Compounds having PARP1 inhibitory activity and uses thereof

Abstract

Disclosed are compounds having poly ADP-ribose polymerase inhibitory activity, pharmaceutical compositions and uses thereof. In particular to compounds of formula (I), and also to pharmaceutical compositions comprising these compounds and to the use of these compounds for the preparation of medicaments for the prevention and / or treatment of diseases, in particular diseases ameliorated by the inhibition of PARP1. # imgabs0 #

Description

Compounds with PARP1 inhibitory activity and uses thereof Technical Field

[0001] The present invention relates to a class of compounds capable of inhibiting poly ADP-ribose polymerase 1 (PARP1) activity and uses thereof, and in particular to a class of quinolone compounds, pharmaceutical compositions comprising the compounds, and uses thereof in drugs for treating diseases improved by inhibiting PARP1, particularly tumor diseases. Background Art

[0002] Poly (ADP-ribose) polymerases (PARPs) are an emerging family of enzymes that catalyze the transfer of ADP-ribose to target proteins (poly (ADP-ribosylation)). At least 18 PARP family members are encoded by different genes and share homology in the conserved catalytic domain (Morales et al, Critical Reviews TM in Eukaryotic Gene Expression 24.1, 2014). PARP1, short for poly(ADP-ribose) polymerase 1, is an abundant nuclear protein (Murai et al, Cancer Research 72.21, 2012). PARP1 catalyzes the transfer of ADP-ribose residues from NAD+ to target substrate proteins or nucleic acids, constructing a poly(ADP-ribose) (PAR) chain that is added to downstream target proteins. This post-translational modification is called PARylation. PARPs play an important role in several cellular processes, including cell proliferation and cell death (Murai et al, Cancer Research 72.21, 2012). The main function of PARP is to participate in DNA damage repair. Single-strand breaks (SSBs) are the most common type of damage and can be converted into potentially disruptive and lethal double-strand breaks (DSBs). PARP1 binds to damaged DNA at single-strand breaks (SSBs) and other DNA damage. This event causes a series of conformational changes in the structure of PARP1, thereby activating its catalytic function (Lord et al, Science 355.6330, 2017).

[0003] BRCA1 and BRCA2 proteins are crucial for the repair of double-stranded DNA breaks (DSBs) through a process called homologous recombination repair (HRR), a form of DNA repair that utilizes homologous DNA sequences to guide repair at the site of a DSB (Lord et al, Science 355:6330, 2017). HRR is normally a "conservative" mechanism because it restores the original DNA sequence at the site of DNA damage. When cells are deficient in HRR, whether driven by defects in BRCA1, BRCA2, or other pathway components, non-conservative forms of DNA repair, such as non-homologous end joining (NHEJ), predominate.

[0004] PARP inhibitors exert their anti-cancer effects by blocking DNA damage repair in highly mutated cancer cells, resulting in "toxic damage" that causes cell death due to homologous recombination repair (HRR) deficiency. Healthy cells contain multiple signaling pathways for DNA repair, so inhibiting only PARP is not very toxic. However, certain tumor cells, due to specific gene mutations such as BRCA, can disrupt other DNA repair pathways, making them dependent on PARP-1 and therefore particularly sensitive to PARP inhibitors. This is why patients with ovarian and breast cancers carrying BRCA mutations are more likely to benefit from PARP inhibitors. PARP2 levels are low, accounting for only 5% to 10% of total PARP activity. Knocking out PARP1 significantly reduces PARP activity compared to knocking out PARP2 (<10%) (Yélamos et al, The EMBO journal 25.18, 2006). Knocking out PARP1 blocks the inhibitory activity of olaparib on PARP and also eliminates the cell proliferation inhibitory effect of olaparib (Murai et al, Cancer research, 2012). These data indicate that PARP1 is the key to determining the efficacy of PARPi. Literature reports indicate that intact PARP2 in the bone marrow is required for mouse survival; PARP2 deficiency leads to reduced RBC, WBC, and BM cell numbers (Farrés et al, Blood, The Journal of the American Society of Hematology 122.1, 2013); and compared to PARP1 knockout, PARP2 knockout reduces T cell and RBC numbers, whereas PARP1 knockout has no significant effect on T cell (Yélamos et al, Blood, The EMBO journal 25.18, 2006) or RBC numbers (Farrés et al, Cell Death & Differentiation 22.7, 2015). Therefore, PARP1 inhibition is the primary source of drug efficacy, while PARP2 inhibition is the primary source of toxicity. The development of highly selective PARP1 / 2 inhibitors could significantly reduce the toxicity caused by PARP2 without significantly reducing drug efficacy.

[0005] In summary, there is an urgent need in this field to develop PARP inhibitors with high efficacy and good safety, especially inhibitors with high selectivity for PARP1.

[0006] Summary of the Invention

[0007] The present invention relates to a compound represented by general formula (I), or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof.

[0008] in,

[0009] X 1 、X 2 Each independently selected from N and C(H), C(OCH3);

[0010] X 3 Selected from N and CR 10 , where R 10 is hydrogen or halogen; preferably, R 10 is hydrogen or fluorine;

[0011] R 1 is hydrogen, C1-C6 alkoxy, C1-C6 alkyl which is unsubstituted or substituted by halogen, or C3-C6 cycloalkyl which is unsubstituted or substituted by halogen; preferably hydrogen, C1-C4 alkoxy, C1-C4 alkyl which is unsubstituted or substituted by halogen, or C3-C6 cycloalkyl which is unsubstituted or substituted by halogen; more preferably hydrogen, C1-C2 alkoxy, C1-C4 alkyl, C3-C6 cycloalkyl, or C1-C4 fluoroalkyl; further preferably hydrogen, methoxy, methyl, ethyl, 1-fluoroethyl, difluoromethyl, or trifluoromethyl;

[0012] R 2 is hydrogen, halogen or C1-C6 alkyl; preferably hydrogen, halogen or C1-C4 alkyl; more preferably hydrogen or fluorine;

[0013] R 3 、R 4 、R 4‘ 、R 5 、R 6 、R 7 Each independently is hydrogen or C1-C6 alkyl; preferably hydrogen or C1-C4 alkyl;

[0014] R 8 is selected from hydrogen, halogen, unsubstituted or halogen-substituted C1-C6 alkyl; preferably, R 8 is selected from hydrogen, halogen, unsubstituted or halogen-substituted C1-C4 alkyl; more preferably, R 8 is selected from hydrogen, halogen, C1-C4 alkyl and C1-C4 fluoroalkyl;

[0015] R 9 It is hydrogen, C3-C6 cycloalkyl or C1-C6 alkyl; preferably it is hydrogen, C3-C6 cycloalkyl or C1-C4 alkyl; more preferably it is hydrogen or C1-C4 alkyl;

[0016] m is 1 or 2; preferably 1;

[0017] The conditions are:

[0018] When X 1 is N, and X 2 and X 3 When it is C(H), then R 3 、R 4 、R 4‘ 、R 5 、R 6 、R 7 Not simultaneously hydrogen;

[0019] When R 2 When it is H, then R 3 、R 4 、R 4‘ 、R 5 、R 6 、R 7 are not simultaneously hydrogen; and

[0020] When R 3 is H and X 3 When it is C(H), then R 2 is F, and R 4 、R 4‘ 、R 5 、R 6 、R 7 Not hydrogen at the same time,

[0021] The compounds described do not include the following compounds:

[0022] Preferably, the condition is that R 3 、R 4 、R 4‘ 、R 5 、R 6 、R 7 Not hydrogen at the same time.

