaction was complete as detected by LC-MS, the mixture was cooled to room temperature, 100 ml of water and 100 ml of ethyl acetate were added, the layers were separated, and the aqueous layer was extracted three times with 100 ml of ethyl acetate. The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product was purified by column chromatography to obtain compound 22b (12 g, yellow solid).

[0219] Step 3: 22b (4.2 g, 13.0 mmol) and 40% methylamine aqueous solution (5.1 g, 65.2 mmol) were added to 21 ml of anhydrous methanol, and the mixture was stirred at room temperature for 1 h. After the reaction was completed by LC-MS, the reaction solution was concentrated to dryness, 40 ml of 1,4-dioxane was added, and 4M / dioxane hydrochloride solution (16.3 ml, 65.2 mmol) was added dropwise. The mixture was stirred at room temperature for 2 h. After the reaction was completed by LC-MS, it was filtered and the filtrate was dried to obtain compound 22c (3.4 g, off-white solid).

[0220] Step 4: Compound 22c (3.4 g, 11.5 mmol), DIEA (5.6 g, 46.0 mmol), compound 1g (2.4 g, 10.5 mmol), and KI (340 mg, 2.1 mmol) were added to 48 ml of acetonitrile, and the mixture was heated to 80°C and stirred for 2 h. After the reaction was complete as determined by LC-MS, 200 ml of saturated sodium bicarbonate solution was added, the mixture was stirred and filtered, and the mixture was purified by spin 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).

[0221] 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

[0222] Step 1: To 100 ml of a 1,4-dioxane / water solution (10:1) were added 22a (13 g, 21.2 mmol), potassium carbonate (7.8 g, 55 mmol), and 7a (8.1 g, 28.6 mmol) (100 ml). The mixture was replaced with nitrogen three times, heated to 80° C. and stirred for 2 h. After the reaction was complete as detected by LC-MS, the temperature was lowered to room temperature, 100 ml of water and 100 ml of ethyl acetate were added, the layers were separated, and the aqueous layer was extracted three times with 100 ml of ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and spin-dried. The resulting crude product was purified by column chromatography to obtain compound 23a (10.6 g, yellow solid).

[0223] Step 2: 23a (3.8 g, 11.3 mmol) and 40% methylamine aqueous solution (4.5 g, 60.5 mmol) were added to 21 ml of anhydrous methanol, and the mixture was stirred at room temperature for 1 h. After the reaction was completed by LC-MS, the reaction solution was concentrated to dryness, 40 ml of 1,4-dioxane was added, and 4M / dioxane hydrochloride solution (15 ml, 64 mmol) was added dropwise. The mixture was stirred at room temperature for 2 h. After the reaction was completed by LC-MS, the mixture was filtered, and the filtrate was dried to give compound 23b (3.2 g, off-white solid).

[0224] Step 3: Compound 23b (3.2 g, 11.2 mmol), DIEA (5.4 g, 45.2 mmol), compound 1g (2.1 g, 10.5 mmol), and KI (320 mg, 1.9 mmol) were added to 48 ml of acetonitrile, and the mixture was heated to 80°C and stirred for 2 h. After the reaction was complete as determined by LC-MS, 200 ml of saturated sodium bicarbonate solution was added, the mixture was stirred and filtered, and the mixture was purified by spin 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).

[0225] Biological activity test:

[0226] 1. PARP-1 / 2 / 5a enzyme assay

[0227] 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), histone (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).

[0228] 1.1 PARP1 enzyme assay

[0229] 1.1.1 Buffer preparation: PBST: 1X PBS, 0.05% Tween-20, blocking solution: 1X PBS, 0.05% Tween-20, 5% BSA, reaction buffer: 50 mM Tris-HCl (pH 7.5), 0.005% Tween-20, 0.01% BSA.

[0230] 1.1.2 Coating: Prepare 50 ng / mL Histone coating solution in 1xPBS, transfer 25 μL of the coating solution to a 384-well reaction plate, and coat overnight at 4°C.

