Fused heterocyclic compounds and their uses

Abstract

The present invention discloses a series of fused heterocyclic compounds and their uses, which belong to the technical field of chemical medicine.The fused heterocyclic compounds represented by Formula II and provided by the present invention can function as PARP1 inhibitors, and have the advantages of high activity and high selectivity, as well as excellent pharmacokinetic properties and excellent safety. [Formula 1] TIFF2025535816000053.tif31170

Description

[Technical Field]

[0001] The present invention belongs to the field of chemical medicine and relates to a series of fused heterocyclic compounds and their uses. [Background technology]

[0002] During cell growth, DNA is constantly damaged by various internal and external factors. The most serious types of DNA damage are single-strand breaks (SSBs) and double-strand breaks (DSBs), with SSBs being more common. If these breaks are not repaired in a timely and accurate manner, they can lead to genomic instability, which can subsequently cause carcinogenesis or directly result in cell death. For single-strand breaks in DNA, repair relies primarily on the PARP enzyme. For double-strand breaks in double-stranded DNA, there are two main repair pathways: non-homologous end joining repair and homologous recombination (HR) repair. Homologous recombination repair is a high-fidelity, error-free repair mechanism and the primary pathway for double-stranded DNA repair. Homologous recombination repair involves numerous proteins, the most well-known of which is the BRCA protein. Two studies in 2005 (Non-Patent Document 1 and Non-Patent Document 2) demonstrated that tumor cells lacking BRCA1 or BRCA2 were selectively inhibited by PARP inhibitors. Based on these findings, scholars proposed the concept of synthetic lethality: the loss of either the BRCA or PARP gene is not lethal in itself, but the simultaneous inactivation of both leads to cell death. Based on the synthetic lethality theory, PARP inhibitors (PARPi) were developed to selectively target BRCA1 / 2 mutant cancer cells.

[0003] PARP inhibitors have demonstrated excellent clinical efficacy in patients with homologous recombination repair-deficient cancers. However, whether administered as monotherapy or in combination therapy, hematological toxicity (anemia, neutropenia, and thrombocytopenia) and other toxicities limit the application of these drugs. Related research (Non-Patent Document 3) has shown that these adverse reactions may be due to the inhibition of PARP2 by commercially available PARP inhibitors, even though PARP2 is not essential for therapeutic efficacy. Highly selective PARP1 inhibitors can reduce hematological toxicity, enhance the therapeutic safety margin, and increase the possibility of combination with other chemotherapy drugs or targeted therapies.

[0004] Therefore, there remains an unmet clinical need for effective and safe PARP inhibitors, especially those that are selective for PARP 1. The novel PARP1 inhibitors described in the present invention exhibit significantly higher selectivity for PARP 1 compared to other PARP family members (e.g., PARP2, PARP3, PARP5a, and PARP6) and can be used to treat diseases associated with PARP function. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] 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 [Non-patent document 2] 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) [Non-patent document 3] Harris PA,Boloor A,Cheung M,et al.Discovery of 5-[[4-[(2,3-dimethyl-2H-indazol-6-yl)methylam-ino]-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 Summary of the Invention [Problem to be solved by the invention]

[0006] An object of the present invention is to provide certain fused heterocyclic compounds and their use for achieving highly selective and effective prevention or treatment of diseases related to PARP function. [Means for solving the problem]

[0007] In a first aspect, the present invention provides a compound represented by Formula II or a pharmaceutically acceptable form thereof, wherein Formula II is as follows: [ka] During the ceremony, [ka] represents a single bond or a double bond, R1 is C 1-4 deuterated alkyl; X1 is N or C(R 5a ) and X2 is selected from N or C(R 5b ) and X3 is selected from N or C(R 5c), and exactly one of X1, X2, and X3 is selected from N; R 2a and R 2b are independently selected from hydrogen, deuterium, methyl, or deuteromethyl; R3 is deuterium, fluorine, C 1-4 Alkyl or C 1-4 deuterated alkyl; R 3a is hydrogen, deuterium, fluorine, hydroxy, cyano, C 1-4 Alkyl, C 1-4 Fluoroalkyl or C 1-4 alkoxy; R4 is 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 are hydrogen, fluorine, chlorine, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluoroalkyl, C 1-4 Alkoxy or C 1-4 fluoroalkoxy; R6 is hydrogen, fluorine, chlorine, C 1-4 Alkyl, C 1-4 Fluoroalkyl or C 1-4 deuterated alkyl; R7 is hydrogen, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 selected from fluoroalkyl or 3- to 6-membered cycloalkyl; X5 is nitrogen or C(R 9a ) and X6 is selected from nitrogen or C(R 9b ) and X7 is selected from nitrogen or C(R 9c ) and X8 is selected from nitrogen or C(R 9d ) and R 9a , R 9b , R 9cand R 9d is hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluoroalkyl, C 1-4 Alkoxy or C 1-4 fluoroalkoxy; n1 is an integer selected from 0 to 8, n3 is independently selected from 0 or 1.

[0008] The pharmaceutically acceptable form is selected from the group consisting of a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug.

[0009] In some embodiments of the present invention, for compounds represented by Formula II above or a pharmaceutically acceptable form thereof, R 1 is selected from deuterated methyl or deuterated ethyl.

[0010] In some preferred embodiments of the present invention, for compounds represented by Formula II above or a pharmaceutically acceptable form thereof, R1 is selected from -CD2CD3 or -CH2CD3.

[0011] In some embodiments of the present invention, for compounds represented by Formula II above or a pharmaceutically acceptable form thereof, R3 is selected from deuterium, methyl, deuteriummethyl, fluoromethyl, or methoxy.

[0012] In some preferred embodiments of the present invention, R3 of the compounds represented by Formula II above, or a pharmaceutically acceptable form thereof, is selected from deuterium or methyl.

[0013] In some embodiments of the present invention, for the compound represented by Formula II above or a pharmaceutically acceptable form thereof: [ka] When is a double bond, R 3a does not exist, [ka] When is a single bond, R 3a is selected from hydrogen and deuterium.

[0014] In some embodiments of the present invention, for compounds represented by Formula II above or a pharmaceutically acceptable form thereof, R4 is fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 1-4 Fluoroalkyl, C 1-4 Alkylaminocarbonyl, C 1-4 Deuterated alkylaminocarbonyl, C 1-4 It is selected from fluoroalkylaminocarbonyl and 3- to 6-membered cycloalkylaminocarbonyl.

[0015] In some preferred embodiments of the present invention, for compounds represented by Formula II above or a pharmaceutically acceptable form thereof, R4 is selected from fluoro, cyano, methylaminocarbonyl, deuterated methylaminocarbonyl, and cyclopropylaminocarbonyl.

[0016] In some preferred embodiments of the present invention, for compounds represented by Formula II above or a pharmaceutically acceptable form thereof, R6 is selected from hydrogen, fluorine, chlorine, or methyl.

[0017] In some embodiments of the present invention, for compounds represented by Formula II above or a pharmaceutically acceptable form thereof, R 5a , R 5b and R 5c are independently selected from hydrogen, fluorine, chlorine or methyl.

[0018] In some embodiments of the present invention, in addition to the compounds represented by Formula II or a pharmaceutically acceptable form thereof, the present invention also provides compounds having a structure represented by Formula VI-1: [ka] During the ceremony, R6 is selected from hydrogen or fluorine; R 9a is hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 fluoroalkyl; n1 is an integer selected from 0 to 6.

[0019] In some embodiments of the present invention, in addition to the compound represented by Formula II or a pharmaceutically acceptable form thereof, the present invention also provides a compound having a structure represented by Formula VI-3: [ka] During the ceremony, R6 is selected from hydrogen or fluorine; R 9a is hydrogen, fluorine, chlorine, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 fluoroalkyl; n1 is an integer selected from 0 to 6.

[0020] In some more preferred embodiments of the present invention, the following compounds, or pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites, or prodrugs thereof, are provided: [ka]

[0021] In some more preferred embodiments of the present invention, the following compounds, or pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites, or prodrugs thereof, are provided: [ka]

[0022] In some more preferred embodiments of the present invention, the following compounds, or pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites, or prodrugs thereof, are provided: [ka]

[0023] In a second aspect, the present invention provides a pharmaceutical composition comprising, as an active ingredient, a compound as described above (Formula II, Formula VI-1 or Formula VI-3) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug, and a pharmaceutically acceptable carrier.

[0024] A further object of the present invention is to provide a process 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 salt thereof, or a mixture thereof, with one or more pharmaceutically acceptable carriers.

[0025] Pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions of the present invention are pharmaceutically acceptable carriers, and examples of suitable pharmaceutically acceptable carriers are as described in Remington's Pharmaceutical Sciences (2005).

[0026] The pharmaceutical composition can be administered in any form as long as it achieves the prevention, alleviation, suppression, or cure of symptoms in a human or animal patient. For example, various suitable dosage forms can be prepared according to the administration route.

[0027] In other embodiments, administration of the compounds or pharmaceutical compositions of the present invention can be combined with another treatment modality, which can be selected from, but is not limited to, radiation therapy, chemotherapy, immunotherapy, or a combination thereof.

[0028] The present invention further relates to a pharmaceutical formulation comprising any compound represented by 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 semi-solid formulation, a liquid formulation, or a gaseous formulation.

[0029] A further object of the present invention is to provide a product, for example, in the form of a kit. The products referred to herein include, but are not limited to, kits and packages. The product of the present invention includes (a) a first container, (b) a pharmaceutical composition disposed in the first container, the composition being any compound represented by Formula II, Formula VI-1, or Formula VI-3, or a pharmaceutically acceptable form thereof, or a mixture thereof, (c) optionally, a packaging design indicating that the pharmaceutical composition can be used to treat a neoplastic disorder (defined below), and (d) a second container.

[0030] The first container is a container for containing the pharmaceutical composition. This container can be used for preparation, storage, shipping, and / or individual / bulk sales. The first container is intended to encompass bottles, cans, vials, flasks, syringes, tubes (e.g., for cream products), or any other container for preparing, containing, storing, or dispensing a pharmaceutical product.

[0031] The second container is a container for housing the first container and, optionally, a packaging specification. Examples of the second container include, but are not limited to, a box (such as a cardboard box or plastic box), a crate, a carton, a bag (such as a paper bag or plastic bag), a pouch, and a burlap sack. The packaging specification may be physically attached to the exterior of the first container via ties, adhesive, staples, or another adhesive means, or may be disposed inside the second container without requiring any physical attachment to the first container. Alternatively, the packaging specification is disposed on the exterior of the second container. When disposed on the exterior of the second container, it is preferable that the packaging specification be physically attached via ties, adhesive, staples, or another adhesive means. Alternatively, the packaging specification may abut or contact the exterior of the second container without requiring physical attachment.

[0032] The packaging specification may include a trademark, label, marking, etc., that lists information related to the pharmaceutical composition disposed within the first container. The listed information is typically determined by a regulatory agency (e.g., the U.S. Food and Drug Administration) that governs the geographic area in which the product is intended for sale. Preferably, the packaging specification specifically lists the labeling for which the pharmaceutical composition is approved. The packaging specification may be made of any material from which the information contained therein or thereon can be read. Preferably, the packaging specification is a printable material (paper, plastic, cardboard, foil, adhesive paper, plastic, etc.) onto which the required information can be formed (e.g., printed or applied).

[0033] In a third aspect, the present invention provides the use of the above-mentioned compounds, compounds of formula II, VI-1 or VI-3, as well as related specific compounds or pharmaceutically acceptable forms thereof, or pharmaceutical compositions of the invention, in the manufacture of a medicament for preventing or treating a PARP1 enzyme-related disease.

[0034] The present invention provides a method for preventing or treating a PARP1 enzyme-associated disease, comprising administering to an individual in need thereof a compound represented by formula II, formula VI-1 or formula VI-3, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention.

[0035] The present invention provides a compound represented by Formula II, Formula VI-1 or Formula VI-3, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention, for preventing or treating a PARP1 enzyme-related disease.

[0036] The present invention provides a method for preventing or treating a PARP1 enzyme-related disease using a compound represented by Formula II, Formula VI-1, or Formula VI-3, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention, in combination with an additional therapeutic method, including, but not limited to, radiation therapy, chemotherapy, immunotherapy, or a combination thereof.

[0037] In some embodiments, the PARP1 enzyme-associated disease is a disease that is sensitive or responsive to PARP1 enzyme inhibition.

[0038] In some embodiments, the PARP1 enzyme-associated disease is a neoplastic disorder.

[0039] In some preferred embodiments, the neoplastic disorder is deficient in the HR-dependent DNA DSB repair pathway.

[0040] In some preferred embodiments, the neoplastic disorder comprises one or more types of cancer cells that have a reduced or absent ability to repair DNA DSBs by HR compared to normal cells.

[0041] In some preferred embodiments, the cancer cells have a BRCA1 or BRCA2 deficient phenotype.

[0042] In some embodiments, the PARP1 enzyme-associated disease is an oncological disorder, including, but not limited to, solid and hematological malignancies. In further embodiments, oncological disorders include, but are not limited to, breast cancer, colorectal cancer, colon cancer, lung cancer (including small cell lung cancer, non-small cell lung cancer, and bronchoalveolar carcinoma), and prostate cancer, as well as bile duct cancer, bone cancer, bladder cancer, head and neck cancer, kidney cancer, liver cancer, cancer of the gastrointestinal tissue, 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).

[0043] In some preferred embodiments, the PARP1 enzyme-related disease is breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, hematological cancer, gastrointestinal cancer, or lung cancer.

[0044] In a further preferred embodiment, the compounds of the present invention can be used in combination with radiation therapy and chemotherapy or immunotherapy for the prevention or treatment of cancer.

[0045] The beneficial effects of the present invention are as follows:

[0046] The present invention provides a novel class of highly active and highly selective PARP1 inhibitors that can achieve at least one of the following technical effects: (1) high inhibitory activity against the PARP1 enzyme; (2) selective inhibition of PARP1 with high selectivity for other enzymes in the PARP family, such as PARP2, PARP5a, and PARP5b; (3) strong inhibitory activity against homologous recombination repair-deficient tumor cells and weak inhibitory effect in non-homologous recombination repair-deficient cells; (4) excellent pharmacokinetic properties (good bioavailability, suitable half-life, and duration of action); and (5) excellent safety (reduced toxicity and / or reduced side effects, and a wide therapeutic window).