[0023] In a preferred embodiment, X 3 is N, and other substituents are as described above.

[0024] In a preferred embodiment, X 3 CR 10 , R 10 is hydrogen or halogen, and other substituents are as described above. More preferably, R 10 It is H, F or Cl.

[0025] In a preferred embodiment, X 1 is selected from N, C(H), C(OCH3); other substituents are as described above. More preferably, X 1is selected from N and C(H).

[0026] In a preferred embodiment, X 2 is N or C(H); other substituents are as described above.

[0027] In a preferred embodiment, X 2 is C(H) or C(OCH3); other substituents are as described above.

[0028] In a preferred embodiment, X 2 is C(H); other substituents are as described above.

[0029] In a preferred embodiment, R 2 is hydrogen, F, Cl or C1-C4 alkyl; other substituents are as described above. More preferably, R 2 is hydrogen, F or methyl; more preferably, R 2 It's F.

[0030] In a preferred embodiment, R 2 It's F, X 2 is C(H); other substituents are as described above.

[0031] In a preferred embodiment, X 1 is selected from N, C(H) and C(OCH3); X 2 is selected from C(H) and C(OCH3); X 3 N, R 2 is hydrogen or F, and other substituents are as described above. 2 Selected from C(H).

[0032] Preferably, in the structure of formula (I) Partially selected from the following structures:

[0033] More preferably, selected from

[0034] More preferably, selected from

[0035] The present invention relates to a compound represented by general formula (II), or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof.

[0036] Preferably, in the compound represented by (I) or (II), R 2 is hydrogen, F, Cl or C1-C4 alkyl; more preferably, R 2 is hydrogen, F or methyl. Further preferably, R 2 It's F.

[0037] In a preferred embodiment, in the compound represented by (I) or (II), R 8 is selected from hydrogen and halogen, more preferably, R 8 Selected from hydrogen, F, Cl.

[0038] Preferably, the compound represented by (I) has a structure represented by formula (I-1) to formula (I-8) or a racemate thereof:

[0039] More preferably, the compound of formula (I) or (II) is selected from the following specific compounds:

[0040] On the other hand, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate or prodrug thereof, and a pharmaceutically acceptable carrier.

[0041] In certain embodiments of the pharmaceutical composition, the pharmaceutical composition is formulated for intravenous administration, intramuscular administration, oral administration, rectal administration, inhalation administration, nasal administration, topical administration, ophthalmic administration, or ear administration. In other embodiments of the pharmaceutical composition, the pharmaceutical composition is a tablet, pill, capsule, liquid, inhalant, nasal spray solution, suppository, solution, emulsion, ointment, eye drops, or ear drops. In other embodiments of the pharmaceutical composition, it further comprises one or more additional therapeutic agents.

[0042] On the other hand, the present invention provides the use of a compound of formula (I), or its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, crystal forms, solvates, hydrates or prodrugs, or the pharmaceutical composition thereof in the preparation of a medicament for preventing, treating or ameliorating diseases by inhibiting PARP1.

[0043] On the other hand, the present invention provides a method for preventing, treating or ameliorating a disease by inhibiting PARP1, which comprises administering to an individual in need of such treatment an effective amount of a compound of formula (I), or its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, crystal forms, solvates, hydrates or prodrugs, or said pharmaceutical composition.

[0044] In some embodiments of the present invention, the disease includes but is not limited to cancer.

[0045] In some embodiments of the present invention, the cancer genome is of a type deficient in homologous recombination repair.

[0046] In some embodiments of the invention, the cancer is dependent on a pathway that is deficient in homologous recombination repair of DNA double-strand damage.

[0047] In some embodiments of the present invention, the cancer comprises one or more cancer cells that lack the ability to repair DNA double-strand breaks by homologous recombination relative to normal cells.

[0048] In some embodiments of the invention, the cancer comprises one or more cancer cells that lack BRCA1 or BRCA2 or that have a BRCA1 or BRCA2 mutation.

[0049] In some embodiments of the present invention, the cancer includes, but is not limited to, a malignant tumor, such as any one of ovarian cancer, breast cancer, fallopian tube cancer, endometrial cancer, peritoneal cancer, gastric cancer, colon cancer, bladder cancer, pancreatic cancer, biliary tract cancer, osteosarcoma, cervical cancer, head and neck tumors, germ cell and embryonal cancers, esophageal cancer, malignant glioma, Ewing sarcoma, pancreatic cancer, melanoma, bile duct cancer, prostate cancer, small cell lung cancer, non-small cell lung cancer, lymphoma, and blood cancer.

[0050] Terminology

[0051] In the present invention, unless otherwise specified, the terms used in the present invention have the meanings defined below. Terms not clearly defined in the present invention have the general meanings generally understood by those skilled in the art.

[0052] As used herein, the term "halogen" or "halo" refers to fluorine, chlorine, bromine, and iodine.

[0053] As used herein, the terms "optional," "optionally," or "optionally" mean that the subsequently described substitution pattern, event, or circumstance may or may not occur, and that the description includes instances where the substitution pattern occurs as well as instances where the substitution pattern does not occur. For example, "optionally substituted alkyl" includes "unsubstituted alkyl" and "substituted alkyl" as defined herein. It will be understood by those skilled in the art that, for any group containing one or more substituents, the group does not include any substitution pattern that is sterically impractical, chemically incorrect, synthetically infeasible, and / or inherently unstable.

[0054] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological effects and properties of the compounds of the present invention and is not biologically or otherwise undesirable. Non-limiting examples of such salts include non-toxic, inorganic or organic base or acid addition salts of the compounds of the present invention. In many cases, the compounds of the present invention are capable of forming acid salts and / or base salts due to the presence of amino and / or carboxyl groups or groups similar thereto. Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like; ammonium, potassium, sodium, calcium, and magnesium salts are particularly preferred. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, basic ion exchange resins, and the like, particularly isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. Pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound (basic or acidic moiety) by conventional chemical methods. Generally, the salts can be prepared by reacting the free acid form of the compound with a stoichiometric amount of an appropriate base (e.g., hydroxide, carbonate, bicarbonate, etc., of Na, Ca, Mg, or K) or by reacting the free base form of the compound with a stoichiometric amount of an appropriate acid. Such reactions are typically carried out in water or an organic solvent, or a mixture of the two. In general, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred where practicable. Other suitable salts can be found in Remington's Pharmaceutical Sciences, 20th ed., Mack Publishing Company, Easton, Pa., (1985), which is incorporated herein by reference.