[0231] 1.1.3 Washing: After coating, discard the coating solution and wash with PBST solution. Transfer 50uL PBST to a 384-well reaction plate, let it stand for 5 minutes, discard the wash solution, refill the plate, and repeat the washing process three times. Finally, pat the reaction plate dry and wait for the next step of blocking.

[0232] 1.1.4 Blocking: Transfer 50 μL of blocking solution to a 384-well reaction plate and let it stand for 1 hour.

[0233] Washing: After blocking, discard the blocking solution and wash the plate three times with PBST solution according to step 2. Finally, pat the reaction plate dry.

[0234] 1.1.5 Prepare a 1000-fold dilution of the compound, transfer 1 μL of the compound to 199 μL of reaction buffer in a 96-well plate, mix thoroughly, and transfer 5 μL of the mixed compound to a 384-well reaction plate.

[0235] 1.1.6 Prepare a 25 / 10x 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.

[0236] 1.1.7 Prepare a 25 / 10x NAD+ solution in reaction buffer. Transfer 10 μL of NAD+ solution to a 384-well reaction plate. The final NAD+ concentration is 3.5 μM. Incubate at room temperature for 60 minutes.

[0237] 1.1.8 Prepare a 25 / 10x NAD+ solution in reaction buffer. Transfer 10 μL of NAD+ solution to a 384-well reaction plate. The final NAD+ concentration is 3.5 μM. Incubate at room temperature for 60 minutes.

[0238] 1.1.9 Washing: After the reaction is completed, discard the reaction solution and wash the plate three times with PBST solution according to the method in step 2. Finally, pat the reaction plate dry.

[0239] 1.1.10 Dilute the primary antibody (anti-Poly / Mono-ADP Ribose Rabbit mAb) 2000-fold with blocking buffer, add 20 μL of primary antibody, and incubate at room temperature for 1.5 hours.

[0240] 1.1.11 Washing: Discard the primary antibody and wash the plate three times with PBST solution according to the method in step 2. Finally, pat the reaction plate dry.

[0241] 1.1.12 Dilute the secondary antibody (anti-rabbit IgG, HRP-linked Antibody) 2000-fold with blocking solution, add 20 μL of secondary antibody, and incubate at room temperature for 1 hour.

[0242] 1.1.13 Washing: Discard the secondary antibody and wash the plate three times with PBST solution according to step 2. Finally, pat the reaction plate dry.

[0243] 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-well reaction plate.

[0244] 1.1.15 Reading: Use Envision to read the chemiluminescence value RLU.

[0245] PARP2 enzyme assay

[0246] 1.2.1 Preparation of buffer: PBST: 1X PBS, 0.05% Tween-20, blocking solution: 1X PBS, 0.05% Tween-20, 5% BSA, reaction buffer: 50 mM HEPES (pH 7.5), 0.002% Tween-20, 0.1% BSA, 100 mM NaCl, 2 mM DTT.

[0247] 1.2.2 Coating: Prepare 100 ng / mL Histone coating solution in 1xPBS, transfer 25 μL of the coating solution to a 384-well reaction plate, and coat overnight at 4°C.

[0248] 1.2.3 Washing: After coating, discard the coating solution and wash with PBST solution. Transfer 50uL PBST to a 384-well reaction plate, let it stand for 5 minutes, discard the wash solution, refill the plate, repeat the washing three times, and finally pat the reaction plate dry and wait for the next step of blocking.

[0249] 1.2.4 Blocking: Transfer 50 μL of blocking solution to a 384-well reaction plate and let it stand for 1 hour.

[0250] 1.2.5 Washing: After blocking, discard the blocking solution and wash the plate three times with PBST solution according to step 2. Finally, pat the reaction plate dry.

[0251] 1.2.6 Prepare 25 / 10x PARP2 solution and 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 concentration of PARP2 is 1.5 nM.

[0252] 1.2.7 Prepare a 2000-fold dilution of the compound, transfer 50 nL of the compound using an echo tube, add 19.95 uL of reaction buffer to the compound, mix thoroughly, and transfer 5 uL of the mixed compound to a 384-well reaction plate.