[0047] Definition of Terms

[0048] Unless defined herein below, all technical and scientific terms used herein are intended to have the meanings commonly understood by those of ordinary skill in the art. As used herein, "including," "comprising," "having," "containing," or "relating to," and other variations thereof, are inclusive or open-ended and do not exclude other, unlisted elements or method steps. Those skilled in the art will understand that the above terms, such as "including," encompass the meaning of "consisting of."

[0049] As used herein, the terms "one," "a," "the," "at least one," and "one or more" can be used interchangeably. Thus, for example, a composition comprising "a" pharmaceutically acceptable excipient can be interpreted to mean that the composition comprises "one or more" pharmaceutically acceptable excipients.

[0050] For example, "C 1-4 " is C 2-4 , C 3-4 , C 1-2 , C 1-3 , C 1-4 etc., in addition to C1, C2, C3, C4, etc., any subranges and point values ​​therein should be understood to encompass these.

[0051] Unless otherwise stated, as used herein [ka] represents a single bond or a double bond.

[0052] In the present invention, unless otherwise stated, halogen refers to fluorine, chlorine, bromine or iodine.

[0053] As used herein, unless otherwise specified, the term "alkyl" includes linear or branched monovalent saturated hydrocarbon groups. For example, alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 3-(2-methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl, 2-methylpentyl, and the like. Similarly, "C 1-4 C in "Alkyl" 1-4 refers to groups having 1, 2, 3 or 4 carbon atoms arranged in a straight or branched chain configuration.

[0054] In the present invention, unless otherwise specified, "cycloalkyl," "carbocyclic," or "cycloalkylene" refers to a saturated or partially saturated monocyclic or polycyclic (including bicyclic) non-aromatic hydrocarbon group. Typical cycloalkyl groups include, but are not limited to, monocyclic cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, and the like, or bicyclic cycloalkyl groups containing fused, bridged, 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, decahydronaphthyl, and the like. For example, "C 3-12 The term "cycloalkyl" refers to a cycloalkyl group having 3 to 12 ring carbon atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12). Cycloalkyl or cycloalkylene groups of the present invention are optionally substituted with one or more substituents described herein.

[0055] In the present invention, unless otherwise specified, the term "fluoroalkyl" refers to an alkyl group as defined above, in which one or more hydrogen atoms are replaced by a fluorine atom. For example, "C 1-4 The term "fluoroalkyl" refers to a C alkyl group optionally substituted with one or more (1-3) fluorine atoms. 1-4It refers to an alkyl group. Those skilled in the art will understand that when there are two or more fluorine atom substituents, the fluorine atoms can be the same or different and can be located on the same or different C atoms. Examples of haloalkyl groups include, but are not limited to, -CHF, -CHF, -CF, -CF, -CHCF, -CHCHCF, etc. The fluoroalkyl groups of the present invention are optionally substituted with one or more substituents as described herein.

[0056] The present invention also encompasses all pharmaceutically acceptable isotopically labeled compounds that are identical to the compounds of the present invention except that one or more atoms are replaced with an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or atomic number predominant in nature. Examples of isotopes that are suitable for incorporation into compounds of the present invention include isotopes of hydrogen (e.g., deuterium ( 2 H) and tritium ( 3 H), isotopes of carbon (e.g., 13 C and 14 C), isotopes of chlorine (e.g., 37 Cl), isotopes of iodine (e.g., 125 I), isotopes of nitrogen (e.g., 13 N and 15 N), isotopes of oxygen (e.g., 17 O and 18 O), isotopes of phosphorus (e.g., 32 P) and sulfur isotopes (e.g., 34 S), but are not limited to these.

[0057] In the present invention, the term "polymorph" refers to different solid crystalline phases of a particular compound of the present invention resulting from the existence of two or more different molecular arrangements in the solid state. A particular compound of the present invention may exist in more than one crystalline form, and the present invention is intended to encompass all such crystalline forms and mixtures thereof. Generally, crystallization results in the formation of a solvate of the compound of the present invention. As used herein, the term "solvate" refers to an aggregate containing 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. While the compounds of the present invention may form true solvates, in some cases they may simply retain non-stoichiometric water or mixtures of water in combination with a portion of the non-stoichiometric solvent. The compounds of the present invention may be reacted in a solvent or precipitated or crystallized from a solvent. Solvates of the compounds of the invention are also included within the scope of the invention. The invention also includes all possible crystalline forms or polymorphs of the compounds of the invention, including single polymorphs or mixtures of two or more polymorphs in any ratio.

[0058] In the present invention, the term "stereoisomer" refers to an isomer resulting from the presence of at least one asymmetric center. Compounds with one or more (e.g., one, two, three, or four) asymmetric centers may give rise to racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers. Certain individual molecules may also exist as geometric isomers (cis / trans). Similarly, compounds of the present invention may exist as mixtures of two or more structurally distinct forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, and imine-enamine tautomers. It is to be understood that the scope of the present invention encompasses all such isomers or mixtures of isomers in any ratio (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% and 99%).

[0059] 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 summary 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. The term "pharmaceutically acceptable acid addition salt" refers to a salt formed with an inorganic or organic acid that retains the bioavailability of the free base without other adverse effects. Inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc., and organic acid salts include, but are not limited to, formate, acetate, 2,2-dichloroacetate, trifluoroacetate, propionate, caproate, caprylate, caprate, 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, naphthalenedisulfonate, etc. These salts may be prepared by methods known in the patent. The term "pharmaceutically acceptable base addition salt" refers to salts formed with inorganic or organic bases that retain the biological effectiveness of the free acids without other adverse 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, etc. 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 amines, secondary amines, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Preferred organic bases include, but are not limited to, isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and hexocaffeine. These salts may be prepared by methods known in the patent.

[0060] In the present invention, unless otherwise specified, the term "ester" refers to an ester derived from a compound described herein, including a physiologically hydrolyzable ester (which can be hydrolyzed under physiological conditions to release a compound of the invention in the free acid or alcohol form). The compound of the invention can also be an ester itself.

[0061] The compounds of the present invention may exist in the form of solvates (preferably hydrates), which contain polar solvents, especially water, methanol, ethanol, etc., as structural elements of the crystalline lattice of the compounds. The amount of polar solvent, especially water, may be present in a stoichiometric or non-stoichiometric ratio. Those skilled in the art will understand that not all nitrogen heterocycles are capable of forming N-oxides, since the nitrogen requires an available lone pair of electrons to be oxidized to an oxide. Those skilled in the art will recognize nitrogen heterocycles that are capable of forming N-oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include the oxidation of heterocycles and tertiary amines using peroxyacids such as peroxyacetic acid and m-chloroperoxybenzoic acid (mCPBA), hydrogen peroxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, and dioxiranes such as sodium perborate and dimethyldioxirane. These methods for preparing N-oxides have been thoroughly described and reviewed in the literature. See, for example, T.L.G. Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp. 748-750 (A.R.Katritzky and A.J.Boulton, Eds., Academic Press); and G.W.H. Cheeseman and E.S.G. Werstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp. 390-392 (A.R.Katritzky and A.J.Boulton, Eds., Academic Press).

[0062] As used herein, the term "metabolite" refers to a substance formed in the body when a compound of the invention is administered. Metabolites of a compound may be identified using techniques well established in the art and characterized by experimental methods. For example, such products may result from oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic degradation, and other reactions of the administered compound. Accordingly, the invention encompasses metabolites of the compounds of the invention, including compounds produced by a process comprising contacting a compound of the invention with a mammal for a period of time sufficient to yield a metabolic product thereof.

[0063] In the present invention, the term "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 when administered into or onto the body, for example, by hydrolytic cleavage. Typically, such prodrugs are functional derivatives of compounds that can be easily converted into the desired therapeutically active compound in vivo. Further information on the use of prodrugs can be found in "Prodrugs as Novel Delivery Systems," Volume 14, ACS Symposium Series (T. Higuchi and V. Stella). For example, prodrugs of the present invention can be prepared by replacing appropriate functional groups present in the compounds of the present invention with certain moieties known to those skilled in the art as "promoieties" (e.g., as described in "Design of Prodrugs" by H. Bundgaard, Elsevier, 1985).

[0064] As used herein, the term "pharmaceutical composition" refers to a formulation of a compound of the present invention containing a medium generally accepted in the art for delivery of a biologically active compound to a mammal, such as a human. The medium comprises a pharmaceutically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate administration to an organism and promote absorption of the active ingredient, thereby allowing the compound to exert its biological activity.

[0065] As used herein, the term "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier approved by the relevant government regulatory agency or approved for use in humans or veterinary animals.

[0066] As used herein, the terms "drug combination," "co-drug administration," "combination therapy," "administration of another therapy," "administration of another therapeutic agent," and the like refer to a therapeutic regimen obtained by mixing or combining two or more active ingredients, including both fixed and non-fixed combinations of active ingredients. The term "fixed combination" refers to the simultaneous administration to a patient of at least one compound described herein and at least one synergist in the form of a single entity or dosage form. The term "non-fixed combination" refers to the simultaneous, concurrent, or sequential administration to a patient of at least one compound described herein and at least one synergist as separate entities, with adjustable intervals between administrations. These principles also apply to cocktail therapy, such as the administration of three or more active ingredients.

[0067] In the present invention, unless otherwise specified, the term "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 carcinoma, lung adenocarcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, cervical cancer, ovarian cancer, colon cancer, nasopharyngeal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, oral cancer, and other diseases.

[0068] In the present invention, unless otherwise stated, the term "treatment" refers to reversing, alleviating, inhibiting the progression of, or one or more symptoms of, the disorder or condition to which such term applies, or preventing such disorder or condition, or one or more symptoms thereof.

[0069] Without departing from the common general knowledge in the art, the preferred technical features described above can be freely combined in any way, thereby resulting in various preferred embodiments of the present invention. [Brief explanation of the drawings]

[0070] [Figure 1] 1 shows the change in tumor volume in an MDA-MB-436 nude mouse model after administration of compounds 1 and 6. [Figure 2]1 shows the changes in tumor volume in an MDA-MB-436 nude mouse model after administration of compounds 1, 6, and 21. [Figure 3] This shows the change in tumor volume in gastric cancer PDX (patient-derived xenograft) model mice after combination therapy with compound 6 and carboplatin. [Figure 4] 1 shows the change in tumor volume in a SUM14PT nude mouse subcutaneous xenograft model after combination therapy with Compound 6 and carboplatin. DETAILED DESCRIPTION OF THE INVENTION

[0071] Detailed Description of the Invention

[0072] The scheme of the present invention will be described below in conjunction with the embodiments. Those skilled in the art will understand that the following embodiments are intended to be only illustrative of the present invention and should not be considered as limiting the scope of the present invention. If specific techniques or conditions are not described in the embodiments, the techniques or conditions described in the technical literature or product specifications shall prevail. [Example]

[0073] Reagents and materials used in embodiments of the present invention are commercially available.

[0074] [Table 1]

[0075] The structures of the compounds described in this invention are determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were carried out using a Bruker AVANCE-400 spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d), deuterated chloroform (CDCl) and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard. Chemical shifts were measured at 10-6 It is reported in parts per million (ppm).

[0076] Mass spectrometry (MS) was performed using an Agilent SQD(ESI) mass spectrometer (manufacturer: Agilent, signal: 6110).

[0077] HPLC analysis was performed using an Agilent 1200DAD high pressure liquid chromatograph (SunFire C18, 150 x 4.6 mm, 5 wn, column) and a Waters 2695-2996 high pressure liquid chromatograph (Gemini C18, 150 x 4.5 mm, 5 ym, column).

[0078] Thin-layer chromatography (TLC) was performed using Qingdao Haiyang GF254 silica gel plates. For analytical TLC, plates with a thickness of 0.15–0.2 mm were used, whereas for preparative TLC, silica gel plates with a thickness of 0.4–0.5 mm were used.

[0079] Column chromatography generally uses Qingdao Haiyang 100-200 or 200-300 mesh silica gel as the carrier.

[0080] Unless otherwise noted in the following embodiments, all reactions are carried out under an argon or nitrogen atmosphere. An argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen balloon having a volume of about 1 L. A hydrogen atmosphere refers to a reaction flask connected to a hydrogen balloon having a volume of about 1 L. Hydrogenation reactions are typically carried out by evacuating, filling with hydrogen, and repeating this process three times.

[0081] Preparation of intermediates

[0082] Intermediate int-1: N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide [ka]

[0083] Step 1: Compound 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 mixture of 7 mL of dioxane, 3 mL of anhydrous ethyl alcohol, and 4 mL of water. The mixture was then purged with nitrogen three times and reacted at 90 °C under nitrogen protection for 2 h. After confirming completion of the reaction by TLC, the reaction mixture was cooled to room temperature, and 30 mL of dichloromethane and 20 mL of water were added. The layers were separated using a separatory funnel. The aqueous phase was extracted twice with dichloromethane, and the combined organic phases were washed sequentially with water and saturated brine. The reaction mixture was dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound int-1c (1 g, white solid).

[0084] Step 2: Compound Int-1c (1 g, 3 mmol), aqueous methylamine solution (5 g, 161.3 mmol), and anhydrous methanol (20 mL) were added to a 100 mL reaction flask, and the mixture was stirred and reacted at room temperature overnight. After completion of the reaction was confirmed by TLC, the reaction mixture was concentrated to dryness under reduced pressure to give compound Int-1d (0.8 g, white solid).

[0085] Step 3: Compound int-1d (0.5 g, 1.5 mmol) was added to 10 mL of anhydrous methanol, followed by 10 mL of a 4 mol / L solution of hydrochloric acid and dioxane. The mixture was stirred at room temperature for 0.5 to 1 h. After completion of the reaction was confirmed by TLC, the reaction mixture was concentrated under reduced pressure to give compound int-1 (0.5 g, white solid).

[0086] Embodiment 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 [ka]

[0087] Step 1: Compound 1a (20 g, 95.1 mmol) and selenium dioxide (16 g, 144 mmol) were added to a 250 mL reaction flask. 120 mL of 1,4-dioxane was then added. The mixture was heated to 110° C., and the reaction was stirred overnight. After TLC confirmed the completion of the reaction, the reaction liquid was filtered, and the filter residue was rinsed with ethyl acetate. The filtrates were combined and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 1b (16 g, yellow solid). LC-MS: ESI [M+H] + =225.2.