[0055] As used herein, the term "pharmaceutically acceptable excipient" includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drugs, drug stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavorings, dyes, the like, and combinations thereof, which are well known to those of ordinary skill in the art (see, for example, Remington's Pharmaceutical Sciences, 18th ed., Mack Printing Company, 1990, pp. 1289-1329, incorporated herein by reference). Unless any conventional carrier is incompatible with the active ingredient, it is contemplated for use in the therapeutic or pharmaceutical composition.

[0056] As used herein, the term "solvate" is intended to include stoichiometric or non-stoichiometric solvent addition forms. If the solvent is water, the solvate formed is a hydrate, and when the solvent is ethanol, the solvate formed is an ethanolate. Hydrates are formed by one or more molecules of water with one molecule of the substance, wherein the water retains its molecular form of HO. Such a combination can form one or more hydrates, such as hemihydrates, monohydrates, and dihydrates.

[0057] As used herein, "prodrug" refers to a chemically modified active or inactive compound that, after administration to a subject, undergoes physiological action in the body (e.g., hydrolysis, necrolysis, etc.) to become a compound of the present invention. The adaptability and technology for making and using prodrugs are well known to those skilled in the art.

[0058] The term "therapeutically effective amount" of the compound of the present invention refers to an amount of the compound of the present invention that can elicit a biological or medical response in an individual or improve symptoms, slow down or delay disease progression, or prevent disease.

[0059] As used herein, the term "subject" refers to an animal. Preferably, the animal is a mammal. In particular, the subject refers to, for example, primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, etc. In a preferred embodiment, the subject is a human.

[0060] As used herein, the term "inhibit" refers to a reduction or suppression of a particular condition, symptom or disorder or disease, or a significant decrease in the baseline activity of a biological activity or process.

[0061] As used herein, in one embodiment, the term "treating" any disease or condition refers to ameliorating the disease or condition (i.e., arresting or slowing the progression of the disease or at least one of its clinical symptoms). In another embodiment, "treating" refers to improving at least one physical parameter, which may not be noticeable to the patient. In another embodiment, "treating" refers to modulating the disease or condition physically (e.g., stabilizing a noticeable symptom) or physiologically (e.g., stabilizing a physical parameter), or both. Beneficial effects

[0062] The main advantages of the present invention are that the compound of the present invention has high selectivity for PARP1, has fewer side effects than olaparib (AZD-2281), and has high clinical application value. DETAILED DESCRIPTION

[0063] Unless otherwise specified, the experimental materials and reagents used in the following examples can be obtained from commercial sources. The starting materials can generally be obtained from commercial sources or easily prepared using methods known to those skilled in the art.

[0064] In each embodiment, the experimental equipment description (such as 1 H NMR was recorded on a Varian Mercury-300 or Varian Mercury-400 nuclear magnetic resonance spectrometer. 13 C NMR was recorded on a Varian Mercury-400, Varian Mercury-500, or Varian Mercury-600 nuclear magnetic resonance instrument, and chemical shifts are expressed in δ (ppm). Mass spectra were recorded on Finnigan / MAT-95 (EI), Finnigan LCQ / DECA, and Micromass Ultra Q-TOF (ESI) mass spectrometers. Reverse-phase preparative HPLC separations used silica gel of 200–300 mesh.

[0065] Abbreviations

[0066] Synthesis of key intermediates

[0067] Intermediate 1a: 7-(Bromomethyl)-8-fluoro-3-methylquinoxalin-2(1H)-one

[0068] Step 1: Synthesis of ethyl (4-bromo-3-fluoro-2-nitrophenyl)alaninate

[0069] To a dry 100 mL single-necked flask, compound 1a-1 (2.38 g, 10 mmol), alanine ethyl ester hydrochloride (1.53 g, 10 mmol), DIEA (3.87 g, 30 mmol), and DMF (50 mL) were added sequentially at room temperature and stirred overnight. The reaction mixture was added with water (100 mL) and extracted with EA (100 mL) three times. The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified on a silica gel column (PE:EA = 4:1) to obtain product 1a-2 (1.5 g, orange solid) in a yield of 45%. 1 H NMR (600MHz, CDCl3): δ7.42-7.38(m,1H),7.19-7.18(m,1H),6.35-6.32(m,1H ),4.19-4.09(m,3H),1.95-1.92(m,1H),1.51-1.48(m,2H),1.24-1.18(m,3H).

[0070] Step 2: Synthesis of 7-bromo-8-fluoro-3-methyl-3,4-dihydroquinoxalin-2(1H)-one

[0071] 1a-2 (3 g, 8.9 mmol), ammonium chloride (3.8 g, 71.6 mmol), methanol (50 mL), and water (5 mL) were added sequentially to a dry 100 mL single-necked flask at room temperature. Zinc powder (4.6 g, 71.6 mmol) was added in portions under ice bath conditions, and the reaction solution was stirred at room temperature for 2 hours. The reaction solution was filtered, and the filtrate was extracted with water (50 mL) and ethyl acetate (50 mL) three times. The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Methanol (15 mL), ethyl acetate (15 mL), and 4M hydrochloric acid (dioxane solution) (1 mL) were added to the residue and stirred at room temperature for 2 hours. The reaction solution was concentrated to give the crude product 1a-3 (2.5 g, brown solid) in a yield of 45%. LCMS (ESI): m / z 257.1 [MH] - ; RT = 1.48 min (3.00 min).

[0072] Step 3: Synthesis of 7-bromo-8-fluoro-3-methylquinoxalin-2(1H)-one

[0073] To a dry 250 mL single-necked flask, 1a-3 (2.5 g, 9.7 mmol) and DCM (160 mL) were added sequentially at room temperature. 2,3-Dichloro-5,6-dicyanobenzoquinone (2.6 g, 11.6 mmol) was added portionwise, and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated, and the residue was dropwise added with saturated sodium carbonate solution (50 mL). The mixture was filtered, and the filter cake was washed with water and dried. The solid was slurried with ethyl acetate and filtered to obtain the product 1a-4 (1.6 g, light brown solid) in a 64% yield. 1 H NMR (600MHz, DMSO-d6): δ12.61 (s 1H), 7.53-7.47 (m, 2H), 2.40 (s, 3H).