[0253] 1.2.8 Prepare a 25 / 10x Biotin-NAD+ solution. Transfer 10 μL of Biotin-NAD+ solution to a 384-well reaction plate. The final concentration of Biotin-NAD+ is 2 μM. Incubate at room temperature for 60 minutes.

[0254] 1.2.9 Washing: After the reaction is completed, discard the reaction solution and wash the plate three times with PBST solution according to the method in step 2. Finally, pat the reaction plate dry.

[0255] 1.2.10 Prepare Stre-HRP solution by diluting with blocking buffer. Transfer 25 μL of Stre-HRP solution to the reaction plate and incubate at room temperature for 1 hour. The final concentration of Stre-HRP is 0.1 μg / mL.

[0256] 1.2.11 Washing: Discard the Stre-HRP solution and wash the plate three times with PBST solution according to step 2. Finally, pat the reaction plate dry.

[0257] 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 to a 384-well reaction plate, incubate at room temperature for 10 minutes, and add 2.5 μL of QuantaRed Stop Solution.

[0258] 1.2.13 Reading: Read the fluorescence value using Paradigm (Ex550 / Em620).

[0259] PARP5A enzyme assay

[0260] 1.3.1 Buffer preparation: PBST: 1X PBS, 0.05% Tween-20, blocking buffer: 1X PBS, 0.05% Tween-20, 5% BSA, reaction buffer: 50 mM HEPES (pH 7.5), 0.002% Tween-20, 0.1% BSA, 100 mM NaCl, 2 mM DTT.

[0261] 1.3.2 Coating: Prepare 100 ng / mL Histone coating solution in 1xPBS, transfer 25 μL of the coating solution to a 384-well reaction plate, and coat overnight at 4°C.

[0262] 1.3.3 Washing: After coating, discard the coating solution and wash with PBST solution. Transfer 50uL PBST to a 384-well reaction plate, let it stand for 5 minutes, discard the wash solution, refill the plate, repeat the washing process 3 times, and finally pat the reaction plate dry and wait for the next step of blocking.

[0263] 1.3.4 Blocking: Transfer 50 μL of blocking solution to a 384-well reaction plate and let it stand for 1 hour.

[0264] 1.3.5 Washing: After blocking, discard the blocking solution and wash the plate three times with PBST solution according to step 2. Finally, pat the reaction plate dry.

[0265] 1.3.6 Prepare a 25 / 10x PARP5A solution and transfer 10 μL of PARP5A solution to a 384-well reaction plate. For the negative control wells, transfer 10 μL of reaction buffer. The final concentration of PARP5A is 10 nM.

[0266] 1.3.7 Prepare a 2000-fold dilution of the compound, transfer 50 nL of the compound using an echo aspirator, add 19.95 uL of reaction buffer to the compound, mix thoroughly, and transfer 5 uL of the mixed compound to a 384-well reaction plate.

[0267] 1.3.8 Prepare a 25 / 10x Biotin-NAD+ solution. Transfer 10 μL of Biotin-NAD+ solution to a 384-well reaction plate. The final concentration of Biotin-NAD+ is 2 μM. Incubate at room temperature for 60 minutes.

[0268] 1.3.9 Washing: After the reaction is completed, discard the reaction solution and wash the plate three times with PBST solution according to the method in step 2. Finally, pat the reaction plate dry.

[0269] 1.3.10 Prepare Stre-HRP solution by diluting with blocking buffer. Transfer 25 μL of Stre-HRP solution to the reaction plate and incubate at room temperature for 1 hour. The final concentration of Stre-HRP is 0.1 μg / mL.

[0270] 1.3.11 Washing: Discard the Stre-HRP solution and wash the plate three times with PBST solution according to step 2. Finally, pat the reaction plate dry.

[0271] 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 to a 384-well reaction plate, incubate at room temperature for 10 minutes, and add 2.5 μL of QuantaRed Stop Solution.

[0272] 1.3.13 Reading: Read the fluorescence value using Paradigm (Ex 550 / Em 620).

[0273] The test results are shown in Table 2 below:

[0274] Table 2 PARP-1 / 2 / 5a enzyme test results

[0275] Conclusion: The compounds of the present invention have a significant inhibitory effect on PARP1 and a weaker inhibitory effect on PARP2 / 5a, indicating that the compounds of the present invention selectively inhibit PARP-1.