[0088] Step 2: Sodium hydride (6.86 g, 171.4 mmol) was added to a 250 mL reaction flask, followed by 60 mL of 1,4-dioxane. The mixture was then purged with nitrogen three times and cooled to 0° C. Compound 2-triethylphosphonobutanoate (43.2 g, 171.4 mmol) was slowly added dropwise under nitrogen protection. The reaction mixture was stirred at 0° C. for 10 minutes, then heated to room temperature and stirred for another 10 minutes, followed by heating at 40° C. and stirring for 5 minutes. The reaction mixture was cooled to −78° C. Compound INT1b (16 g, 71.4 mmol) was dissolved in 60 mL of 1,4-dioxane and then slowly added dropwise. The reaction mixture was stirred at −78° C. for 1 hour. After completion of the reaction was confirmed by TLC, the reaction mixture was quenched by slowly adding ice-cold saturated aqueous ammonium chloride solution. The mixture was extracted three times with 150 mL of ethyl acetate, and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 1c (13.26 g, brown liquid). LC-MS: ESI [M+H] + =323.3.

[0089] Step 3: Compound 1c (13.26 g, 41.1 mmol) was added to 100 mL of absolute ethyl alcohol, followed by the addition of Pd / C (1.33 g, 10%). The reaction mixture was stirred and allowed to react at room temperature overnight. After confirming the completion of the reaction by LC-MS, the reaction liquid was filtered, the filter residue was thoroughly rinsed with ethanol, and the filtrates were combined and concentrated by rotary evaporation. A 4 mol / L hydrochloric acid solution was added to 1,4-dioxane (50 mL), and the mixture was stirred at room temperature for 30 minutes. Ether was then added to precipitate a large amount of solid. The solid was filtered and dried to obtain compound 1d (7.32 g, white solid). LC-MS: ESI [M+H] + =249.3; 1 H NMR(400MHz,DMSO-d6)δ10.44(s,1H),8.62(d,J=1.9Hz,1H),7.75(d,J=1.9Hz,1H),4.34(q,J=7.1Hz,2H),3.87(s,0H),3.24(dd,J= 16.8,6.3Hz,1H),2.97(dd,J=16.8,10.1Hz,1H),1.81-1.65(m,1H),1.52-1.36(m,1H),1.32(t,J=7.1Hz,3H),0.93(t,J=7.4Hz,3H).

[0090] Step 4: Compound 1d (7.32 g, 29.5 mmol) was added to a 250 mL reaction flask, followed by 120 mL of 1,4-dioxane. DDQ (7.38 g, 32.5 mmol) was then added and refluxing was initiated. The mixture was allowed to react overnight. After confirming the completion of the reaction by LC-MS, the reaction liquid was concentrated by rotary evaporation. Saturated aqueous sodium bicarbonate solution was added and stirred for 1 hour. The liquid was filtered, and the filter residue was washed with water, followed by a small amount of ether. The residue was dried to give compound 1e (2.31 g, yellow solid). LC-MS: ESI [M+H] + =247.3.

[0091] Step 5: Compound 1e (2.0 g, 8.1 mmol) and 60 mL of tetrahydrofuran were added to a 150 mL reaction flask and cooled to 0 ° C. Then, a 2.5 mol / L solution of lithium aluminum hydride (6.48 mL, 16.2 mmol) in tetrahydrofuran was added, and the mixture was reacted at 0 ° C. for 2 hours. After confirming the completion of the reaction by TLC, the reaction was quenched by adding 5 mL of water, and then dried by adding a large amount of anhydrous sodium sulfate. The mixture was filtered, and the filter residue was thoroughly rinsed with dichloromethane. The filtrate was combined and concentrated by rotary evaporation. The residue was dried to obtain compound 1f (1.2 g, white solid). LC-MS: ESI [M+H] + =205.3; 1 H NMR(400MHz,DMSO-d6)δ11.87(s,1H),8.03(d,J=2.0Hz,1H),7.36(d,J=1.0Hz,1H),7 .34(dd,J=2.0,0.9Hz,1H),4.51(s,2H),2.52(d,J=1.8Hz,1H),1.15(t,J=7.4Hz,3H).

[0092] Step 6: Compound 1f (0.82 g, 4.0 mmol) was added to a 50 mL reaction flask, followed by the addition of 20 mL of dichloromethane and 1 mL of N,N-dimethylformamide. The mixture was cooled to 0° C., and then thionyl chloride (0.87 mL, 12 mmol) was added dropwise. The mixture was reacted at 0° C. for 1 hour. After the completion of the reaction was confirmed by TLC, the reaction liquid was concentrated by rotary evaporation. The crude product was purified by column chromatography to obtain 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).

[0093] 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. The mixture was stirred at 80° C. for 2 hours. After completion of the reaction was confirmed by TLC, the reaction mixture was concentrated under reduced pressure. The crude product was purified by column chromatography to give 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).

[0094] Embodiment 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 [ka]

[0095] Step 1: Compound int-1c (1.4 g, 4.5 mmol) was dissolved in 20 mL of MeOH. 5 mL of water was added, followed by lithium hydrate (570 mg, 13.5 mmol). The mixture was reacted at room temperature for 12 hours, and the progress was monitored by TLC. Upon completion of the reaction, the pH was adjusted to 6 by adding 2 M HCl. The mixture was extracted with EA (3 × 25 mL), and the combined organic phases were dried over anhydrous sodium sulfate and concentrated by rotary evaporation under vacuum to give product 2a (460 mg, pale yellow solid).

[0096] Step 2: Compound 2a (230 mg, 0.7 mmol), EDCl (191 mg, 1 mmol), HOBT (100 mg, 1 mmol), and 2 mL of DMF were added, followed by N-methylmorpholine (250 mg, 2.8 mmol) and d3-methylamine hydrochloride (49 mg, 0.7 mmol). The mixture was reacted at room temperature for 12 hours, and the progress was monitored by TLC. After complete consumption of the starting material, the mixture was diluted with water, extracted with EA, and the organic phase was washed five times with water, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation to give crude product 2b (150 mg, pale yellow oily liquid).

[0097] Step 3: The crude product 2b (220 mg, 0.7 mmol) was dissolved in 4 mL of MeOH, and then 1.1 mL of 4 M HCl was added to the dioxane solution. The mixture was allowed to react at room temperature for 12 hours, and the progress was monitored by TLC. Upon completion of the reaction, potassium carbonate was added and stirred for 30 minutes, and then filtered to remove the potassium carbonate, yielding crude compound 2c (270 mg, yellow-green solid).

[0098] Step 4: Compound 1f (40.6 mg, 0.2 mmol), 1 mL of DCM and DMF (15 mg, 0.02 mmol) were added, followed by thionyl chloride (70 mg, 0.6 mmol) at 0 °C. The mixture was allowed to react for 30 min, then continued at room temperature for another 30 min, and the progress was monitored by TLC. After complete consumption of the starting material, the mixture was concentrated by rotary evaporation under vacuum. Then, 2c (44.2 mg, 0.2 mmol) and DIPEA (180 mg, 1.4 mmol) were added. KI (10 mg, 0.06 mmol) was added, followed by 2 mL of acetonitrile. The mixture was heated to 80 °C, and the mixture was allowed to react for 2 h, and the progress was monitored by TLC. Upon completion, 2 mL of saturated sodium bicarbonate solution was added, extracted with EA (2 × 3 mL), and the organic phases were combined and purified by preparative TLC to give 2 (12 mg, pale yellow solid). LC-MS:ESI [M+H] + =407.51 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).

[0099] Embodiment 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 [ka]

[0100] Step 1: Compound 1f (0.4 g, 2.0 mmol) and dichloromethane (20 mL) were added to a 50 mL reaction flask and cooled to 0° C., and then 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 TLC confirmed the completion of the reaction, the mixture was filtered through diatomaceous earth, the filter cake was rinsed with dichloromethane, and the filtrate was concentrated by rotary evaporation and purified by column chromatography to give compound 3a (0.38 g, pale yellow solid). LC-MS: ESI [M+H] + =203.2; 1 H NMR (400MHz, DMSO-d6) δ12.17(s,1H),10.16(s,1H),8.92(d,J=1.8Hz,1H),7.85-7.83(m,2H),2.59(qd,J=7.4,1.3Hz,2H),1.21(t,J=7.4Hz,3H).

[0101] Step 2: Compound 3a (0.2 g, 1 mmol) was added to a reaction flask, which was then cooled to 0° C. with 15 mL of ether and 4 mL of methanol. 46 mg (1.1 mmol) of sodium borodeuteride was added, and the mixture was allowed to react at room temperature for 1 hour. After TLC confirmed the reaction was complete, the mixture was quenched with a small amount of water, concentrated by rotary evaporation, added a small amount of ice water to the slurry, filtered, and dried to give compound 3b (110 mg, white solid). LC-MS: ESI [M+H] + =206.2.

[0102] Step 3: Compound 3b (110 mg, 0.5 mmol) was added to a 25 mL reaction flask, followed by the addition of 10 mL of dichloromethane and 0.1 mL of N,N-dimethylformamide. The mixture was cooled to 0° C., and then thionyl chloride (88 mg, 0.75 mmol) was added dropwise. The mixture was reacted at 0° C. for 1 hour. After confirming the completion of the reaction by TLC, the reaction liquid was concentrated by rotary evaporation. The obtained crude product was purified by column chromatography to give compound 3c (103 mg, white solid). LC-MS: ESI [M+H] + =224.7.

[0103] 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 confirming the completion of the reaction by TLC, the reaction mixture was concentrated under reduced pressure, and the resulting crude product was purified by column chromatography to give compound 3 (60 mg, pale yellow solid). LC-MS: ESI [M+H] +=405.5;1H NMR(400MHz,DMSO)δ 11.85(s,1H),8.70(s,2 H),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).

[0104] Embodiment 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 [ka]

[0105] Step 1: Compound 4a (50 g, 314 mmol) and N-bromosuccinimide (67 g, 377 mmol) were added to a reaction flask, 300 mL of sulfuric acid was added, and the mixture was heated to 80 °C. The mixture was allowed to react overnight. After confirming the completion of the reaction by TLC, the reaction mixture was cooled to room temperature, and then the mixture was slowly diluted with ice water and extracted three times with ethyl acetate. The combined organic phases were washed successively with water and a saturated aqueous solution of sodium bicarbonate, dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 4b (65 g, pale yellow solid). LC-MS: ESI [M+H] + =238.9.

[0106] 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 the mixture was stirred and allowed to react overnight at room temperature. After confirming completion of the reaction by TLC, the reaction mixture was concentrated under reduced pressure, extracted three times with ethyl acetate and water, and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 4d (37 g, orange-red solid). LC-MS: ESI [M+H] + =336.1.

[0107] Step 3: Compound 4d (26 g, 78 mmol) was added to a mixture of 200 mL of anhydrous methanol and 5 mL of water, followed by the addition of ammonium chloride (35 g, 621 mmol). The mixture was cooled to 0°C, and zinc powder (43 g, 621 mmol) was slowly added. The mixture was heated to room temperature, and the mixture was allowed to react at room temperature for 1 hour. After TLC confirmed the completion of the reaction, the filtrate was filtered and concentrated under reduced pressure. Then, a dioxane solution (30 mL) containing 4 mol of hydrochloric acid was added, and the mixture was allowed to react at room temperature for 1 hour. After TLC confirmed the completion of the reaction, petroleum ether was added to precipitate a solid. After filtration and drying, compound 4e (16 g, off-white solid) was obtained. LC-MS: ESI [M+H] + =274.1.

[0108] 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 the mixture was allowed to react overnight at room temperature. After confirming the completion of the reaction by TLC, the reaction mixture was concentrated under reduced pressure, quenched with a saturated aqueous solution of sodium bicarbonate, extracted three times with ethyl acetate, and after completion of the organic phase, dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 4f (7.9 g, orange-red solid). LC-MS: ESI [M+H] +=272.1.

[0109] Step 5: Compound 4f (0.5 g, 1.8 mmol), tributylstannylmethanol (0.65 g, 2.0 mmol), and Xphos-Pd-G2 (73 mg, 0.09 mmol) were added, 15 mL of dioxane was added, the mixture was purged with nitrogen, and heated to 80 °C. The mixture was then reacted at 80 °C overnight. After confirming the completion of the reaction by TLC, water was added to the reaction liquid, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 4g (0.4 g, pale yellow solid). LC-MS: ESI [M+H] + =223.2.

[0110] Step 6: Compound 4g (0.4 g, 0.8 mmol) was added to 10 mL of aqueous hydrobromic acid and heated to 80 °C, and the mixture was allowed to react for 3 hours. After confirming the completion of the reaction by TLC, the reaction liquid was quenched with a saturated aqueous solution of sodium bicarbonate, extracted three times with dichloromethane, and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 4h (0.24 g, pale yellow solid). LC-MS: ESI [M+H] + =286.1.

[0111] 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 hours. After confirming the completion of the reaction by TLC, the reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give compound 4 (16 mg, pale 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).

[0112] Embodiment 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 [ka]

[0113] Step 1: Compound 2a (180 mg, 0.6 mmol), EDCl (144 mg, 0.75 mmol), HOBT (100 mg, 0.75 mmol), and 2 mL of DMF were added, followed by N-methylmorpholine (290 mg, 3.2 mmol) and cyclopropylamine (34 mg, 0.6 mmol). The mixture was reacted at room temperature for 12 hours, and the progress was monitored by TLC. After complete consumption of the starting material, the reaction mixture was diluted with water, extracted with EA, and the organic phase was washed five times with water, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation to give crude product 5a (180 mg, yellow oil).

[0114] Step 2: The crude product of 5a (180 mg, 0.6 mmol) was dissolved in 5 mL of MeOH, and then 0.9 mL of 4 M HCl was added to the dioxane solution. The mixture was allowed to react at room temperature for 12 hours, and the progress was monitored by TLC. After the reaction was complete, potassium carbonate was added and stirred for 30 minutes, and then filtered to remove the potassium carbonate, yielding crude compound 5b (180 mg, tan solid).