[0074] Step 4: Synthesis of 8-fluoro-7-(hydroxymethyl)-3-methylquinoxalin-2(1H)-one

[0075] 1a-4 (1.6 g, 6.25 mmol), (tributyltin)methanol (2.4 g, 7.5 mmol), x-phos Pd G2 (490 mg, 0.625 mmol), and dioxane (40 mL) were added sequentially at room temperature to a dry 100 mL single-necked flask. The mixture was heated at 80°C overnight under nitrogen protection. The reaction solution was concentrated and purified by silica gel column (PE:EA=1:1) to obtain product 1a-5 (1 g, white solid) in a yield of 77%. 1 H NMR (400MHz, DMSO-d6): δ12.42 (s 1H), 7.52-7.50 (m, 1H), 7.34-7.30 (m, 1H), 5.40 (t, J = 5.6Hz, 1H), 4.63 (d, J = 5.2Hz, 2H), 2.41 (s, 3H).

[0076] Step 5: Synthesis of 7-(bromomethyl)-8-fluoro-3-methylquinoxalin-2(1H)-one

[0077] To a dry 250 mL single-necked flask, 1a-5 (800 mg, 3.8 mmol) and dichloromethane (100 mL) were added sequentially at room temperature. Phosphorus tribromide (3.1 g, 11.5 mmol) was then added dropwise in an ice bath. The mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated at low temperature, and the residue was added with water (20 mL). The pH was adjusted to 8 with saturated aqueous sodium carbonate solution, filtered, and the filter cake was washed with water and dried to afford product 1a (600 mg, green solid). Yield: 60%. LCMS (ESI): m / z 269.1 [MH] - ; RT = 1.37 min (3.00 min).

[0078] Intermediate 2a: Synthesis of 7-(bromomethyl)-3-ethyl-8-fluoroquinoxalin-2(1H)-one

[0079] The synthesis method refers to the synthesis of intermediate 1a, except that 2-aminobutyric acid methyl ester hydrochloride is used instead of alanine ethyl ester hydrochloride as the starting material in step 1. LCMS (ESI): m / z 283 [MH] - ; RT = 1.56 min (3.00 min).

[0080] Intermediate 3a: 7-(1-bromoethyl)-8-fluoro-3-methylquinoxalin-2(1H)-one

[0081] Step 1: Synthesis of 7-acetyl-8-fluoro-3-methylquinoxalin-2(1H)-one

[0082] To a single-necked flask containing 10 mL of dioxane, 1a-4 (500 mg, 1.95 mmol), tributyl(1-ethoxyvinyl)stannane (848 mg, 2.34 mmol), and bistriphenylphosphine palladium dichloride (69 mg, 0.10 mmol) were added and reacted at 95°C for 16 hours. After adding dilute hydrochloric acid (2N), the reaction was continued at 50°C for 0.5 hours. After cooling to room temperature, saturated aqueous potassium fluoride solution was added and the reaction was continued for another 0.5 hours. The reaction solution was directly filtered, the filter cake was dried, and then slurried with PE:EA = 1:1 to obtain the product 3a-1 (290 mg, yellow solid). Yield: 67.6%, LCMS (ESI): m / z 221.1 [M+H] + ; RT = 0.792 min (2.50 min).

[0083] Step 2: Synthesis of 8-fluoro-7-(1-hydroxyethyl)-3-methylquinoxalin-2(1H)-one

[0084] 3a-1 (0.15 g, 0.68 mmol) was added to a 100 mL single-necked flask containing 10 mL of methanol. Sodium borohydride (39 mg, 1.02 mmol) was slowly added under ice-cooling and allowed to react at room temperature for 1 hour. The reaction mixture was quenched with saturated ammonium chloride solution and extracted three times with dichloromethane. The organic phase was washed twice with water and once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 1:1) to afford the crude product 3a-2 (100 mg, yellow solid). Yield: 66.1%, LCMS (ESI): m / z 223.1 [M+H] + ; RT = 1.12 min (2.50 min).

[0085] Step 3: Synthesis of 7-(1-bromoethyl)-8-fluoro-3-methylquinoxalin-2(1H)-one

[0086] To a 100 mL single-necked flask containing 6 mL of dichloromethane was added 3a-2 (100 mg, 0.45 mmol), triphenylphosphine (354 mg, 1.35 mmol), and carbon tetrabromide (299 mg, 0.90 mmol). The mixture was allowed to react at room temperature for 2 hours. The reaction mixture was directly concentrated and subjected to column chromatography (PE:EA = 2:1) to afford product 3a (70 mg, yellow solid). Yield: 54.7%, purity: 50%. LCMS (ESI): m / z 283.1 [MH] - ; RT = 1.234 min (2.50 min).

[0087] Intermediate 4a: 7-(1-bromoethyl)-8-fluoro-3-ethylquinoxalin-2(1H)-one

[0088] The synthesis method refers to the synthesis of intermediate 3a, except that replace As the starting material, 1a'-4 is obtained by using 2-aminobutyric acid methyl ester hydrochloride instead of alanine ethyl ester hydrochloride as the starting material in step 1 of the preparation of intermediate 1a. LCMS (ESI): m / z 296.9 [MH] - ; RT = 1.574 (3.00 min).

[0089] Intermediate 5a: Methyl (3-ethyl-8-fluoro-2-oxo-1,2-dihydro-1,6-naphthyridin-7-yl)methanesulfonate

[0090] Step 1: Synthesis of (4-amino-6-chloro-5-fluoropyridin-3-yl)methanol

[0091] Under nitrogen, 5a-1 (5.0 g, 22.9 mmol) and anhydrous toluene (70 mL) were added to a 500 mL three-necked flask at room temperature. Diisopropylaluminum hydride (91.5 mmol, 91.5 mL) was slowly added dropwise at -78°C and allowed to react for 16 hours. Methanol (3.66 mL), 15% sodium hydroxide solution (3.66 mL), and methanol (3.66 mL) were added dropwise in sequence under an ice bath. The reaction solution was concentrated and purified by column chromatography (PE:EA = 1:1) to afford 5a-2 (2.45 g, white solid) in a 61.2% yield. LCMS (ESI): m / z 177.1 [M+H] + ; RT = 0.707 min (2.50 min).

[0092] Step 2: Synthesis of 4-amino-6-chloro-5-fluoronicotinaldehyde

[0093] Compound 5a-2 (2.45 g, 13.9 mmol) and manganese dioxide (12 g, 139 mmol) were added to a single-necked flask containing 40 mL of DCM at room temperature. The mixture was stirred at room temperature and allowed to react for 16 hours. The reaction solution was filtered and concentrated to afford compound 5a-3 (2.2 g, gray solid) in a 90.8% yield. LCMS (ESI): m / z 175.1 [M+H] + ; RT = 1.210 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ9.95 (d, J = 1.6Hz, 1H), 8.37 (s, 1H), 7.97 (s, 2H).