[0276] 2. Cell anti-proliferation activity test:

[0277] BRCA-mutant MDA-MB-436 cells were cultured in DMEM medium supplemented with 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin at 37°C in a 5% saturated CO2 incubator. When cells reached 80% confluency, they were harvested, centrifuged at 300 g for 10 minutes, and plated at 1200 cells / well in a 96-well plate. After 24 hours, PARPi was added at various final concentrations (0, 0.01, 0.1, 1, 10, 100, and 1000 nM) and cultured for an additional 72 hours. The medium was then replaced (with the same final concentration of PARPi added) and cultured for an additional 96 hours. The 96-well plate was removed and the OD values ​​at a wavelength of 450 nM were measured using the CCK8 assay. The cell inhibition rate was calculated as follows: % inhibition = 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%.

[0278] BRCA wild-type cells DLD-1 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum, 100U / mL penicillin, and 100μg / mL streptomycin, and cultured in a 5% saturated CO2 incubator at 37°C. When the cells grew to 80% confluence, the cells were collected, centrifuged at 300g for 10 minutes, and plated on a 96-well plate at 1000 cells / well. After 24 hours, different final concentrations of PARPi (0, 1, 10μM) were added and cultured for 72 hours. The cells were treated with a new medium (PARPi was re-added with the same final concentration) and cultured for 96 hours. The 96-well plate was removed, and the OD value at a wavelength of 450nM was detected by CCK8, and the cell inhibition rate was calculated:

[0279] Inhibition rate %=1-(average OD value of the drug administration group-average OD value of the Blank group) / (average OD value of the Control group-average OD value of the Blank group)*100%.

[0280] The test results are shown in Table 3 below:

[0281] Table 3 Cell antiproliferative activity test results

[0282] Conclusion: The compounds of the present invention have a significant inhibitory effect on BRCA mutant MDA-MB-436 cells, but have no significant inhibitory effect on BRCA wild-type DLD-1 cells, indicating that the compounds of the present invention specifically inhibit tumor cells with homologous recombination defects.

[0283] 3. Pharmacokinetic evaluation of the compound in Balb / c mice

[0284] Experimental purpose: To understand the pharmacokinetics of the compound.

[0285] Experimental basis: Technical Guidelines for Nonclinical Pharmacokinetic Studies of Chemical Drugs, 2014.

[0286] Experimental plan: The pharmacokinetics of the compound were investigated by intravenous administration (1 mg·kg-1) and oral administration (1 mg·kg-1) to Balb / c mice.

[0287] Sample preparation: weigh about 0.2 mg of the compound, add 10 μL of DMSO to dissolve it, and then add sodium chloride solution for injection to make a 0.1 mg·mL-1 compound solution for administration.

[0288] Sample collection: 6 male Balb / c mice (Chengdu Dashuo Experimental Animal Co., Ltd., license number: SCXK (Chuan) 2020-030), 3 were intravenously administered (IV) at 1 mg·kg-1, and 3 were gavage administered (PO) at 1 mg·kg-1. Approximately 0.05 mL of blood was collected 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, the supernatant plasma was collected, and frozen at -40 ° C for testing. The plasma concentration was quantitatively analyzed by LC-MS / MS analysis, and pharmacokinetic parameters such as peak time (Cmax), area under the concentration-time curve (AUC(0-t)), and half-life (T) were calculated. 1 / 2 ), clearance (CL), tissue distribution (Vdss), bioavailability (F), etc.

[0289] The results of the pharmacokinetic evaluation are shown in Table 4 below:

[0290] Table 4 Pharmacokinetic test results of the compounds in Balb / c mice

[0291] Conclusion: The compound of the present invention has good pharmacokinetic properties in Balb / c mice, including good oral bioavailability, exposure, half-life and clearance. max It is superior to reference compound 5305, compound 1, compound 2, compound 3, compound 15 and compound 21, etc.

[0292] 4. Pharmacokinetic evaluation of the compound in SD rats

[0293] Experimental purpose: To understand the pharmacokinetics of the compound.