[0115] Step 3: Compound 3b (40.6 mg, 0.2 mmol), 1 mL of DCM and DMF (15 mg, 0.02 mmol) were added, followed by the addition of thionyl chloride (70 mg, 0.6 mmol) at 0 °C. The mixture was allowed to react at 0 °C for 30 minutes, then continued at room temperature for another 30 minutes, and the progress was monitored by TLC. After complete consumption of the starting material, the reaction mixture was concentrated by rotary evaporation under vacuum, and then 5b (44.2 mg, 0.2 mmol) and DIPEA (180 mg, 1.4 mmol) were added. KI (10 mg, 0.06 mmol) was added, followed by 2 mL of acetonitrile, and the mixture was heated to 80 °C. The mixture was allowed to react for 2 hours, and the progress was monitored by TLC. After the reaction was complete, 2 mL of saturated sodium bicarbonate solution was added, followed by extraction with EA (2 × 3 mL). The organic phases were combined and the reaction mixture was purified by TLC to give 5 (8 mg, pale yellow solid). LC-MS: ESI [M+H] + =431.2 1 H NMR(400MHz,DMSO)δ11.87(s,1H),8.69-8.65(m,2H),8.43(s,1H),8.04-7.94(m,2H),7.76(s,1H),7.66(s,1H),6.41(s,1H),3.76-3.66(m,1 H),3.17(s,2H),2.95-2.85(m,1H),2.77-2.68(m,3H),2.55(q,J=13.2Hz,2H),2.36-2.33(m,1H),1.19(t,J=13.2Hz,3H),0.75-0.61(m,4H).

[0116] Embodiment 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 [ka]

[0117] Step 1: 60% sodium hydride (16 g, 406 mmol) was added to DME (200 mL), purged with nitrogen three times, cooled to 0 °C, and compound 6a (76 g, 338 mmol) was added dropwise. The mixture was reacted at room temperature for 2 hours, and then deuterated methyl bromide (50 g, 439 mmol) was added dropwise. The reaction mixture was heated to 60 °C and reacted at 60 °C for 3 hours. After monitoring the progress of the reaction by HPLC, the reaction mixture was slowly poured into ice water to quench the reaction, extracted three times with ethyl acetate, and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The crude product, compound 6c (83 g, colorless liquid), was obtained.

[0118] Step 2: 60% sodium hydride (13 g, 322 mmol) was added to the reaction flask, followed by 200 mL of tetrahydrofuran. The mixture was then purged with nitrogen three times, cooled to 0 ° C., and compound 6c (83 g, 322 mmol) was slowly added dropwise. The mixture was stirred and reacted at 0 ° C. for 10 minutes. The reaction mixture was then heated to room temperature and stirred. The mixture was allowed to react at room temperature for another 10 minutes. The reaction mixture was further heated to 40 ° C. and finally cooled to -78 ° C. for 5 minutes after the reaction. A solution of compound 1b (48 g, 215 mmol) dissolved in 200 mL of tetrahydrofuran was slowly added dropwise, and the mixture was stirred and reacted at -78 ° C. for 1 hour. After confirming the completion of the reaction by TLC, the reaction mixture was slowly quenched by adding ice-cold saturated aqueous ammonium chloride solution to it, extracted three times with ethyl acetate, and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 6d (41 g, yellow-green liquid). LC-MS: ESI [M+H] + =328.4.

[0119] Step 3: Compound 6d (1.0 g, 3.0 mmol) was added to 10 mL of absolute ethyl alcohol, followed by the addition of Pd / C (0.1 g, 10%). The mixture was purged with hydrogen three times, stirred, and allowed to react at room temperature overnight. After confirming the completion of the reaction by LC-MS, the reaction liquid was filtered, the filter residue was thoroughly rinsed with ethanol, and the combined filtrate was concentrated by rotary evaporation. A solution of 4 mol / L hydrochloric acid (12 mL) in 1,4-dioxane was added and stirred at room temperature for 30 minutes. Diethyl ether was added to precipitate a large amount of solid. The solid was filtered and dried to give compound 6e (0.51 g, white solid). LC-MS: ESI [M+H] + =254.3.

[0120] Step 4: Compound 6e (0.51 g, 2.0 mmol) was added to a 50 mL reaction flask, followed by 15 mL of 1,4-dioxane. DDQ (0.50 g, 2.2 mmol) was then added and refluxing began. The mixture was allowed to react overnight. After LC-MS confirmed the completion of the reaction, the reaction liquid was concentrated by rotary evaporation. A saturated aqueous solution of sodium bicarbonate was added and stirred for 1 hour. The liquid was filtered, and the residue was washed with water and then rinsed with a small amount of ether. The residue was dried to give compound 6f (0.3 g, yellow solid). LC-MS: ESI [M+H] + =252.3.

[0121] Step 5: Compound 6f (0.3 g, 1.2 mmol) and 15 mL of tetrahydrofuran were added to a reaction flask and cooled to 0° C. Then, a 2.5 mol / L solution of lithium aluminum hydride in tetrahydrofuran (1.44 mL, 3.6 mmol) was added. The mixture was reacted at 0° C. for 2 hours. After the completion of the reaction was confirmed by TLC, 1 mL of water was added to quench the reaction. A large amount of anhydrous sodium sulfate was added for drying, followed by filtration. The filter residue was rinsed with a large amount of dichloromethane. After combining the filtrates, the residue was concentrated by rotary evaporation and then dried to obtain compound 6g (0.2 g, yellow solid). LC-MS: ESI [M+H] + =210.3.

[0122] Step 6: Compound 6g (0.2 g, 0.96 mmol) was added to a 25 mL reaction flask, followed by the addition of 10 mL of dichloromethane and 0.1 mL of N,N-dimethylformamide. The mixture was cooled to 0°C, and then thionyl chloride (0.35 g, 2.9 mmol) was added dropwise. The mixture was reacted at 0°C for 1 hour. After confirming the completion of the reaction by TLC, the reaction liquid was concentrated by rotary evaporation. The obtained crude product was purified by column chromatography to obtain compound 6h (0.2 g, gray solid). LC-MS: ESI [M+H] + =228.7.

[0123] Step 7: Compound 6h (0.2 g, 0.88 mmol), int-1 (0.25 g, 0.88 mmol), N,N-diisopropylethylamine (0.45 g, 3.52 mmol), and potassium iodide (15 mg, 0.09 mmol) were added to 10 mL of anhydrous acetonitrile. The mixture was stirred at 85° C. for 2 hours. After completion of the reaction was confirmed by TLC, the reaction mixture was concentrated under reduced pressure. The resulting 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)δ 10.97(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=8.2,2.3 Hz,1H),7.66(s,1H),6.23(s,1H),3.78(s,2H),3.27(d,J=3.1Hz,2H),3.04(d,J=5.1Hz,3H),2.79(t,J=5.6Hz,2H),2.60(d,J=1.4Hz,2H).

[0124] Embodiment 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 [ka]

[0125] Step 1: Compound 7a (0.5 g, 2.2 mmol), int-1b (2.4 g, 5.5 mmol), Pd(dppf)Cl (0.16 g, 0.22 mmol), and potassium carbonate (0.7 g, 5.5 mmol) were added to a mixture of 7 mL of dioxane, 3 mL of anhydrous ethyl alcohol, and 4 mL of water. The mixture was then purged with nitrogen three times, and reacted under nitrogen protection at 90 °C for 2 h. After confirming completion of the reaction by TLC, the reaction mixture was cooled to room temperature, and 30 mL of dichloromethane and 20 mL of water were added. The layers were separated using a separatory funnel. The aqueous phase was extracted twice with dichloromethane and then washed sequentially with water and saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 7b (0.7 g, white solid).

[0126] 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 and allowed to react at room temperature overnight. After completion of the reaction was confirmed by TLC, the reaction mixture was concentrated under reduced pressure to give dried compound 7c (0.5 g, white solid).

[0127] Step 3: Compound 7c (0.5 g, 1.7 mmol) was added to 10 mL of anhydrous methanol followed by 10 mL of a solution of hydrochloric acid and dioxane (4 mol / L), and the mixture was stirred at room temperature for 0.5 to 1 h. After completion of the reaction was confirmed by TLC, the reaction mixture was concentrated under reduced pressure to give dried compound 7d (0.3 g, white solid).

[0128] 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 the mixture was stirred at 80 °C for 2 hours. After confirming the completion of the reaction by TLC, the reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give 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).

[0129] Embodiment 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 [ka]

[0130] The compound 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-2-methyl-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was obtained via a similar procedure as described in embodiment 2. LC-MS: ESI [M+H] + =421.2. 1H 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).

[0131] Embodiment 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 [ka]

[0132] 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, and then purged with nitrogen three times. The mixture was stirred at room temperature for 48 hours under nitrogen protection. After completion of the reaction was confirmed by TLC, the reaction mixture was concentrated to dryness to obtain the crude product, which was purified by column chromatography to obtain compound 9a (2.2 g, white solid).

[0133] Step 2: Compound 9a (1 g, 4.3 mmol), int-1b (1.59 g, 5.1 mmol), Pd(dppf)Cl (0.31 g, 0.43 mmol), and potassium carbonate (1.5 g, 10.8 mmol) were added to a mixture of 7 mL of dioxane, 3 mL of anhydrous ethyl alcohol, and 4 mL of water. The mixture was then purged with nitrogen three times and reacted at 90 °C under nitrogen protection for 2 h. After confirming the completion of the reaction by TLC, the reaction mixture was cooled to room temperature, and 30 mL of dichloromethane and 20 mL of water were added. The layers were separated using a separatory funnel. The aqueous phase was extracted twice with dichloromethane and then washed sequentially with water and saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 9b (1.2 g, white solid).

[0134] Step 3: Compound 9b (1 g, 2.9 mmol), aqueous methylamine solution (5 g, 161.3 mmol), and anhydrous methanol (20 mL) were added to a 100 mL reaction flask, and the mixture was stirred and reacted at room temperature overnight. After completion of the reaction was confirmed by TLC, the reaction mixture was concentrated under reduced pressure to give dried compound 9c (0.8 g, white solid).

[0135] Step 4: Compound 9c (0.5 g, 1.4 mmol) was added to 10 mL of anhydrous methanol followed by 10 mL of a 4 mol / L solution of hydrochloric acid and dioxane, and the mixture was stirred at room temperature for 0.5 to 1 h. After completion of the reaction was confirmed by TLC, the reaction mixture was concentrated under reduced pressure to give compound 9d (0.3 g, a white solid).

[0136] 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 the mixture was stirred at 80 °C for 2 hours. After confirming the completion of the reaction by TLC, the reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give 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).

[0137] Embodiment 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 [ka]

[0138] The compound 1'-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-2-fluoro-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was obtained via a similar procedure as described in embodiment 2. LC-MS: ESI [M+H] + =425.2. 1H NMR(400MHz,Chloroform-d)δ11.15(s,1H),8.47(d,J=1.8Hz,1H),7.99(dd,J=7.7,1.7Hz,1H),7.83-7.73(m,2H),7.63(s,1H) ,7.57(s,1H),6.13(t,J=2.6Hz,1H),3.72(s,2H),3.21(d,J=3.3Hz,2H),2.75-2.62(m,4H),2.52(s,2H),1.24(t,J=7.4Hz,3H).

[0139] Embodiment 11: 1'-((7-ethyl-d5)-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide [ka]

[0140] The compound 1'-((7-(ethyl-d5)-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)-N-(methyl-d3)-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide was obtained via a similar procedure as described in embodiment 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).

[0141] Embodiment 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. [ka]

[0142] The compound 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 was obtained via a similar procedure as described in embodiment 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).

[0143] Embodiment 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 [ka]

[0144] Step 1: Compound 13a (10.0 g, 39.5 mmol) was added to a reaction flask and dissolved in 100 mL of DMF under nitrogen protection. Then, t-BuOK (6.6 g, 59.2 mmol) was added to the system in batches at -10 °C and stirred continuously for 1 h at -10 °C. After the addition, the mixture was slowly added dropwise to deuterated methyl bromide (5.0 g, 43.4 mmol), and the mixture was allowed to warm to room temperature and stirred for 3 h. 300 mL of water was added to the system, and the organic phase was extracted with EA (500 mL x 3). The organic phases were combined and washed with saturated sodium chloride solution. The liquid was separated, dried over anhydrous Na2SO4, and concentrated in vacuo to give yellow oily liquid 13b (11.0 g, 97%). LC-MS: ESI [M+H] + =287.2.

[0145] Step 2: Compound 13b (11.0 g, 38.4 mmol), 120 mL of a mixed solvent of EA / HO (3 / 1), and 4 M HCl-ethyl acetate solution (28.8 mL, 115.2 mmol) were added. The mixture was stirred and reacted overnight at room temperature, concentrated in vacuo to remove the solvent, and EA was added to form a slurry. The resulting white solid was the hydrochloride salt of 13c (2.5 g, 41%). LC-MS: ESI [M+H] + =123.1.

[0146] Step 3: The hydrochloride salt of 13c (2.5 g, 15.8 mmol) was added to a reaction flask, and 30 mL of DMF was added to dissolve the hydrochloride salt of 13c. DIEA (8.1 g, 63.0 mmol) was added to the reaction flask and stirred at room temperature, followed by 4b (4.1 g, 17.3 mmol). The mixture was stirred at room temperature for 5 hours under nitrogen protection. After the reaction was completed, 90 mL of pure water was added, and the organic phase was extracted with EA (150 mL x 2). The combined organic phase was washed with saturated sodium chloride solution, the liquid was separated, and concentrated under vacuum. The organic phase was purified by column chromatography with PE and DCM (0%-20%) to give yellow oil 13d (4.8 g, 90%). LC-MS: ESI [M+H] + =340.0.

[0147] Step 4: Compound 13d (4.5 g, 13.2 mmol) was dissolved in 80 mL of glacial acetic acid, and the system was stirred at room temperature when reduced Fe powder (4.4 g, 79.4 mmol) was slowly added. The mixture was stirred at 70 °C for 1 h and filtered before cooling. The filter cake was washed with a mixed solvent of DCM and MeOH, and the filtrate was concentrated in vacuo and purified by column chromatography using PE and EA (0%-25%) to give a white solid 13e (2.1 g, 57%). LC-MS: ESI [M+H] + =278.0.

[0148] Step 5: Compound 13e (2.1 g, 7.6 mmol) was added to a reaction flask, 30 mL of DCM was added, and the mixture was stirred. Then, DDQ (2.1 g, 9.1 mmol) was slowly added. The mixture was stirred and reacted at room temperature overnight. After the reaction was completed, a saturated aqueous solution of NaHCO3 was added to the system, the liquid was separated, and the aqueous phase was extracted with a mixed solvent of DCM and MeOH (5:1) (30 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated. The dried filtrate was purified by column chromatography using DCM and MeOH (0%-3%) to obtain a white solid 13f (2.0 g, 96%). LC-MS: ESI [M+H] + =276.0.