[0094] Step 3: Synthesis of 7-chloro-3-ethyl-8-fluoro-1,6-naphthyridin-2(1H)-one

[0095] In a dry 250 mL three-necked flask, compound 5a-3 (3.0 g, 17.2 mmol), 4-dimethylaminopyridine (207 mg, 1.7 mmol), and DIEA (15.5 g, 15.5 mmol) were added under an ice bath. Butyryl chloride (12.8 g, 120 mmol) was slowly added dropwise. The mixture was stirred at room temperature and allowed to react overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (PE:EA = 10:1) to obtain compound 5a-4 (1.3 g, yellow solid) in a 33.4% yield. 1 H NMR (400MHz, DMSO-d6): δ12.47(s,1H),8.52(s,1H),7.89(s,1H),2.54-2.49(m,2H),1.19-1.16(m,3H).

[0096] Step 4: Synthesis of methyl 3-ethyl-8-fluoro-2-oxo-1,2-dihydro-1,6-naphthyridine-7-carboxylate

[0097] To a dry 50 mL single-necked flask, 5a-4 (370 mg, 1.63 mmol), methanol (10 mL), triethylamine (0.68 mL, 4.90 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (237 mg, 0.33 mmol) were added sequentially at room temperature. Carbon monoxide was replaced three times, and the reaction was continued at 80°C for 24 hours. The reaction solution was concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA = 2:1) to obtain 5a-5 (190 mg, yellow solid) in a yield of 46.51%. LCMS (ESI): m / z 251.0 [M+H]. + ; RT = 1.114 min (2.50 min). 1H NMR (400MHz, DMSO-d6): δ12.52(s,1H),8.72(s,1H),7.92(s,1H),3.91(s,3H),2.58-2.54(m,2H),1.19(t,J=7.6Hz,3H).

[0098] Step 5: Synthesis of 3-ethyl-8-fluoro-7-(hydroxymethyl)-1,6-naphthyridin-2(1H)-one

[0099] In a dry 25 mL single-necked flask, 5a-5 (190 mg, 0.76 mmol), tetrahydrofuran (5 mL), and lithium borohydride-tetrahydrofuran (2.0 M, 0.76 mmol, 1.52 mmol) were added sequentially at 0°C. The mixture was stirred at room temperature and reacted for 2 hours. The mixture was quenched with saturated water (20 mL) and extracted with EA (30 mL × 2). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative plate (pure ethyl acetate) to give 5a-6 (60 mg, nearly white solid) in a yield of 35.56%. LCMS (ESI): m / z 223.1 [M+H] + ; RT = 0.875 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ12.23(s,1H),8.61(s,1H),7.87(s,1H),5.34(t,J=6.0Hz,1H),4.63-4.61(m,2H),2.55-2.51(m,2H),1.18(t,J=7.6Hz,3H).

[0100] Step 6: Synthesis of methyl (3-ethyl-8-fluoro-2-oxo-1,2-dihydro-1,6-naphthyridin-7-yl)methanesulfonate

[0101] 5a-6 (40 mg, 0.18 mmol), tetrahydrofuran (3 mL), triethylamine (0.08 mL, 0.54 mmol) and methanesulfonyl chloride (0.02 mL, 0.22 mmol) were added sequentially to a dry 25 mL single-necked flask at room temperature. The mixture was stirred at room temperature and reacted for 2 hours. The mixture was diluted with water (10 mL) and extracted with EA (10 mL × 2). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 5a (48 mg, yellow solid). LCMS (ESI): m / z 301.1 [M+H] + ; RT = 1.170 min (2.50 min).

[0102] Intermediate 6a: (R)-6-Fluoro-N-methyl-5-(2-methylpiperazin-1-yl)picolinamide hydrochloride

[0103] Step 1: Synthesis of methyl 5-bromo-6-fluoropicolinate

[0104] To a 50 mL single-necked flask, compound 6a-1 (1 g, 4.6 mmol), acetonitrile (30 mL), and silver difluoride (1.76 g, 13.9 mmol) were added sequentially and stirred overnight. The reaction mixture was filtered, the filtrate concentrated, and purified on a silica gel column (PE:EA = 5:1) to afford 6a-2 (450 mg, white solid) in a 42% yield. LCMS (ESI): m / z 233.9 [M+H] + ; RT = 1.51 min (3.00 min).

[0105] Step 2: Synthesis of tert-butyl (R)-4-(2-fluoro-6-(methoxycarbonyl)pyridin-3-yl)-3-methylpiperazine-1-carboxylate

[0106] To a 50 mL single-necked flask were added 6a-2 (450 mg, 1.9 mmol), (R)-4-Boc-2-methylpiperazine (577 mg, 2.9 mmol), Ruphos Pd G3 (159 mg, 0.19 mmol), cesium carbonate (1.5 g, 4.7 mmol), and dioxane (6 mL). The mixture was heated at 80°C overnight under nitrogen. The reaction mixture was concentrated and purified on a silica gel column (PE:EA = 2:1) to afford product 6a-3 (300 mg, yellow solid) in a 44% yield. LCMS (ESI): m / z 354.1 [M+H]. + ; RT = 1.80 min (3.00 min).

[0107] Step 3: Synthesis of tert-butyl (R)-4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)-3-methylpiperazine-1-carboxylate

[0108] Compound 6a-3 (100 mg, 0.28 mmol) and a methylamine ethanol solution (2 mL) were added sequentially to a 50 mL single-necked flask, and the reaction mixture was stirred overnight. The reaction mixture was concentrated to afford 6a-4 (80 mg, yellow solid) in an 80% yield. LCMS (ESI): m / z 297.1 [M-56+H] + ; RT = 1.70 min (3.00 min).

[0109] Step 4: Synthesis of (R)-6-fluoro-N-methyl-5-(2-methylpiperazin-1-yl)picolinamide hydrochloride

[0110] To a 20 mL single-necked flask, compound 6a-4 (80 mg, 0.23 mmol), EA (2 mL), and 4 M hydrochloric acid (in dioxane) (2 mL) were added sequentially and stirred at room temperature for 2 hours. The reaction mixture was concentrated to afford 6a (65 mg, yellow oil) in a 100% yield. LCMS (ESI): m / z 253.2 [M+H] + ; RT = 0.99 min (3.00 min).

[0111] Intermediate 7a: Synthesis of (R)-N-methyl-5-(2-methylpiperazin-1-yl)pyridineamide hydrochloride

[0112] The synthesis method refers to the synthesis of intermediate 6a, except that step 1 was not performed and 6a-1 was directly used instead of 6a-2 in step 2 and the synthesis was started from step 2. LCMS (ESI): m / z 235.1 [M+H] + ; RT = 0.82 min (3.00 min). 1 H NMR (400MHz, DMSO-d6): δ12.42 (s 1H), 7.52-7.50 (m, 1H), 7.34-7.30 (m, 1H), 5.40 (t, J = 5.6Hz, 1H), 4.63 (d, J = 5.2Hz, 2H), 2.41 (s, 3H).