[0294] Experimental basis: Technical Guidelines for Nonclinical Pharmacokinetic Studies of Chemical Drugs, 2014.

[0295] Experimental plan: The pharmacokinetics of the compound were investigated by oral and intravenous administration in SD rats.

[0296] Experimental Procedure: Weigh the compound, add a small amount of DMSO, and then add sodium chloride solution for injection to prepare a solution for administration. Six male SD rats were orally administered. Approximately 0.1 mL of blood was collected at 5, 15, 30, 1, 2, 4, 6, 8, 10, and 24 hours after administration. The blood was centrifuged at 3500 rpm for 15 minutes, and the supernatant plasma was collected. 5 μL of plasma was transferred to an EP tube and protein was precipitated by adding 100 μL of acetonitrile containing 20 ng·ml⁻¹ of internal standard SAHA. The solution was vortexed for 30 seconds and centrifuged at 13000 rpm for 15 minutes. The supernatant was transferred to a vial for analysis. The standard curve range is 10 to 10,000 ng·ml⁻¹.

[0297] The results of the pharmacokinetic evaluation are shown in Table 5 below:

[0298] Table 5 Pharmacokinetic test results of the compounds in SD rats

[0299] Conclusion: The compound of the present invention has good pharmacokinetic properties in SD rats, including good oral bioavailability, exposure, half-life and clearance. max Better than reference compounds AZD5305, compound 1 and compound 21.

[0300] 5. In vivo pharmacodynamic study of compounds 1 and 6 on the subcutaneous transplanted tumor model of breast cancer MDA-MB-436 in nude mice

[0301] (1) Main instruments and equipment

[0302] CO2 cell culture incubator: Yamato brand, model IP610; biological safety cabinet: Suzhou Antai brand, model BSC-1304ⅡA2; normal temperature centrifuge: Thermo Scientific brand, model SORVALLST 16;

[0303] 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 refrigerator: American Thermo brand, model ULTS1651;

[0304] Water purifier: Millipore, USA, model F7PNO9748; vertical autoclave: Yamato, model DKN812C; 1 mL disposable sterile syringe: Shanghai Kindly Enterprise Development Group Co., Ltd.; 1 mL disposable sterile insulin syringe: BD; weight scale: Shanghai Hengping Instrument Factory, model JY2002; analytical balance: SARTORIUS, model BCE95I-1CEU.

[0305] Thermohygrometer: Wuqiang Thermohygrometer Manufacturing Center, measurement number Ji 30260102; small centrifuge: American Thermo brand, model SORVALL LEGEND MICRO 17 Centrifuge; mini mixer: Yeasen brand, model ES-VM25; metal bath: SCILOGEX brand, model SCL120-S.

[0306] (2) Main software and data processing systems

[0307] Graphpad Prism, version 6.0, Graphpad Software Ltd., was used to organize and quantify data and to create bar graphs.

[0308] (3) Experimental animals

[0309] Strain: NOD / SCID, Grade: SPF, Source: Beijing Weitonglihua Biotechnology Co., Ltd.

[0310] (4) Modeling method

[0311] Tumor cell culture and preparation: For routine tumor cell line passage and culture, the culture medium contains 10% fetal bovine serum, 100U / mL penicillin, and 100μg / mL streptomycin. When the cells grow to 80% confluence, collect the cells and centrifuge at 300g for 10 minutes. Wash 3 times with pre-cooled PBS, collect the cells by centrifugation at 300g for 10 minutes, resuspend the cells with PBS, count the cells with a hemocytometer, adjust the cell concentration, and pre-cool the cell suspension on ice after the cells are collected. Take 100μL of cell suspension and inject it subcutaneously in the dorsal armpit of the mouse. When the tumor volume reaches 200-300mm 3 Group treatment started at that time.

[0312] (5) Observation and testing indicators

[0313] General status observation: Observed animals: All surviving experimental animals planned for observation; Observation time: Twice a day; Observation content: including but not limited to general manifestations, behavioral status, eyes, mouth, nose and mouth, ears, hair, feces, urine, genitals and other toxic symptoms. If any abnormalities are found, a detailed description is required.