[0149] Step 6: Compound 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 placed in a reaction flask, and 30 mL of dioxane was added. Nitrogen was purged 3 to 5 times, and the mixture was stirred at 90 °C under nitrogen protection for 4 h. After the reaction was complete, the mixture was concentrated under vacuum until the solvent was removed. The concentrate was purified by column chromatography using DCM and MeOH (0%-5%) to give a white solid 13g (946.0 mg, 76%). 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=7.7 Hz,1H),5.46(t,J=5.8Hz,1H),4.68(d,J=5.6Hz,2H).

[0150] Step 7: Compound 13g (650 mg, 2.9 mmol) was added to a reaction flask, and 20 mL of DCM was added to dissolve 13g. Dess-Martin oxidant (1.5 g, 3.4 mmol) was slowly added at 0°C, and the mixture was stirred at 0°C for 0.5 h. After the reaction was completed, a saturated aqueous solution of NaHCO3 was added to the system and stirred for 10 minutes. The liquid was separated, and the organic solvent was concentrated to dryness under vacuum. The concentrate was purified by column chromatography using DCM and MeOH (0%-5%) to give a white solid 13h (610 mg, 95%). LC-MS: ESI [M+H] + =226.1; 1 H NMR (400MHz, CDCl3) δ 10.37 (s, 1H), 9.85 (s, 1H), 7.72-7.63 (m, 2H).

[0151] Step 8: Compound int-1a (99.2 mg, 0.34 mmol) was added to a reaction flask, and 5 mL of DCM was added first, followed by the addition of TEA to release the hydroxyl groups, followed by the dropwise addition of glacial acetic acid until the pH reached approximately 5. Finally, 13h (70.0 mg, 0.31 mmol) was added, and the mixture was stirred at room temperature for 2 h. Sodium triacetoxyborohydride (263.5 mg, 1.24 mmol) was added to the system and stirred for 2 h, followed by the addition of a saturated aqueous solution of NH4Cl and stirring. The liquid was separated, and the aqueous phase was extracted with a mixed solvent of DCM and MeOH (5:1) (30 mL x 3). The combined organic phases were dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness. The residue was treated by pre-HPLC and lyophilized to give compound 13 (16 mg, 12%) as a white solid. LC-MS: ESI [M+H] + =427.2; 1H 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).

[0152] Embodiment 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 [ka]

[0153] Compound 9d (150.5 mg, 0.49 mmol) was added to a reaction flask, and 5 mL of DCM was added first, followed by the addition of TEA to release the HCl residue, followed by the dropwise addition of glacial acetic acid until the pH reached approximately 5. Finally, 13h (100.0 mg, 0.44 mmol) was added, and the mixture was stirred at room temperature for 2 h. Sodium triacetoxyborohydride (376.4 mg, 1.78 mmol) was added to the system and stirred for 2 h. A saturated aqueous solution of NH₄Cl was then added and stirred. The liquid was separated, and the aqueous phase was extracted with a 5:1 mixture of DCM and MeOH (30 mL x 3). The combined organic phases were dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated to dryness. The residue was treated by pre-HPLC and lyophilized to give compound 14 (60 mg, 30%) as a white solid. LC-MS: ESI [M+H] + =445.2; 1H 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).

[0154] Embodiment 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 [ka]

[0155] Step 1: Compound 1e (2.0 g, 8.1 mmol) was added to a 150 mL reaction flask, followed by 60 mL of tetrahydrofuran. The mixture was cooled to 0 °C, and then a 2.5 mol / L solution of lithium aluminum deuteride was dissolved in tetrahydrofuran (6.48 mL, 16.2 mmol) and reacted at 0 °C for 2 h. After confirming the completion of the reaction by TLC, 5 mL of water was added to quench the reaction, followed by adding a large amount of anhydrous sodium sulfate to dry the mixture. The mixture was filtered, the filter cake was washed with a large amount of dichloromethane, and the combined filtrate was concentrated by rotary evaporation. The residue was dried to give compound 15a (0.8 g, white solid). LC-MS: ESI [M+H] + =207.3.

[0156] Step 2: Compound 15a (0.82 g, 4.0 mmol) was added to a 50 mL reaction flask, followed by 20 mL of dichloromethane and 1 mL of N,N-dimethylformamide. The mixture was cooled to 0 °C, and thionyl chloride (0.87 mL, 12 mmol) was added dropwise. The reaction was allowed to proceed at 0 °C for 1 hour. After confirming the completion of the reaction by TLC, the reaction mixture was concentrated by rotary evaporation, and the resulting crude product was purified by column chromatography to obtain compound 15b (0.52 g, gray solid). LC-MS: ESI [M+H] + =225.7.

[0157] 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 hours. After confirming the completion of the reaction by TLC, the reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give compound 15 (25 mg, white solid). LC-MS: ESI [M+H] + =406.4.

[0158] Embodiment 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 [ka]

[0159] Step 1: Compound 16a (500.0 g, 1.99 mol) and 2.5 L of methanol were added and stirred under nitrogen protection. The mixture was cooled to 0°C-5°C. Sodium methoxide (118.0 g, 2.19 mol) and 1.0 L of methanol were added and dissolved by stirring. The mixture was then added dropwise to the reaction mixture while controlling the temperature at 0°C-5°C. After the addition was complete, the mixture was allowed to warm to room temperature and stirred for 1 h. After confirming the completion of the reaction by TLC, 2.0 L of water was added to the reaction mixture. The reaction mixture was concentrated under reduced pressure at 40°C-50°C until no liquid was generated. The aqueous layer was extracted twice with ethyl acetate (4.0 L and 1.0 L, respectively). The combined organic phases were concentrated under reduced pressure at 40°C-50°C until the weight remained constant, yielding compound 16b (480 g, white solid).

[0160] Step 2: Compound 16b (475 g, 1.93 mol), 2.85 L of DMF, and 2.85 L of DMF-DMA were added, and the mixture was heated to 100°C and stirred for 2 hours. After confirming the completion of the reaction by TLC, the reaction mixture was concentrated at 70°C-80°C until all the liquid was removed. 10.0 L of water was added, and the mixture was stirred at 20°C-30°C for 2 hours. The mixture was then filtered. The filter cake was dried under reduced pressure at 60°C-70°C until its weight remained constant, yielding compound 16c (612 g, red solid).

[0161] Step 3: Compound 16c (500 g, 1.91 mol) and 2.56 L of tetrahydrofuran were added. Sodium periodate (805.0 g, 3.72 mol) was dissolved in 2.56 L of water. The sodium periodate solution was added dropwise to the reaction mixture at 25°C to 30°C and stirred for 2 to 4 h at 25°C to 30°C. After confirming the completion of the reaction by TLC, 2.56 L of water and 4.0 L of ethyl acetate were added, and the mixture was stirred and the layers were separated. The aqueous layer was washed twice with 2.0 L of ethyl acetate. The organic layers were combined and washed successively with 4.0 L of saturated sodium thiosulfate solution and 4.0 L of saturated sodium chloride solution. The liquid was separated, and the organic layer was concentrated under reduced pressure at 40°C to 50°C until its weight remained constant, to obtain compound 16d (500 g, oil).

[0162] Step 4: Compound 16d (500.0 g, 1.69 mol) and 3,3-dideoxypropionic acid ethyl ester (1,457.0 g, 7.61 mol) were added to 7.5 L of anhydrous ethyl alcohol, and stannous chloride (1,815.0 g, 9.57 mol) was added. After the addition, the temperature was increased to reflux for 1 to 2 h. After confirming the completion of the reaction by TLC, the reaction mixture was concentrated under reduced pressure at 40 to 50 °C until the weight remained constant. 15.0 L of ethyl acetate was added, and the pH was adjusted to 7 to 8 with saturated sodium bicarbonate solution. The organic layer was filtered and concentrated under reduced pressure at 40 to 50 °C until the weight remained constant. The mixture was purified by column chromatography to obtain compound 16e (230.0 g, white solid).

[0163] Step 5: Compound 16e (80.0 g, 257.1 mmol) and 960 mL of tetrahydrofuran were added, and the mixture was cooled to 0°C in an ice-water bath. Diisopropylaluminum hydride (1 M, 771.3 mL) was added dropwise, and the reaction temperature was raised by 20°C to 25°C after the addition. After confirming the completion of the reaction by LC-MS, 800 mL of saturated ammonium chloride solution was added to the reaction system, and the filtrate was filtered and concentrated by rotary evaporation. 80 mL of dichloromethane was added to the pulp, and the filtrate was filtered and concentrated under reduced pressure at 40°C to 50°C until the weight remained constant, to give compound 16f (43.0 g, yellow solid).

[0164] Step 6: Compound 16f (45.0 g, 167.2 mmol), DIPEA (64.8 g, 501.6 mmol), sodium iodide (5.0 g, 33.4 mmol), and 450 mL of tetrahydrofuran were added. MOMCl (26.9 g, 334.4 mmol) was added dropwise while stirring. The mixture was then heated to 45-50°C, and the reaction was stirred for 3 h. After TLC confirmed the reaction was complete, the mixture was cooled to 20-25°C, 500 mL of water was added, and the liquid was separated. The aqueous layer was extracted with 300 mL of ethyl acetate, and the organic layers were combined. The organic layers were concentrated under reduced pressure at 40-50°C until dry and purified by column chromatography to give compound 16g (34.8 g, yellow solid).

[0165] Step 7: Compound 16g (39.8 g, 127.1 mmol), triethylamine (15.5 g, 152.5 mmol), and Pd(dppf)Cl (9.22 g, 12.7 mmol) were added, followed by 800 mL of methanol. The atmosphere was purged with nitrogen three times, followed by carbon monoxide three times. The mixture was heated to 60 °C and reacted for 12 h. After confirming the completion of the reaction by LC-MS, the reaction mixture was concentrated under reduced pressure at 40 °C to 50 °C to remove the solvent. The residue was purified by column chromatography to give compound 16h (32.0 g, white solid).

[0166] Step 8: Compound 16h (32.0 g, 109.5 mmol) and 384 mL of tetrahydrofuran were added, and the mixture was cooled to 0°C in an ice-water bath. Diisopropylaluminum (1 M, 328.5 mmol) was added dropwise, and the reaction temperature was then returned to 20°C-25°C after the addition. After confirming the completion of the reaction by LC-MS, 320 mL of saturated ammonium chloride solution and 320 mL of ethyl acetate were added to the reaction system, and the filtrate was filtered and concentrated under reduced pressure at 40°C-50°C to obtain compound 16i (28.6 g, yellow solid).

[0167] Step 9: Compound 16i (28.6 g, 108.2 mmol) and 286 mL of dichloromethane were added and stirred to completely dissolve. The mixture was cooled to 0°C in an ice-water bath, and then Dess-Martin periodinane (55.0 g, 129.8 mmol) was added in batches. After the addition, the temperature was returned to 20°C-25°C, and the reaction was allowed to proceed for 2 hours. After TLC confirmed the completion of the reaction, 300 mL of saturated sodium bicarbonate solution and saturated sodium thiosulfate (1:1) were added, and the mixture was washed twice. The aqueous layer was extracted with 300 mL of dichloromethane, and the organic layers were combined and concentrated under reduced pressure at 40°C-50°C. The mixture was purified by column chromatography to give compound 16j (23.4 g, yellow solid).

[0168] Step 10: Compound 16j (23.4 g, 89.1 mmol) and 234 mL of tetrahydrofuran were added, and the mixture was cooled to -40°C in a dry ice bath. Deuterated methylmagnesium iodide (1 M, 106.9 mmol) was added dropwise, and the mixture was allowed to warm to room temperature and stirred for 1 h. After confirming the completion of the reaction by TLC, the reaction was quenched by adding 234 mL of saturated ammonium chloride solution and 234 mL of water, and the mixture was stirred and separated. The organic layers were combined and concentrated under reduced pressure at 40°C to 50°C to give compound 16k (25.6 g, oil).

[0169] Step 11: Compound 16k (24.7 g, 88.1 mmol) and 247 mL of dichloromethane were added and stirred to completely dissolve. The mixture was cooled to 0°C in an ice-water bath, and then Dess-Martin periodinane (44.8 g, 105.7 mmol) was added in batches. After the addition, the temperature was returned to 20°C-25°C, and the mixture was allowed to react for 2 hours. After confirming the completion of the reaction by TLC, 300 mL of saturated sodium bicarbonate solution and saturated sodium thiosulfate (1:1) were added, and the mixture was washed twice. The aqueous layer was extracted with 300 mL of dichloromethane, and the organic layers were combined and concentrated under reduced pressure at 40°C-50°C until dry. The mixture was purified by column chromatography to give compound 16l (23.2 g, yellow solid).

[0170] Step 12: Compound 16l (5.0 g, 17.9 mmol), p-toluenesulfonylhydrazide (3.3 g, 17.9 mmol), and 100 mL of methanol were added, and the mixture was reacted at 20° C. to 25° C. for 16 hours. After confirming the completion of the reaction by TLC, the reaction liquid was filtered, and the filter cake was dried under reduced pressure at 40° C. to 50° C. until its weight remained unchanged to obtain compound 16m (7.20 g, white solid).

[0171] Step 13: Compound 16m (4.50 g, 10.0 mmol) and 45 mL of dichloromethane were added, the mixture was cooled to 0 °C in an ice-water bath, and diisopropylaluminum hydride (1 M, 20 mL) was added dropwise. The mixture was heated to 20 °C-25 °C for the reaction. After the completion of the reaction was confirmed by LC-MS, 20 mL of saturated ammonium chloride solution was added to the reaction system, and the filtrate was filtered and concentrated by rotary evaporation. The resulting product was purified by column chromatography to obtain compound 16n (540 mg, oil).

[0172] Step 14: Compound 16n (400.0 mg, 1.51 mmol), 4 mL of dioxane, and 4 mL of 48% HBr / HO were added, and the mixture was heated to 80° C. and reacted for 2 hours. After the completion of the reaction was confirmed by LC-MS, the pH was adjusted to 7-8 using saturated sodium carbonate solution, and the filter cake was filtered and purified by column chromatography to obtain compound 16o (200 mg, white solid).

[0173] Step 15: Compound 16o (100.0 mg, 0.48 mmol), 3 mL of toluene, and DMF (3.5 mg, 0.048 mmol) were added, and the mixture was cooled to 0°C in an ice-water bath. Thionyl chloride (69.0 mg, 0.48 mmol) was added dropwise, and then the mixture was heated to 20°C-25°C to react. After completion of the reaction was confirmed by TLC-MS, the mixture was concentrated under reduced pressure to remove the solvent, yielding compound 16p (108 mg, yellow solid).