[0113] Intermediate 8a: Synthesis of N-methyl-5-(piperazin-1-yl)pyridineamide hydrochloride

[0114] The synthesis method refers to the synthesis of intermediate 6a, except that step 1 is omitted and 6a-1 is directly used instead of 6a-2 in step 2. 4-Boc-piperazine is used instead of (R)-4-Boc-2-methylpiperazine and the synthesis is started from step 2 (please confirm whether it is correct). LCMS (ESI): m / z 221.2 [M+H] + ; RT = 0.285 min (6.00 min).

[0115] Intermediate 9a: (S)-N-methyl-5-(2-methylpiperazin-1-yl)picolinamide hydrochloride

[0116] The synthesis method refers to the synthesis of intermediate 6a, except that step 1 is omitted and 6a-1 is directly used instead of 6a-2 in step 2. (S)-4-Boc-2-methylpiperazine is used instead of (R)-4-Boc-2-methylpiperazine and the synthesis is started from step 2. LCMS (ESI): m / z 235.2 [M+H] +; RT = 0.307 min (3.00 min).

[0117] Intermediate 10a: (S)-N-methyl-5-(3-methylpiperazin-1-yl)methylpicolinamide hydrochloride

[0118] The synthesis method refers to the synthesis of intermediate 6a, except that step 1 was omitted and 6a-1 was directly used instead of 6a-2 in step 2. Tert-butyl-(S)-2-methylpiperazine-1-carboxylate was used instead of (R)-4-Boc-2-methylpiperazine and the synthesis was started from step 2. LCMS (ESI): m / z 235.2 [M+H] + ; RT = 0.81 min (3.00 min).

[0119] Intermediate 11a: (R)-N-methyl-5-(3-methylpiperazin-1-yl)methylpicolinamide hydrochloride

[0120] The synthesis method refers to the synthesis of intermediate 6a, except that step 1 is not performed, and 6a-1 is directly used instead of 6a-2 in step 2, and tert-butyl-(R)-2-methylpiperazine-1-carboxylate is used instead of (R)-4-Boc-2-methylpiperazine to start the synthesis from step 2.

[0121] Intermediate 12a: 6-Fluoro-N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride

[0122] Step 1: Synthesis of tert-butyl 4-(2-fluoro-6-(methoxycarbonyl)pyridin-3-yl)piperazine-1-carboxylate

[0123] To a dry 50 mL single-necked flask were added 6a-2 (500 mg, 2.1 mmol), 1-tert-butyloxycarbonylpiperazine (600 mg, 3.2 mmol), Ruphos Pd G3 (180 mg, 0.21 mmol), cesium carbonate (1.7 g, 5.2 mmol), and dioxane (15 mL). The mixture was heated at 80°C overnight under nitrogen. The reaction mixture was concentrated and purified on a silica gel column (PE:EA = 2:1) to afford 12a-1 (640 mg, yellow solid) in an 88% yield. LCMS (ESI): m / z 340.1 [M+H]. + ; RT = 1.74 min (3.00 min).

[0124] Step 2: Synthesis of 5-(4-(tert-Butyloxycarbonyl)piperazin-1-yl)-6-fluoropicolinic acid

[0125] To a dry 50 mL single-necked flask, 12a-1 (320 mg, 0.94 mmol) and tetrahydrofuran (8 mL) were added sequentially, followed by the dropwise addition of an aqueous solution (8 mL) of lithium hydroxide monohydrate (200 mg, 4.7 mmol). The reaction mixture was stirred for 2 hours. The pH was adjusted to 6 with 1 M hydrochloric acid solution, and the reaction mixture was concentrated and passed through a reverse phase column (2% to 40% acetonitrile in water) to afford 12a-2 (300 mg, yellow solid) in a 92% yield. LCMS (ESI): m / z 326.1 [M+H] + ; RT = 1.26 min (3.00 min).

[0126] Step 3: Synthesis of tert-butyl 4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazine-1-carboxylate

[0127] To a dry 50 mL single-necked flask, compound 12a-2 (300 mg, 0.92 mmol), 1-hydroxybenzotriazole (149 mg, 1.1 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (211 mg, 1.1 mmol), DIEA (237 mg, 1.84 mmol), DMF (6 mL), and methylamine hydrochloride (123 mg, 1.84 mmol) were added sequentially and stirred at room temperature overnight. Water (20 mL) was added to the reaction solution, and EA was extracted (30 mL x 3). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and passed through a reverse phase column (20% to 70% acetonitrile in water) to afford compound 12a-3 (250 mg, white solid) in 80% yield. LCMS (ESI): m / z 283.1 [M-100+H]. + ; RT = 1.66 min (3.00 min).

[0128] Step 4: Synthesis of 6-fluoro-N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride

[0129] Compound 12a-3 (250 mg, 0.74 mmol), ethyl acetate (3 mL), and 4M hydrochloric acid (dioxane solution) (1 mL) were added sequentially to a dry 20 mL single-necked flask. The mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated to afford 12a (200 mg, yellow solid) in a 100% yield. LCMS (ESI): m / z 239.1 [M+H] + ; RT = 0.91 min (3.00 min). 1H NMR (600MHz, CD3OD): δ7.98-7.96(m,1H),7.67-7.64(m,1H),3.50-3.48(m,4H),3.45-3.43(m,4H),2.93(d,J=3.6Hz,3H).

[0130] Intermediate 13a: Methyl (R)-6-fluoro-5-(3-methylpiperazin-1-yl)picolinate hydrochloride

[0131] The synthesis method was similar to that of intermediate 12a, except that tert-butyl (R)-2-methylpiperazine-1-carboxylate was used instead of 1-tert-butyloxycarbonylpiperazine. LCMS (ESI): m / z 253.1 [M+H] + ;RT=0.342&0.576min(6.00min).

[0132] 1 H NMR (400MHz, DMSO-d6): δ9.62(s,1H),9.47(s,1H),8.45(dd,J=4.4,9.2Hz,1H),7.87(dd,J=1.2,8.0Hz,1H),7.72-7.67(m,1 H),3.63-3.59(m,2H),3.39-3.45(m,2H),3.24-3.13(m,2H),3.04-2.98(m,1H),2.78(d,J=4.8Hz,3H),1.31(d,J=6.4Hz,3H).

[0133] Intermediate 14a: 7-(1-bromoethyl)-3-methylquinoxalin-2(1H)-one

[0134] The synthesis method refers to the synthesis of intermediate 3a, except that replace As the starting material, 14a-4 was obtained by using 4-bromo-1-fluoro-2-nitrobenzene instead of 1a-1 as the starting material in step 1 of the preparation of intermediate 1a. LCMS (ESI): m / z 267.0 [M+H] + ; RT = 1.327 min (2.50 min).