[0314] (6)Weight

[0315] All surviving experimental animals planned for measurement were weighed before modeling for experimental grouping. After modeling, mice were weighed every two days, and weight changes were recorded in real time. Effectiveness criteria included tumor volume (TV), relative tumor volume (RTV), and relative tumor growth rate in the subcutaneous transplanted tumor model. The long and short diameters of the tumors were measured with a vernier caliper every two days to dynamically observe the antitumor effects 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.

[0316] Relative tumor volume (RTV) was calculated based on the measurement results using the formula: RTV = Vt / V0, where V0 is the TV measured at the time of dosing (d0) for each group, and Vt is the TV at each measurement for that group. The antitumor activity was evaluated using the relative tumor growth rate (T / C) (%), calculated as follows:

[0317] T / C(%)=TRTV / CRTV×100%;

[0318] TRTV: relative tumor volume of the treatment group; CRTV: relative tumor volume of the negative control group. Efficacy evaluation criteria: T / C (%) > 60 indicates ineffectiveness; T / C (%) ≤ 60, with a P < 0.05 as determined by statistical analysis, indicates efficacy. Tumor Weight Determination and Calculation of Tumor Inhibition Rate (%): At the end of treatment, animals were sacrificed, tumors were dissected, weighed, and photographed. The tumor inhibition rate (%) was calculated using the following formula:

[0319] Tumor inhibition rate (tumor growth inhibition rate, %) = (average tumor weight of negative control group (g) - average tumor weight of drug-treated group (g)) / average tumor weight of negative control group (g) × 100%;

[0320] Effectiveness criteria: Tumor inhibition rate (i.e., tumor growth inhibition rate) < 40% is considered ineffective; tumor inhibition rate ≥ 40% and P < 0.05 after statistical analysis is considered effective. Comprehensive criteria for subcutaneous tumor model effectiveness: Effectiveness is determined when either of the two effectiveness criteria (relative tumor growth rate and tumor inhibition rate) is met.

[0321] (7) Data collection and analysis

[0322] All raw data within the facility were collected manually or using a data acquisition system according to the experimental protocol and the relevant regulations of the research institution. Manually collected data can be transferred to software such as Excel for analysis and reporting. All data are quantitative data. Tables show the mean ± standard deviation (SD) for each animal group, and figures show the mean ± standard error (SEM). t-tests and survival analyses were performed using Excel and GraphPad Prism software.

[0323] Experimental Procedure: Forty-two female NOD / SCID mice were randomly divided into seven groups, each receiving a blank control, compound 6 (0.03, 0.1, and 0.3 mg / kg), the reference compound AZD5305 (0.1 mg / kg), compound 1 (0.1 mg / kg), and olaparib (100 mg / kg). All mice were orally administered once daily. Body weights were measured every two days, and tumor length and width were measured with a vernier caliper. After 24 days of administration, tumor-bearing mice were anesthetized and sacrificed. Tumor tissue was excised, weighed, and photographed for calculation of tumor inhibition rates.

[0324] Conclusion: The experimental results are shown in Figure 1 and Table 6. After cell inoculation, each drug-treated group was continuously administered for 24 days. Compared with the blank control group, each drug-treated 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) had a significant inhibitory effect on the growth of human breast cancer cell MM436 cell line NOD / SCID mouse subcutaneous tumor model. The tumor inhibition rates of each drug-treated group were 80.79%, 82.18%, 62.19%, 49.77%, 86.73%, and 95.83%, respectively; the relative tumor growth rate (T / C%) values ​​were 15.2%, 17.0%, 33.4%, 40.8%, 12.6%, and 3.8%, respectively. Among all the treatment groups, the compound 6 0.3 mg / kg group had the best tumor inhibition effect, with complete tumor regression in two mice (2 / 6) at the end of treatment. The compound 6 0.1 mg / kg group had a better tumor inhibition effect than the reference compounds AZD5305 and compound 1 at the same dose, and far better than the reference drug Olaparib 100 mg / kg group (P < 0.01). At the same time, there were significant dose-effect differences among the low, medium, and high dose groups of compound 6 (P < 0.001, P < 0.01).