[0174] Step 16: Compound 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. The mixture was heated to 80 °C and reacted for 2 h. After completion of the reaction was confirmed by LC-MS, the mixture was cooled to room temperature and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 16 (100 mg, white solid). LC-MS: ESI [M+H] +=407.2;1H NMR(400MHz,DMSO-d6)δ 11.89(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=3.6 Hz,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).

[0175] Embodiment 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-carboxamide [ka]

[0176] 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 mixture of 7 mL of dioxane, 3 mL of anhydrous ethyl alcohol, and 4 mL of water. The mixture was then purged with nitrogen three times and reacted at 90 °C under nitrogen protection for 2 h. After confirming completion of the reaction by TLC, the reaction mixture was cooled to room temperature, and 30 mL of dichloromethane and 20 mL of water were added. The layers were separated using a separatory funnel. The aqueous phase was extracted twice with dichloromethane, and the combined organic phases were washed sequentially with water and saturated brine. The reaction mixture was dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 17b (0.7 g, white solid).

[0177] Step 2: Compound 17b (0.5 g, 1.8 mmol) was added to 10 mL of anhydrous methanol, followed by 10 mL of a 4 mol / L solution of hydrochloric acid and dioxane, and the mixture was stirred at room temperature for 0.5 to 1 h. After confirming the completion of the reaction by TLC, the reaction mixture was concentrated under reduced pressure to give compound 17c (0.2 g, a white solid).

[0178] 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 hours. After confirming the completion of the reaction by TLC, the reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give 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).

[0179] Embodiment 18: 3-(ethyl-2,2,2-d3)-7-((5-fluoro-3',6'-dihydro-[2,4-bipyridin]-1'(2'H)-yl)methyl)-1,5-naphthyridin-2(1H)-one [ka]

[0180] The compound 3-(ethyl-2,2,2-d3)-7-((5-fluoro-3',6'-dihydro-[2,4-bipyridin]-1'(2'H)-yl)methyl)-1,5-naphthyridin-2(1H)-one was obtained via a procedure similar to that described in 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).

[0181] Embodiment 19: 3-(ethyl-d5)-8-fluoro-7-((5-fluoro-3',6'-dihydro-[2,4-bipyridin]-1'(2'H)-yl)methyl)quinoxalin-2(1H)-one [ka]

[0182] Compound 18c (122.6 mg, 0.49 mmol) was added to the reaction flask, and 5 mL of DCM was added. Then, TEA was added to release the eluate, followed by dropwise addition of glacial acetic acid until the pH reached approximately 5. Finally, 13h (100.0 mg, 0.44 mmol) was added, and the mixture was stirred at room temperature for 2 h. Sodium triacetoxyborohydride (376.4 mg, 1.78 mmol) was added to the system and stirred for 2 h. A saturated aqueous solution of NH4Cl was added and stirred. The liquid was separated, and the aqueous phase was extracted with a mixed solvent of DCM and MeOH (5:1) (30 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness. The residue was treated by preparative HPLC and lyophilized to give compound 19 (32 mg, 19%) as a white solid. LC-MS: ESI [M+H] + =388.2; 1H 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).

[0183] Embodiment 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 [ka]

[0184] Compound 17c (126.1 mg, 0.49 mmol) was added to the reaction flask, and 5 mL of DCM was added first, followed by the addition of TEA to release the HCl, followed by the dropwise addition of glacial acetic acid until the pH reached approximately 5. Finally, 13h (100.0 mg, 0.44 mmol) was added, and the mixture was 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. A saturated aqueous solution of NH4Cl was added and stirred. The liquid was separated, and the aqueous phase was extracted with a mixed solvent of DCM and MeOH (5:1) (30 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness. The residue was treated by preparative HPLC and lyophilized to give compound 20 (14 mg, 8%) as a white solid. LC-MS: ESI [M+H] + =395.20; 1H 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).

[0185] Embodiment 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 [ka]

[0186] Step 1: Compound 21a (6.0 g, 49.2 mmol) was added to 50 mL of tetrahydrofuran, and the mixture was cooled to -78 °C. LDA (2 M, 29.5 mL, 59.0 mmol) was slowly added dropwise while maintaining the temperature at -60 °C to -78 °C. After the addition was complete, the mixture was stirred for 15 minutes, and then 40 mL of a tetrahydrofuran solution of 21b (19.3 g, 54.1 mmol) was added dropwise. The reaction mixture was allowed to warm to room temperature and stirred for 2 hours. After confirming the completion of the reaction by TLC, 60 mL of saturated aqueous ammonium chloride and saturated ammonium chloride were added, stirred, and the mixture was separated. The aqueous layer was then extracted with 100 mL of ethyl acetate, and the mixture was separated again. The organic layers were combined and washed with 100 mL of saturated sodium chloride solution. After the phase separation was complete, the organic layer was concentrated under reduced pressure and the solvent was removed to give compound 21c (20.0 g, oil).

[0187] Step 2: Compound 21c (19.3 g, 28.7 mmol), potassium carbonate (7.93 g, 57.4 mmol), and 21d (8.32 g, 31.6 mmol) were added to a solution of 100 mL of 1,4-dioxane / water (10:1). The atmosphere was purged with nitrogen three times, and the mixture was then heated to 80 °C and stirred for 2 h. After completion of the reaction was confirmed by LC-MS, the mixture was cooled to room temperature. 100 mL of water and 100 mL of ethyl acetate were added, and the layers were separated. The aqueous layer was extracted three times with 100 mL of ethyl acetate. The organic layers were combined and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 21e (4.2 g, yellow solid).

[0188] Step 3: Compound 21e (4.2 g, 13.0 mmol) was added to 21 mL of anhydrous methanol, followed by the addition of 40% aqueous methylamine (5.1 g, 65.2 mmol) and stirring at room temperature for 1 hour. After completion of the reaction was confirmed by LC-MS, the reaction mixture was concentrated to dryness. 40 mL of 1,4-dioxane was added, followed by the dropwise addition of 4 M hydrochloric acid and dioxane solution (16.3 mL, 65.2 mmol). The mixture was stirred at room temperature for 2 hours. After completion of the reaction was confirmed by LC-MS, the filtrate was filtered and concentrated by rotary evaporation to give compound 21f (3.4 g, off-white solid).

[0189] 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. The mixture was heated to 80° C. and stirred for 2 hours. After the completion of the reaction was confirmed by LC-MS, 200 mL of saturated sodium bicarbonate solution was added and stirred. The filtrate was filtered, concentrated by rotary evaporation, and purified by column chromatography to give compound 21 (2.4 g, white solid). LC-MS: ESI [M+H] + =408.2; 1H NMR(400MHz,DMSO-d6)δ 11.90(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).

[0190] Embodiment 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 [ka]

[0191] Step 1: Compound 21c (20 g, 60 mmol), pinacol diboronate (15.2 g, 72 mmol), potassium acetate (11.8 g, 120 mmol), and PdCl(dppf) (2.1 g, 3 mmol) were sequentially added to a reaction flask and reacted overnight at 80° C. under N protection. After the reaction was completed, the reaction liquid was filtered and washed with EA. The filtrate was concentrated, and the residue was purified by column chromatography to give compound 22a (15 g, oil).

[0192] Step 2: Compound 22a (15 g, 22.3 mmol), potassium carbonate (7.93 g, 57.4 mmol), and 9a (8.32 g, 31.6 mmol) were added to a solution of 100 mL of 1,4-dioxane / water (10:1). The atmosphere was purged with nitrogen three times, and the mixture was then heated to 80 °C and stirred for 2 h. After completion of the reaction was confirmed by LC-MS, the mixture was cooled to room temperature. 100 mL of water and 100 mL of ethyl acetate were added, and the layers were separated. 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 concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 22b (12 g, yellow solid).

[0193] Step 3: Compound 22b (4.2 g, 13.0 mmol) was added to 21 mL of anhydrous methanol, followed by the addition of 40% aqueous methylamine (5.1 g, 65.2 mmol) and stirring at room temperature for 1 h. After confirming completion of the reaction by LC-MS, the reaction mixture was concentrated to dryness. 40 mL of 1,4-dioxane was added, followed by the dropwise addition of 4 M hydrochloric acid and dioxane solution (16.3 mL, 65.2 mmol) and stirring at room temperature for 2 h. After confirming completion of the reaction by LC-MS, the filtrate was filtered and concentrated by rotary evaporation to give compound 22c (3.4 g, off-white solid).

[0194] 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. The mixture was heated to 80 °C and stirred for 2 h. After the completion of the reaction was confirmed by LC-MS, 200 mL of saturated sodium bicarbonate solution was added and stirred. The filtrate was filtered, concentrated by rotary evaporation, and purified by column chromatography to give compound 22 (2.4 g, white solid). LC-MS: ESI [M+H] + =426.5; 1H NMR(400MHz,DMSO)δ 11.84(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).

[0195] Embodiment 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 [ka]

[0196] Step 1: Compound 22a (13 g, 21.2 mmol), potassium carbonate (7.8 g, 55 mmol), and 100 mL of 7a (8.1 g, 28.6 mmol) were added to a solution of 100 mL of 1,4-dioxane / water (10:1). The atmosphere was purged with nitrogen three times, and the mixture was then heated to 80 °C and stirred for 2 h. After completion of the reaction was confirmed by LC-MS, the mixture was cooled to room temperature. 100 mL of water and 100 mL of ethyl acetate were added, and the layers were separated. The aqueous layer was extracted three times with 100 mL of ethyl acetate. The organic layers were combined and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated by rotary evaporation. The resulting crude product was purified by column chromatography to give compound 23a (10.6 g, yellow solid).

[0197] Step 2: Compound 23a (3.8 g, 11.3 mmol) was added to 21 mL of anhydrous methanol, followed by the addition of 40% aqueous methylamine (4.5 g, 60.5 mmol) and stirring at room temperature for 1 h. After confirming completion of the reaction by LC-MS, the reaction mixture was concentrated to dryness. 40 mL of 1,4-dioxane was added, followed by the dropwise addition of 4 M hydrochloric acid and dioxane solution (15 mL, 64 mmol) and stirring at room temperature for 2 h. After confirming completion of the reaction by LC-MS, the filtrate was filtered and concentrated by rotary evaporation to give compound 23b (3.2 g, off-white solid).

[0198] 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. The mixture was heated to 80 °C and stirred for 2 h. After the reaction was confirmed to be complete by LC-MS, 200 mL of saturated sodium bicarbonate solution was added and stirred. The filtrate was filtered, concentrated by rotary evaporation, and purified by column chromatography to give compound 23 (1.8 g, white solid). LC-MS: ESI [M+H] + =426.5; 1 H NMR(400MHz,DMSO)δ 11.84(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).

[0199] Biological activity tests:

[0200] 1. Determination of PARP-1 / 2 / 5a enzymes

[0201] Materials: PARP1 protein (BPS, Catalog No. 80501), PARP2 protein (BPS, Catalog No. 80502), PARP5A protein (BPS, Catalog No. 80504), biotin-NAD+ (R&D, Catalog No. 6573), Strep-HRP (Thermo Pierce, Catalog No. 21127), NAD+ (TCl, Catalog No. D0919-5G), QuantaRed Enhanced HRP Substrate Kit (Thermo Pierce, Catalog No. 15159), histone (Active Motif, Catalog No. 81167), activated DNA (Genscript, Catalog No. L05182-01&02&03), anti-rabbit IgG, HRP-conjugated antibody (CST, Catalog No. 7074P2), anti-poly / mono-ADP-ribose (E6F6A) rabbit mAb (CST, Catalog No. 83732S), SuperSignal ELISA. Femto substrate (THERMO PIERCE, Cat. No. 37074) and reference compound AZD5305 purchased from MedChemExpress (MCE).

[0202] 1.1 Determination of PARP1 enzyme

[0203] 1.1.1 Preparation of buffer solutions: 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.

[0204] 1.1.2 Coating: A 50 ng / mL histone coating solution was prepared using 1x PBS, and 25 μL of the coating solution was transferred to a 384-well reaction plate, and the reaction plate was coated overnight at 4°C.

[0205] 1.1.3 Washing: After coating was completed, the coating solution was discarded, 50 μL of PBST was transferred to the 384-well reaction plate, and the plate was left for 5 minutes. The washing solution was discarded, the reaction plate was refilled, and the washing process was repeated three times to wash the reaction plate with PBST solution. Finally, the reaction plate was tapped dry and the next step of blocking was carried out.

[0206] 1.1.4 Blocking: 50 μL of blocking solution was transferred to a 384-well reaction plate and allowed to stand for 1 h.

[0207] Washing: After blocking was completed, discard the blocking solution, wash with PBST solution three times according to the method in step 2, and dry tap the reaction plate.

[0208] 1.1.5 1,000x compounds were prepared and 1 μL of compound was transferred to a 96-well plate containing 199 μL of reaction buffer, mixed thoroughly, and then 5 μL of the mixed compound was transferred to a 384-well reaction plate.

[0209] 1.1.6 25 / 10x PARP1-DNA solution was prepared using reaction buffer, and 10 μL of the PARP1-DNA solution was transferred into a 384-well reaction plate. For negative control wells, 10 μL of DNA solution was transferred, and the final concentration of PARP1 was 0.02 nM and the final concentration of DNA was 0.8 nM.

[0210] 1.1.7 25 / 10x NAD+ solution was prepared by using reaction buffer, and 10 μL of the NAD+ solution was transferred to a 384-well plate, resulting in a final NAD+ concentration of 3.5 μM, and the reaction plate was incubated at room temperature for 60 minutes.

[0211] 1.1.8 25 / 10x NAD+ solution was prepared by using reaction buffer, and 10 μL of the NAD+ solution was transferred to a 384-well plate, resulting in a final NAD+ concentration of 3.5 μM, and the reaction plate was incubated at room temperature for 60 minutes.

[0212] 1.1.9 Washing: After the reaction was completed, the reaction liquid was discarded. The reaction plate was washed three times with PBST solution according to the method in step 2, and the reaction plate was tapped dry.

[0213] 1.1.10 2,000-fold anti-poly / mono-ADP-ribose rabbit mAb was diluted in blocking solution, and 20 μL of diluted anti-poly / mono-ADP-ribose rabbit was added and incubated at room temperature for 1.5 h.

[0214] 1.1.11 Washing: Discard the anti-poly / mono-ADP-ribose rabbit, wash with PBST solution three times according to the method in step 2, and dry tap the reaction plate.