[0135] Intermediate 15a: 7-(1-bromoethyl)-3-ethylquinoxalin-2(1H)-one

[0136] The synthesis method refers to the synthesis of intermediate 3a, except that replace As the starting material, 15a-4 was obtained by replacing 1a-1 with 4-bromo-1-fluoro-2-nitrobenzene and replacing ethyl alanine hydrochloride with methyl 2-aminobutyrate hydrochloride in step 1 of the preparation of intermediate 1a. LCMS (ESI): m / z 296.9 [MH] - ; RT = 1.574 min (2.50 min).

[0137] Intermediate 16a: Synthesis of 6-chloro-N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride

[0138] Step 1: Synthesis of 5-bromo-6-chloro-N-methylpicolinamide

[0139] 16a-1 (1.00 g, 3.99 mmol) and a methylamine ethanol solution (33 wt %, 10 mL) were added to a flask in sequence. The reaction was allowed to proceed at room temperature for 16 hours. The reaction solution was concentrated to afford 16a-2 (0.94 g, yellow oil) in a 94.37% yield. LCMS (ESI): m / z 250.9 [M+H] + ; RT = 1.448 min (2.50 min).

[0140] Step 2: Synthesis of tert-butyl 4-(2-chloro-6-(methylcarbamoyl)pyridin-3-yl)piperazine-1-carboxylate

[0141] To a flask were added 16a-2 (620 mg, 2.49 mmol), toluene (15 mL), tert-butyl piperazine-1-carboxylate (370 mg, 1.99 mmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (155 mg, 0.25 mmol), cesium carbonate (2020 mg, 6.21 mmol), and palladium acetate (56 mg, 0.25 mmol). The mixture was reacted at 100°C under nitrogen for 16 hours. The reaction mixture was filtered and the filtrate was concentrated. Purification by silica gel column chromatography (PE:EA = 1:1) afforded 16a-3 (180 mg, yellow oil) in a 20.41% yield. LCMS (ESI): m / z 355.1 [M+H] + ; RT = 1.679 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ8.46(d,J=4.8Hz,1H),7.95(d,J=8.4Hz,1H),7.68(d,J =8.4Hz,1H),3.50(s,4H),3.06-3.04(m,4H),2.80(d,J=4.8Hz,3H),1.43(s,9H).

[0142] Step 3: Synthesis of 6-chloro-N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride

[0143] 16a-3 (180 mg, 0.51 mmol), DCM (2 mL) and dioxane hydrochloride (4.0 M, 2 mL) were added to a dry flask. The mixture was stirred at room temperature for 2 hours and then concentrated to afford 16a (140 mg, yellow solid). Yield: 94.59%, LCMS (ESI): m / z 255.1 [M+H] + ;RT=0.336min&0.461min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ9.40 (s, 2H), 8.50 (d, J = 4.4Hz, 1H), 7.97 (d, J = 8.4Hz, 1 H),7.76(d,J=8.4Hz,1H),3.34-3.32(m,4H),3.25(m,4H),2.80(d,J=4.8Hz,3H).

[0144] Intermediate 17a: Synthesis of N,6-dimethyl-5-(piperazin-1-yl)picolinamide hydrochloride

[0145] Step 1: Synthesis of 5-bromo-6-methylpicolinic acid

[0146] To a dry flask were added 17a-1 (500 mg, 2.54 mmol), methanol (6 mL), water (3 mL), and sodium hydroxide (507 mg, 12.69 mmol) in sequence. The mixture was reacted at 70°C for 1 hour. The reaction solution was concentrated under reduced pressure. The mixture was diluted with water (10 mL) and adjusted to pH 4 with 3M dilute hydrochloric acid. The filter cake was filtered to obtain 17a-2 (300 mg, white solid). Yield: 54.72%. LCMS (ESI): m / z 218.0 [M+H] + ; RT = 1.208 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ8.12(d,J=8.0Hz,1H),7.78(d,J=8.4Hz,1H),2.67(s,3H).

[0147] Step 2: Synthesis of 5-bromo-N,6-dimethylpicolinamide

[0148] To a dry flask were added 17a-2 (300 mg, 1.39 mmol), DMF (3 mL), DIEA (0.92 mL, 5.55 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (792 mg, 2.08 mmol), and a 2.0 M solution of methylamine in tetrahydrofuran (1.39 mL, 2.78 mmol). The reaction was allowed to react at room temperature for 1 hour. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (10 mL x 2). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified using a preparative plate (PE:EA = 3:2) to afford 17a-3 (215 mg, yellow solid) in a 67.59% yield. LCMS (ESI): m / z 231.0 [M+H]. + ; RT = 1.427 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ8.67(d,J=4.0Hz,1H), 8.18(d,J=8.4Hz,1H), 7.75(d,J=8.4Hz,1H), 2.82(d,J=4.8Hz,3H), 2.65(s,3H).

[0149] Step 3: Synthesis of tert-butyl 4-(2-methyl-6-(methylcarbamoyl)pyridin-3-yl)piperazine-1-carboxylate

[0150] To a dry flask were added 17a-3 (200 mg, 0.87 mmol), toluene (8 mL), tert-butyl piperazine-1-carboxylate (179 mg, 0.96 mmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (54 mg, 0.09 mmol), cesium carbonate (711 mg, 2.18 mmol), and palladium acetate (20 mg, 0.09 mmol). The mixture was reacted at 100°C under nitrogen for 16 hours. The reaction mixture was filtered and the filtrate was concentrated. The residue was purified using a preparative plate (PE:EA = 1:1) to afford 17a-4 (160 mg, yellow solid) in a 54.80% yield. LCMS (ESI): m / z 335.1 [M+H]. + .

[0151] Step 4: Synthesis of N,6-dimethyl-5-(piperazin-1-yl)picolinamide hydrochloride

[0152] 17a-4 (160 mg, 0.48 mmol), DCM (2 mL), and hydrochloric acid-dioxane (4.0 M, 2 mL) were added to a flask. The mixture was reacted at room temperature for 4 hours and then concentrated to afford 17a (129 mg, yellow solid). Yield: 99.58%, LCMS (ESI): m / z 235.2 [M+H]+ ;RT=0.340min&0.450min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ9.44(s,2H),8.56(d,J=4.8Hz,1H),7.88(d,J=8.0Hz,1 H),7.61(d,J=8.4Hz,1H),3.25-3.16(m,8H),2.82(d,J=4.4Hz,3H),2.54(s,3H).

[0153] Synthesis of compound 2

[0154] To a dry 25 mL single-necked flask, compound 2a (50 mg, 0.175 mmol), 7a (60 mg, 0.22 mmol), DIEA (68 mg, 0.525 mmol), and acetonitrile (3 mL) were added sequentially at room temperature. The mixture was heated at 80°C for 2 hours. The reaction solution was purified using a reverse-phase preparative column and dried to afford compound 2 (25 mg, 0.056 mmol) as a white solid in a 39% yield. LCMS (ESI): m / z 439.1 [M+H] + ; RT = 1.62 min (3.00 min). 1 H NMR (600MHz, DMSO-d6): δ12.45(s,1H),8.36(d,J=3.2Hz,1H),8.19(s,1H),7.81(d,J=5.6Hz ,1H),7.57(d,J=5.2Hz,1H),7.31-7.35(m,2H),4.22(s,1H),3.68(s,2H),3.58(d,J=8.0Hz,1 H),3.04-3.07(m,1H),2.92-2.94(m,1H),2.80-2.84(m,2H),2.78(d,J=3.2Hz,3H),2.75(d,J =7.6Hz,1H),2.36-2.37(m,1H),2.20-2.23(m,1H),1.21-1.23(m,3H),1.10(d,J=4.4Hz,3H).