[0325] Table 6 Tumor inhibition rate of compounds 1 and 6 on MDA-MB-436 nude mouse model for 24 days vs. blank group: c, P < 0.001

[0326] 6. Challenge experiment of compounds 1, 6 and 21 on the breast cancer MDA-MB-436 subcutaneous transplanted tumor model in nude mice

[0327] Experimental Procedure: Fifteen female NOD / SCID mice were randomly divided into three groups, each containing five mice: Compound 1 (1 mg / kg), Compound 6 (1 mg / kg), and Compound 21 (1 mg / kg). All mice were orally administered once daily. Body weights were measured every two days, and tumor length and width were measured using a vernier caliper.

[0328] Conclusion: The experimental results are shown in Figure 2. Compounds 1, 6, and 21 of the present invention still exhibited excellent antitumor efficacy in large tumor models, leading to complete tumor regression. Compound 6 exhibited a faster tumor regression rate than Compound 1 and Compound 21, indicating that at equivalent doses, Compound 6 exhibited stronger in vivo antitumor efficacy compared to Compound 1 and Compound 21.

[0329] 7. In vivo pharmacodynamic study of compound 6 in PDX gastric cancer mouse model

[0330] Implementation: A stable tumor-bearing mouse model was established using gastric cancer patient tissue. This study used tumor masses from mice that had been stably passaged three times for subcutaneous inoculation. Eight groups, each consisting of six mice, were divided into the negative control group (Control) and the carboplatin control group (Car) with 20 mg / kg, 0.2 ml / mouse, intraperitoneal injection once daily. The test drug, Compound 6, was administered orally at 0.3, 1, and 3 mg / kg, 0.2 ml / mouse, once daily. A combination group received Compound 6 plus carboplatin.

[0331] The specific dosing regimen and grouping are shown in Table 7.

[0332] Table 7 Experimental grouping table

[0333] Conclusion: The experimental results are shown in Figure 3. Different doses of compound 6 combined with carboplatin can inhibit the gastric cancer PDX model in a dose-dependent manner, especially the 3 mg / kg dose group of compound 6 combined with carboplatin, which significantly inhibited tumor growth.

[0334] 8. In vivo pharmacodynamic study of compound 6 on breast cancer SUM14PT subcutaneous transplanted tumor model in nude mice

[0335] Implementation: This study established a mouse model using subcutaneous inoculation of SUM14PT tumor cells. Eight groups, each containing six mice, were divided into the negative control group (control) and received normal saline (0.2 ml / mouse / dose) orally once daily. A carboplatin control group (Car) received 20 mg / kg (0.2 ml / mouse / dose) intraperitoneally once every seven days. The olaparib control group received 100 mg / kg (0.2 ml / mouse / dose) orally once daily. The test drug, compound 6, was administered orally at 0.3 and 1 mg / kg (0.2 ml / mouse / dose) orally once daily. Combination groups received either olaparib or compound 6 plus carboplatin.

[0336] The specific dosing regimen and grouping are shown in Table 8.

[0337] Table 8 Experimental grouping table

[0338] Conclusion: The experimental results are shown in Figure 4. Compound 6 0.3 and 1 mg / kg once daily orally administered alone had superior antitumor effects compared to olaparib 100 mg / kg once daily administered alone. Compound 6 1 mg / kg once daily orally administered in combination with carboplatin had superior efficacy compared to olaparib 100 mg / kg once daily orally administered in combination with carboplatin.