[0215] 1.1.12 2,000-fold dilution of anti-rabbit IgG, HRP-conjugated antibody was diluted with blocking solution, and 20 μL of diluted anti-rabbit IgG, HRP-conjugated antibody was added and incubated at room temperature for 1 hour.

[0216] 1.1.13 Washing: Discard the anti-rabbit IgG and HRP-conjugated antibody, wash with PBST solution three times according to the method in step 2, and dry tap the reaction plate.

[0217] 1.1.14 Color development: Femto-ECL Substrate A and Femto-ECL Substrate B were mixed in a 1:1 ratio and 25 μL of the mixture was transferred to a 384-well reaction plate.

[0218] 1.1.15 Reading: Relative light units (RLU) were read on the Envision.

[0219] 1.2 Determination of PARP2 enzyme

[0220] 1.2.1 Preparation of buffer solutions: PBST: 1x PBS, 0.05% Tween-20; Blocking solution: 1x PBS, 0.05% Tween-20, 5% BSA; Reaction buffer solution: 50 mM HEPES (pH 7.5), 0.002% Tween-20, 0.1% BSA, 100 mM NaCl, 2 mM DTT.

[0221] 1.2.2 Coating: A 100 ng / mL histone coating solution was prepared using 1x PBS, and 25 μL of the coating solution was transferred to a 384-well reaction plate, and the reaction plate was coated overnight at 4°C.

[0222] 1.2.3 Washing: After coating was completed, the coating solution was discarded, and 50 μL of PBST solution was transferred to the 384-well reaction plate, left for 5 minutes, the washing solution was discarded, the reaction plate was refilled, and the washing process was repeated three times to wash the reaction plate with PBST solution. Finally, the reaction plate was dry tapped and the next step of blocking was carried out.

[0223] 1.2.4 Blocking: 50 μL of blocking solution was transferred to a 384-well reaction plate and allowed to stand for 1 h.

[0224] 1.2.5 Washing: After the blocking was completed, discard the blocking solution, wash with PBST solution three times according to the method in step 2, and dry tap the reaction plate.

[0225] 1.2.6 Prepare a 25 / 10x PARP2 solution, then transfer 10 μL of the PARP2 solution to a 384-well reaction plate. For negative control wells, transfer 10 μL of reaction buffer solution instead, resulting in a final PARP2 concentration of 1.5 nM.

[0226] 1.2.7 Prepare 2,000x compounds, transfer 50 μL of compounds with eco-, add 19.95 μL of reaction buffer solution to the compounds, mix thoroughly, and transfer 5 μL of mixed compounds to a 384-well reaction plate.

[0227] 1.2.8 A 25 / 10x NAD+ solution was prepared and 10 μL of the biotin-NAD+ solution was transferred to a 384-well reaction plate, resulting in a final biotin-NAD+ concentration of 2 μM, and the reaction plate was incubated at room temperature for 60 minutes.

[0228] 1.2.9 Washing: After the reaction was completed, the reaction liquid was discarded. The reaction plate was washed with PBST solution three times according to the method in step 2, and the reaction plate was tapped dry.

[0229] 1.2.10 Stre-HRP solution was prepared by diluting with blocking solution, and 25 μL of Stre-HRP solution was transferred to the reaction plate and incubated at room temperature for 1 h, resulting in a final concentration of 0.1 μg / mL Stre-HRP.

[0230] 1.2.11 Washing: Discard the Stre-HRP solution, wash the reaction plate with PBST solution three times according to the method in step 2, and tap the reaction plate dry.

[0231] 1.2.12 Color development: Femto-ECL Substrate A, Femto-ECL Substrate B and QuantaRed ADHP were mixed in a ratio of 50:50:1. 25 μL of the mixture was transferred to a 384-well reaction plate and incubated at room temperature for 10 minutes, and 2.5 μL of QuantaRed Stop Solution was added.

[0232] 1.2.13 Reading: Fluorescence values ​​(Ex550 / Em620) were read on a Paradigm.

[0233] 1.3 Determination of PARP5A enzyme

[0234] 1.3.1 Preparation of buffer solutions: PBST: 1x PBS, 0.05% Tween-20; Blocking solution: 1x PBS, 0.05% Tween-20, 5% BSA; Reaction buffer solution: 50 mM HEPES (pH 7.5), 0.002% Tween-20, 0.1% BSA, 100 mM NaCl, 2 mM DTT.

[0235] 1.3.2 Coating: A 100 ng / mL histone coating solution was prepared using 1x PBS, and 25 μL of the coating solution was transferred to a 384-well reaction plate, and the reaction plate was coated overnight at 4°C.

[0236] 1.3.3 Washing: After coating was completed, the coating solution was discarded, 50 μL of PBST solution was transferred to the 384-well reaction plate, and left for 5 minutes. The washing solution was discarded, and the reaction plate was refilled with PBST solution. The washing process was repeated three times to wash the reaction plate with PBST solution. Finally, the reaction plate was dry tapped and the next step of blocking was carried out.

[0237] 1.3.4 Blocking: 50 μL of blocking solution was transferred to a 384-well reaction plate and allowed to stand for 1 h.

[0238] 1.3.5 Washing: After the blocking was completed, discard the blocking solution, wash with PBST solution three times according to the method in step 2, and dry tap the reaction plate.

[0239] 1.3.6 Prepare a 25 / 10x PARP5A solution, then transfer 10 μL of the PARP5A solution to a 384-well reaction plate. For negative control wells, transfer 10 μL of reaction buffer solution instead, resulting in a final concentration of 10 nM PARP5A.

[0240] 1.3.7 Prepare 2,000x compounds, transfer 50nL of compounds by eco-, add 19.95μL of reaction buffer solution to compounds, mix thoroughly, and transfer 5μL of mixed compounds to a 384-well reaction plate.

[0241] 1.3.8 A 25 / 10x NAD+ solution was prepared and 10 μL of the biotin-NAD+ solution was transferred to a 384-well reaction plate, resulting in a final biotin-NAD+ concentration of 2 μM, and incubated at room temperature for 60 minutes.

[0242] 1.3.9 Washing: After the reaction was completed, the reaction liquid was discarded. The reaction plate was washed three times with PBST solution according to the method in step 2, and the reaction plate was tapped dry.

[0243] 1.3.10 Stre-HRP solution was prepared by diluting with blocking solution, and 25 μL of Stre-HRP solution was transferred to the reaction plate and incubated at room temperature for 1 h, resulting in a final concentration of 0.1 μg / mL Stre-HRP.

[0244] 1.3.11 Washing: Discard the Stre-HRP solution, wash with PBST solution three times according to the method in step 2, and dry tap the reaction plate.

[0245] 1.3.12 Color development: Femto-ECL Substrate A, Femto-ECL Substrate B and QuantaRed ADHP were mixed in a ratio of 50:50:1. 25 μL of the mixture was transferred to a 384-well reaction plate and incubated at room temperature for 10 minutes, and 2.5 μL of QuantaRed Stop Solution was added.

[0246] 1.3.13 Reading: Fluorescence values ​​(Ex550 / Em620) were read on a Paradigm.

[0247] The test results are shown in Table 2: [Table 2]

[0248] Conclusion: The compounds of the present invention exhibited effective inhibitory effects on PARP1, but showed weak inhibition on PARP2 / 5a, indicating that the compounds of the present invention selectively inhibit PARP-1.

[0249] 2. Cell Antiproliferative Activity Assay

[0250] BRCA-mutated MDA-MB-436 cells were cultured in DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin. The cells were incubated at 37°C in a 5% CO2 incubator. When the cells reached 80% confluence, they were harvested, centrifuged at 300 g for 10 minutes, and then plated in a 96-well plate at a density of 1,200 cells per well. After 24 h, PARPi (0, 0.01, 0.1, 1, 10, 100, and 1000 nM) was added at various final concentrations, and the cells were further cultured for 72 h. The cells were subjected to a fluid exchange treatment (adding PARPi at the same final concentration again) and then cultured for 96 h. The 96-well plate was removed, and the OD value was measured at 450 nm using the CCK8 method. The cell inhibition rate was then calculated using the following formula: Inhibition rate (%) = 1 - (mean OD value of the treatment group - mean OD value of the blank group) / (mean OD value of the control group - mean OD value of the blank group) * 100%.

[0251] BRCA wild-type DLD-1 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin. The cells were incubated at 37°C in a 5% CO2 incubator. When the cells reached 80% confluence, they were harvested, centrifuged at 300 g for 10 minutes, and plated at a density of 1,000 cells per well in a 96-well plate. After 24 h, PARPi (0.1 and 10 μM) with varying final concentrations was added, and the cells were further cultured for 72 h. The cells were subjected to a fluid exchange treatment (adding PARPi at the same final concentration again) and then cultured for 96 h. The 96-well plate was removed, and the OD value was measured at a wavelength of 450 nm using the CCK8 method. The cell inhibition rate was calculated:

[0252] Inhibition rate (%) = 1 - (mean OD value of treatment group - mean OD value of blank group) / (mean OD value of control group - mean OD value of blank group) * 100%.

[0253] The test results are shown in Table 3 below: [Table 3]

[0254] Conclusion: The compounds of the present invention exhibited effective inhibitory effects in BRCA-mutated MDA-MB-436 cells, but showed obvious inhibitory effects in BRCA wild-type DLD-1 cells, indicating that the compounds of the present invention specifically inhibit tumor cells with homologous recombination repair deficiency.

[0255] 3. Pharmacokinetic evaluation of compounds in Balb / c mice

[0256] Experimental objective: To learn the pharmacokinetic profile of the compound.

[0257] Experimental basis: Technical Guidelines for Non-clinical Pharmacokinetic Studies of Chemical Drugs, 2014.

[0258] Experimental scheme: The pharmacokinetic profile of the compound was determined by administering the compound intravenously (1 mg kg -1 ) and oral (1 mg kg -1 ) was administered.

[0259] Sample preparation: Approximately 0.2 mg of compound was dissolved in 10 μL of DMSO, and then sodium chloride injection was added to give 0.1 mg mL of compound for administration. -1 A solution was made.

[0260] Sample collection: Six male Balb / c mice (Chengdu Dossy Experimental Animal Co., Ltd. License number: SCXK(C)2020-030) were used. 1 mg kg -1 Three mice were intravenously (IV) administered the compound at a dose of 0.05 mL, and the other three mice were orally (PO) administered the compound at the same dose. Approximately 0.05 mL of blood samples were collected 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 24 hours, and 48 hours after administration. The collected blood samples were centrifuged at 3,500 rpm for 15 minutes, and the supernatant plasma was collected and stored at -40°C until assay. The plasma concentrations of the drug were quantitatively analyzed using LC-MS / MS, and the time to peak concentration (Cmax), area under the concentration-time curve (AUC(0-t)), and half-life (T 1 / 2 Pharmacokinetic parameters including serotonin (S), clearance (CL), volume of distribution at steady state (Vdss) and bioavailability (F) were calculated.

[0261] The pharmacokinetic evaluation results are shown in Table 4 below: [Table 4]

[0262] Conclusion: The compounds of the present invention exhibit favorable pharmacokinetic properties in Balb / c mice, including good oral bioavailability, exposure levels, half-lives, and clearance rates. max was superior to the Cmax of compounds 1, 2, 3, 15 and 21 as well as the reference compound 5305.

[0263] 4. Pharmacokinetic evaluation of compounds in SD rats

[0264] Experimental objective: To learn the pharmacokinetic profile of the compound.

[0265] Experimental basis: Technical Guidelines for Non-clinical Pharmacokinetic Studies of Chemical Drugs, 2014.

[0266] Experimental scheme: The pharmacokinetic profile of the compound was investigated by oral intravenous administration in SD rats.

[0267] Experimental procedure: The compound, a small amount of DMSO, and sodium chloride injection were mixed to prepare the final solution for administration. The compound was orally administered to six male SD rats. Approximately 0.1 mL of blood samples were collected 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, and 24 hours after administration. The blood samples were centrifuged at 3,500 rpm for 15 minutes, and the supernatant plasma was collected. 5 μL of plasma was transferred to an EP tube and diluted to 20 ng mL -1 Add 100 μL of acetonitrile-precipitated protein containing the internal standard SAHA, vortex for 30 seconds, and centrifuge at 13,000 rpm for 15 minutes. Collect the supernatant and transfer it to a sample vial for assay. Standard curve range: 10–10,000 ng mL -1 .

[0268] The pharmacokinetic evaluation results are shown in Table 5 below: [Table 5]

[0269] Conclusion: The compounds of the present invention exhibit favorable pharmacokinetic properties in SD rats, including good oral bioavailability, exposure level, half-life, and clearance rate. max was superior to that of the reference compounds AZD5305, Compound 1 and Compound 21.

[0270] 5. In vivo pharmacodynamic study of compounds 1 and 6 in a subcutaneous xenograft tumor model of breast cancer MDA-MB-436 in nude mice

[0271] (1) Main equipment and devices

[0272] CO2 cell culture incubator: Yamato, IP610, safety cabinet: Suzhou Antai, BSC-1304IIA2, tabletop centrifuge: Thermo Scientific, SORVALL ST 16. Digital inverted microscope: Olympus, CKX3-SLP, constant temperature water bath: Shanghai Yuejin Medical Equipment Co., Ltd., HSW-420, cell counting chamber: Shanghai Qiujing Biochemical Reagent Instrument Co., Ltd., XB.K.25, liquid nitrogen tank: Thermo, CY50935-70, refrigerator: Haier, BCD-601WDGX, medical low-temperature refrigerator: Thermo, ULTS1651.

[0273] Water purifier: Millipore, F7PNO9748, vertical autoclave: Yamato, DKN812C, 1mL disposable sterile syringe: Shanghai Kindly Enterprise Development Group Co., Ltd., 1mL disposable sterile insulin syringe: BD, scale: Shanghai Hengping Instrument Co., Ltd., JY2002, analytical balance: SARTORIUS, BCE95I-1CEU.

[0274] Thermo-hygrometer: Wuqiang Hygrothermograph Manufacturing Center, LZJZ30260102, mini centrifuge: Thermo, SORVALL LEGEND MICRO 17 Centrifuge, mini mixer: Yeasen, ES-VM25, metal bath: SCILOGEX, SCL120-S.

[0275] (2) Main software and data processing systems

[0276] Graphpad Prism (version 6.0) from Graphpad Software, Inc. was used to assemble, quantify, and generate bar graphs.

[0277] (3) Experimental animals

[0278] Strain: NOD / SCID, Grade: SPF, Source: Beijing Vitron River Bio-technology Co., Ltd.

[0279] (4) Formation method

[0280] Tumor cell culture and preparation: Conventional tumor cell lines were passaged and cultured in culture medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin. When cells reached 80% confluence, they were harvested and centrifuged at 300 g for 10 minutes. The cells were washed three times with pre-chilled PBS, centrifuged at 300 g for 10 minutes, harvested, resuspended in PBS, counted in a hemocytometer, and the cell concentration was adjusted. After cell harvest, the cell suspension was pre-chilled on ice. 100 μL of the cell suspension was administered by subcutaneous injection into the dorsal flank of mice, and tumors with a volume of 200–300 mm were cultured. 3 Grouped treatments were initiated when the

[0281] (5) Observation and detection indicators

[0282] General Observations: Animal Observations: All live experimental animals observed; Experimental Time: Twice a day; Experimental Items: Including but not limited to general behavior, behavioral state, eyes, mouth, nose and mouth, ears, hair, feces, urine, genitals and other toxic symptoms; If any of the above are observed, provide a detailed description.

[0283] (6) Weight

[0284] Animal assay: All live experimental animals scheduled for the assay. Assay time: All animals were weighed before modeling for the purpose of test grouping. After modeling, mice were measured every two days, and changes in mouse weight were recorded in real time. Valid criteria: tumor volume (TV), relative tumor volume (RTV), and relative tumor growth rate of the subcutaneously implanted tumor model. The major and minor diameters of the tumor were measured with a caliper every two days to dynamically observe the antitumor effect of the tested drug. The formula for calculating tumor volume (TV) is TV (mm 3 )=a×b2×0.5, where a and b are the major and minor diameters, respectively.

[0285] Based on the measurement results, the relative tumor volume (RTV) was calculated according to the formula: RTV = Vt / V0, where V0 is the tumor volume (TV) measured for each group at the start of treatment (i.e., d0), and Vt is the tumor volume measured for that group at each subsequent time point. The evaluation index for antitumor activity is the relative tumor growth rate T / C (%), which is calculated as follows: T / C(%)=TRTV / CRTV×100% Where TRTV is the relative tumor volume of the treatment group, and CRTV is the relative tumor volume of the negative control group, as follows: Treatment evaluation criteria: T / C (%)>60 is considered ineffective, and T / C (%)≦60 and P<0.05 (by statistical analysis) is considered effective. Measurement of tumor weight and tumor inhibition rate (%): At the end of the treatment period, the animals were euthanized, the tumor mass was dissected and divided, and the tumor was weighed and photographed. The tumor inhibition rate (%) was calculated using the following formula:

[0286] Tumor inhibition rate (tumor growth inhibition rate, %) = [(mean tumor weight (g) of the negative control group - mean tumor weight (g) of the treatment group)] / mean tumor weight (%) of the negative control group × 100%.

[0287] Effective evaluation criteria: A tumor inhibition rate (i.e., tumor growth inhibition rate) of <40% was considered ineffective, and a tumor growth inhibition rate of ≥40% and P<0.05 (by statistical analysis) was considered effective. Overall evaluation criteria for the subcutaneous tumor model: If the above two effective evaluation criteria (relative tumor growth rate and tumor inhibition rate) met the effectiveness criteria, it was considered effective.

[0288] (7) Data acquisition and analysis

[0289] All raw data within the facility were collected manually or using a data acquisition system according to the experimental scheme and relevant institutional guidelines. The manually collected data were transferred to software such as Excel for analysis and reporting. Data from each group was quantitative, and experimental data for each group of animals in tables were presented as the mean ± standard deviation (mean ± SD). In figures, experimental data were presented as the mean ± standard error of the mean (mean ± SEM). Statistical analyses, including t-tests and survival analyses, were performed using software such as Excel and GraphPad Prism.

[0290] Experimental Procedure: A total of 42 female NOD / SCID mice were used in the experiment and randomly divided into seven groups: a blank control group, three Compound 6 treatment groups (0.03, 0.1, and 0.3 mg / kg), a reference compound group (AZD5305 0.1 mg / kg), a Compound 1 group (0.1 mg / kg), and an olaparib group (100 mg / kg). Each group consisted of six mice, all of which were orally administered once daily. Mice were weighed every two days, and tumor length and width were measured with a caliper. After 24 days of treatment, tumor-bearing mice were anesthetized and euthanized. Tumor tissues were dissected, weighed, and photographed to calculate the tumor inhibition rate.

[0291] Conclusion: The experimental results are shown in Figure 1 and Table 6. After cell inoculation, each treatment group was administered continuously for 24 days. Compared with the blank control group, all treatment groups (AZD5305 group: 0.1 mg / kg, Compound 1 group: 0.1 mg / kg, Olaparib group: 100 mg / kg, and Compound 6 group: 0.03, 0.1, and 0.3 mg / kg) showed effective tumor growth inhibition in a NOD / SCID mouse subcutaneous tumor model of the human breast cancer cell line MM436. The tumor inhibition rates for each treatment group were 80.79%, 82.18%, 62.19%, 49.77%, 86.73%, and 95.83%, respectively. The relative tumor growth rates (T / C%) were 15.2%, 17.0%, 33.4%, 40.8%, 12.6%, and 3.8%, respectively. Among all treatment groups, the compound 6 group (0.3 mg / kg) showed the best tumor inhibition effect, with complete tumor regression observed in 2 out of 6 mice (2 / 6) at the end of treatment. The tumor inhibition effect of the compound 6 group (0.1 mg / kg) was superior to that of the reference compounds AZD5305 and compound 1 at the same dose, and was significantly better than that of the reference drug olaparib group (100 mg / kg) (P<0.01). Furthermore, there was a significant dose-dependent difference between the low, medium, and high dose groups of compound 6 (P<0.001, P<0.01).

[0292] [Table 6]

[0293] 6. Challenge study of compounds 1, 6, and 21 in a subcutaneous xenograft tumor model of breast cancer MDA-MB-436 in nude mice

[0294] Experimental procedure: 15 female NOD / SCID mice were used in the experiment and randomly divided into three groups: Compound 1 (1 mg / kg), Compound 6 (1 mg / kg), and Compound 21 (1 mg / kg). Each group consisted of 5 mice, and the mice were orally administered once a day. The mice were weighed every 2 days, and the length and width of the tumor were measured with a vernier caliper.

[0295] Conclusion: The experimental results are shown in Figure 2. Compounds 1, 6, and 21 of the present invention still exhibit excellent antitumor efficacy in large tumor models, and can cause complete tumor regression. Compound 6 exhibits a faster tumor regression rate compared to compounds 1 and 21, indicating that at the same dose, compound 6 has stronger in vivo antitumor efficacy than compounds 1 and 21.

[0296] 7. In vivo pharmacodynamics study of compound 6 in a PDX gastric cancer mouse model

[0297] Experimental Procedure: A stable tumor-bearing mouse model was established using tumor tissue from a gastric cancer patient. In this study, tumor fragments from mice that had been stably passaged three times were used for subcutaneous inoculation. Eight groups were divided for the experiment, with six mice in each group. The negative control group (Control) received 0.2 mL of saline per mouse per dose and was orally administered once daily. The carboplatin control group (Carboplatin, Car) received 20 mg / kg at 0.2 mL per mouse per dose and was administered intraperitoneally once every seven days. Test compound 6: 0.3, 1, and 3 mg / kg, 0.2 mL per mouse per dose and was orally administered once daily. The combination group received compound 6 plus carboplatin.

[0298] Details of the dosing regimen and groupings are shown in Table 7. [Table 7]

[0299] Conclusion: The experimental results are shown in Figure 3. When combined with carboplatin, different doses of compound 6 exhibited a dose-dependent inhibitory effect on gastric cancer PDX models. In particular, the 3 mg / kg group of compound 6 combined with carboplatin significantly inhibited tumor growth.

[0300] 8. In vivo pharmacodynamic study of compound 6 in a subcutaneous xenograft tumor model of breast cancer SUM14PT in nude mice

[0301] Experimental Procedure: In this study, a mouse model was established by subcutaneous inoculation with SUM14PT tumor cells. Eight groups were divided for the experiment, each with six mice. The negative control group (Control) received 0.2 mL of saline per mouse per dose, administered orally once daily. The carboplatin control group (Carboplatin, Car) received 20 mg / kg at 0.2 mL per mouse per dose, administered by intraperitoneal injection once every seven days. The olaparib control group: 100 mg / kg, administered orally at 0.2 mL per mouse per dose, once daily. The test compound 6 groups (0.3 and 1 mg / kg): received 0.2 mL per mouse per dose, administered orally once daily. The combination groups received either olaparib or compound 6 plus carboplatin.

[0302] Details of the dosing regimen and groupings are shown in Table 8. [Table 8]

[0303] Conclusion: The experimental results are shown in Figure 4. Compound 6, administered orally once daily at doses of 0.3 and 1 mg / kg, showed better antitumor efficacy than olaparib administered orally at 100 mg / kg once daily as monotherapy. Furthermore, compound 6 administered orally at 1 mg / kg once daily in combination with carboplatin showed better therapeutic efficacy than olaparib administered orally at 100 mg / kg once daily in combination with carboplatin.

Claims

1. A compound represented by Formula II, or a pharmaceutically acceptable form thereof, wherein said Formula II has the following structure, and said pharmaceutically acceptable form is selected from the group consisting of a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug: 【Chemistry 1】 During the ceremony, 【Chemistry 2】 represents a single bond or a double bond, R １ But C １－４ deuterated alkyl; X １ is N or C(R ５ａ ) and X ２ is N or C(R ５ｂ ) and X ３ is N or C(R ５ｃ ) and X １ , X ２ and X ３ is selected from N, R ２ａ and R ２ｂ are independently selected from hydrogen, deuterium, methyl, or deuteromethyl; R ３ But deuterium, fluorine, C １－４ Alkyl or C １－４ deuterated alkyl; R ３ａ But hydrogen, deuterium, fluorine, hydroxy, cyano, C １－４ Alkyl, C １－４ Fluoroalkyl or C １－４ alkoxy; R ４ But hydrogen, halogen, cyano, C １－４ Alkyl, C １－４ Deuterated alkyl, C １－４ Fluoroalkyl or -CONHR ７ is selected from R ５ａ , R ５ａ and R ５ｃ However, hydrogen, fluorine, chlorine, C １－４ Alkyl, C １－４ Deuterated alkyl, C １－４ Fluoroalkyl, C １－４ Alkoxy or C １－４ fluoroalkoxy; R ６ However, hydrogen, fluorine, chlorine, C １－４ Alkyl, C １－４ Fluoroalkyl or C １－４ deuterated alkyl; R ７ But hydrogen, C １－４ Alkyl, C １－４ Deuterated alkyl, C １－４ selected from fluoroalkyl or 3- to 6-membered cycloalkyl; X ５ is nitrogen or C(R ９ａ ) and X ６ is nitrogen or C(R ９ｂ ) and X ７ is nitrogen or C(R ９ｃ ) and X ８ is nitrogen or C(R ９ｄ ) are selected from R ９ａ , R ９ｂ , R ９ｃ and R ９ｄ But hydrogen, fluorine, chlorine, cyano, C １－４ Alkyl, C １－４ Deuterated alkyl, C １－４ Fluoroalkyl, C １－４ Alkoxy or C １－４ fluoroalkoxy; n1 is an integer selected from 0 to 8; n3 is independently selected from 0 or 1.

2. R １ 2. The compound according to claim 1, wherein is selected from deuterated methyl or deuterated methyl.

3. R １ But, -CD ２ CD ３ or -CH ２ CD ３ 3. The compound according to claim 2, characterized in that it is selected from:

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

5. R ３ 5. The compound according to claim 4, characterized in that is selected from deuterium or methyl. 【Request Item 6】 【Chemistry 3】 is a double bond, R ３ａ does not exist, 【Chemistry 4】 is a single bond, R ３ａ Compounds according to any one of claims 1 to 5, characterized in that is selected from hydrogen or deuterium.

7. R ４ However, fluorine, chlorine, cyano, C １－４ Alkyl, C １－４ Deuterated alkyl, C １－４ Fluoroalkyl, C １－４ Alkylaminocarbonyl, C １－４ Deuterated alkylaminocarbonyl, C １－４ selected from fluoroalkylaminocarbonyl or 3- to 6-membered cycloalkylaminocarbonyl; Preferably, R ４ Compounds according to any one of claims 1 to 6, characterized in that is selected from fluorine, cyano, methylaminocarbonyl, deuterated methylaminocarbonyl or cyclopropylaminocarbonyl.

8. R ６ Compounds according to any one of claims 1 to 7, characterized in that is selected from hydrogen, fluorine, chlorine or methyl.

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

10. The compound of any one of claims 1 to 9, characterized in that it has a structure represented by formula VI-I: 【Transformation 5】 During the ceremony, R ６ is selected from hydrogen or fluorine, R ９ａ But hydrogen, fluorine, chlorine, cyano, C １－４ Alkyl, C １－４ Deuterated alkyl or C １－４ fluoroalkyl; n1 is an integer selected from 0 to 6.

11. The compound of any one of claims 1 to 10, characterized in that it has the structure of formula VI-3, or a pharmaceutically acceptable form thereof: 【Transformation 6】 During the ceremony, R ６ is selected from hydrogen or fluorine, R ９ａ But hydrogen, fluorine, chlorine, cyano, C １－４ Alkyl, C １－４ Deuterated alkyl or C １－４ fluoroalkyl; n1 is an integer selected from 0 to 6.

12. The compound is 【Transformation 7】 Compounds according to any one of claims 1 to 11, characterized in that they are selected from:

13. The compound is 【Transformation 8】 Compounds according to any one of claims 1 to 11, characterized in that they are selected from:

14. The compound is 【Chemistry 9】 Compounds according to any one of claims 1 to 11, characterized in that they are selected from:

15. A pharmaceutical composition comprising, as an active ingredient, a compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, and a pharmaceutically acceptable carrier.

16. A compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labelled compound, metabolite or prodrug thereof, and a pharmaceutical composition according to claim 15, intended for use in the manufacture of a medicament for the prevention and / or treatment of a PARP1 enzyme related disease.

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

18. 18. The use according to claim 17, wherein the neoplastic disorder is characterized by a deficiency in the HR-dependent repair pathway for DNA DSBs.

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

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

21. Use according to any one of claims 17 to 20, characterized in that the neoplastic disorder comprises breast cancer, ovarian cancer, primary peritoneal cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer, glioblastoma or lung cancer.

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