Claims

1. A compound of formula II or a pharmaceutically acceptable form thereof, characterized in that: The structure of Formula II is as follows: in: Indicates a single bond or a double bond; R1 is selected from C 1-4 deuterated alkyl; 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 only one selected from N; R 2a and R 2b independently selected from hydrogen, deuterium, methyl or deuterated methyl; R3 is selected from deuterium, fluorine, C 1-4 Alkyl or C 1-4 at least one of a deuterated alkyl group; R 3a Selected from hydrogen, deuterium, fluorine, hydroxyl, cyano, C 1-4 Alkyl, C 1-4 Fluorinated alkyl or C 1-4 alkoxy; R4 is selected from hydrogen, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluoroalkyl or -CONHR7; R 5a 、R 5a and R 5c independently selected from hydrogen, fluorine, chlorine, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluorinated alkyl, C 1-4 Alkoxy or C 1-4 Fluoroalkoxy; R6 is selected from hydrogen, fluorine, chlorine, C 1-4 Alkyl, C 1-4 Fluorinated alkyl or C 1-4 deuterated alkyl; R7 is selected from hydrogen, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluorinated alkyl or 3-6 membered cycloalkyl; 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 ); R 9a 、R 9b 、R 9c and R 9d independently selected from hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluorinated alkyl, C 1-4 Alkoxy or C 1-4 Fluoroalkoxy; n1 is an integer from 0 to 8; n3 is independently selected from 0 or 1; The pharmaceutically acceptable form is selected from pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, N-oxides, isotopically labeled substances, metabolites, or prodrugs.

2. The compound according to claim 1, characterized in that: R1 is selected from deuterated methyl or deuterated ethyl.

3. The compound according to claim 2, characterized in that: R1 is selected from -CD2CD3 or -CH2CD3.

4. The compound according to any one of claims 1 to 3, characterized in that: R3 is at least one selected from deuterium, methyl, deuterated methyl, fluoromethyl or methoxy.

5. The compound according to claim 4, characterized in that: R3 is selected from deuterium or methyl.

6. The compound according to any one of claims 1 to 5, characterized in that: When it is a double bond, R 3a does not exist, When it is a single bond, R 3a Selected from hydrogen and deuterium.

7. The compound according to any one of claims 1 to 6, characterized in that: R4 is selected from fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluorinated alkyl, C 1-4 Alkyl, C 1-4 Deuterated alkylaminoacyl, C 1-4 Fluoroalkylaminoacyl or 3-6 membered cycloalkylaminoacyl; Preferably, R4 is selected from fluorine, cyano, methylaminoacyl, deuterated methylaminoacyl or cyclopropylaminoacyl.

8. The compound according to any one of claims 1 to 7, characterized in that: R6 is selected from hydrogen, fluorine, chlorine or methyl.

9. The compound according to any one of claims 1 to 8, characterized in that: R 5a 、R 5b and R 5c are independently selected from hydrogen, fluorine, chlorine or methyl.

10. The compound according to any one of claims 1 to 9, characterized in that: It has the structure described in formula VI-1: in: R6 is selected from hydrogen or fluorine; R 9a Selected from hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Fluorinated alkyl; n1 is an integer from 0 to 6.

11. The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable form thereof, characterized in that: It has the structure described in formula VI-3: in: R6 is selected from hydrogen or fluorine; R 9a Selected from hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Fluorinated alkyl; n1 is an integer from 0 to 6.

12. The compound according to any one of claims 1 to 11, characterized in that: The compound is selected from:

13. The compound according to any one of claims 1 to 11, characterized in that: The compound is selected from:

14. The compound according to any one of claims 1 to 11, characterized in that: The compound is selected from:

15. A pharmaceutical composition, characterized in that: The active ingredient is a compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite or prodrug thereof, supplemented with a pharmaceutically acceptable carrier.

16. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, and the pharmaceutical composition according to claim 15, are used in the preparation of a medicament for preventing and / or treating PARP1 enzyme-related diseases.

17. The use according to claim 16, characterized in that: The PARP1 enzyme-related disease is a tumor-related disease.

18. The use according to claim 17, characterized in that: The tumor-like disorders are deficient in the HR-dependent DNA DSB repair pathway.

19. The use according to claim 17 or 18, characterized in that: The neoplastic disorder comprises one or more cancer cells that have a reduced or absent ability to repair DNA DSBs by HR relative to normal cells.

20. The use according to claim 19, characterized in that: The cancer cells have a BRCA1 or BRCA2 deficient phenotype.

21. The use according to any one of claims 17 to 20, characterized in that: The tumor-related disease is breast cancer, ovarian cancer, primary peritoneal cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer, glioblastoma or lung cancer.

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