[0155] Synthesis of Compound 5-Compound 10

[0156] According to the method described in compound 2, intermediates 2a and 7a were replaced with the intermediates in the following table to synthesize compounds 5-10. The structural formulas of the compounds in the examples are shown in the previous table.

[0157] Synthesis of compound 12

[0158] To a dry 25 mL single-necked flask, 5a (27 mg, 0.09 mmol), acetonitrile (3 mL), 8a (35 mg, 0.13 mmol), and DIEA (0.07 mL, 0.45 mmol) were added sequentially at room temperature. The mixture was heated to 75°C for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by prep-HPLC (0.1% formic acid) to afford 12 (11.06 mg, white solid) in a 28.98% yield. LCMS (ESI): m / z 425.05 [M+H] + ; RT = 3.134 min (6.00 min). 1 H NMR (400MHz, DMSO-d6): δ12.25(s,1H),8.62(s,1H),8.40-8.37(m,1H),8.24(d,J=2.4Hz,1H),7.87(s,1H),7.81(d,J=8.8Hz,1H),7.37-7 .34(m,1H),3.76(d,J=2.4Hz,2H),3.30-3.26(m,4H),2.77(d,J=4.8Hz,3H),2.63-2.61(m,4H),2.55-2.51(m,2H),1.18(t,J=7.6Hz,3H).

[0159] Synthesis of compounds 13B1 and 13B2

[0160] Compound 13 (100 mg) was separated by chiral column, column model 100*3.0mm 3.0μm, mobile phase: methanol (+0.1% diethylamine), to obtain compound 13B1 (32.23mg) and compound 13B2 (38mg);

[0161] Compound 13B1: peak time 4.125 min; ee%>99, LCMS (ESI): m / z 457.30 [M+H] + ;(DMSO-d6): δ8.39-8.38(m,1H),7.83(d,J=7.6Hz,1H),7.57-7.52(m,2H),7.31-7.27(m,1H),3.98-3.96(m,1H),3 .15(s,4H),2.84-2.79(m,2H),2.76(d,J=4.8Hz,3H),2.62-2.51(m,4H),1.40(d,J=6.8Hz,3H),1.24-1.20(m,3H);

[0162] Compound 13B2: peak time 5.635 min; ee%>99, LCMS (ESI): m / z 457.20 [M+H] + ;(DMSO-d6): δ12.46(s,1H),8.40-8.39(m,1H),7.83(d,J=8.0Hz,1H),7.60-7.53(m,2H),7.35(s,1H),3.99(s, 1H),3.16(s,4H),2.89-2.82(m,2H),2.76(d,J=4.8Hz,3H),2.68-2.52(m,4H),1.38(s,3H),1.24-1.17(m,3H).

[0163] Synthesis of compound 15

[0164] According to the method described in compound 12, intermediate 8a was replaced by 12a to synthesize compound 15

[0165] Synthesis of compound 23

[0166] Step 1: Synthesis of 7-acetyl-3-ethyl-8-fluoro-1,6-naphthyridin-2(1H)-one

[0167] Compound 5a-4 (500 mg, 2.2 mmol), tributyltin (1.6 g, 4.4 mmol), and bistriphenylphosphine palladium dichloride (240 mg, 0.34 mmol) were added to a 100 mL single-necked bottle containing 10 mL of dioxane. The mixture was reacted at 95°C for 16 hours. The reaction solution was filtered, concentrated, and purified by column chromatography (PE:EA = 1:1) to afford compound 23-1 (490 mg, yellow solid). Yield: 94.7%, LCMS (ESI): m / z 235.1 [M+H] + ; RT = 1.050 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ12.40(s,1H),8.67(s,1H),7.87(s,1H),2.56(s,3H),2.51-2.48(m,2H),1.13-1.09(m,3H).

[0168] Step 2: Synthesis of 3-ethyl-8-fluoro-7-(1-hydroxyethyl)-1,6-naphthyridin-2(1H)-one

[0169] In a dry 100 mL single-necked flask, 23-1 (40 mg, 0.17 mmol), methanol (3 mL), and sodium borohydride (7.8 mg, 0.20 mmol) were added sequentially under ice bath. The mixture was stirred at room temperature for 5 hours. The mixture was quenched with saturated ammonium chloride (1 mL), the pH was adjusted to neutral, the mixture was diluted with water, and the mixture was extracted with ethyl acetate (10 mL x 5). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 23-2 (40 mg, gray solid). Yield: 100%, LCMS (ESI): m / z 237.1 [M+H] + ; RT = 0.820 min (2.50 min).

[0170] Step 3: Synthesis of 7-(1-bromoethyl)-3-ethyl-8-fluoro-1,6-naphthyridin-2(1H)-one

[0171] To a 100 mL single-necked flask containing 5 mL of dichloromethane was added 23-2 (100 mg, 0.42 mmol), triphenylphosphine (341 mg, 1.3 mmol), and carbon tetrabromide (279 mg, 0.84 mmol). The mixture was allowed to react at room temperature for 2 hours. The reaction mixture was then concentrated to afford 23-3 (100 mg, gray solid). Yield: 79%, LCMS (ESI): m / z 297.0 [MH] - ; RT = 1.521 min (2.50 min).

[0172] Step 4: Synthesis of 5-(4-(1-(3-ethyl-8-fluoro-2-oxo-1,2-dihydro-1,6-naphthyridin-7-yl)ethyl)piperazin-1-yl)-6-fluoro-N-methylpicolinamide

[0173] In a 25 mL single-necked bottle containing 5 mL of acetonitrile, 23-3 (100 mg, 0.33 mmol), 12a (91 mg, 0.33 mmol) and N,N-diisopropylethylamine (216 mg, 1.67 mmol) were added. The reaction was carried out at 70°C for 16 hours. The reaction solution was concentrated and analyzed by Pre-HPLC (formic acid) to give compound 23 (2.17 mg, yellow solid). Yield: 1.4%. LCMS (ESI): m / z 457.35 [M+H] + ; RT = 3.257 min (6.00 min). 1H NMR (400MHz, DMSO-d6): δ8.66(s,1H),8.38(d,J=4.4Hz,1H),7.87(s,1H),7.82(d,J=8.0Hz,1H),7.54-7.49(m,1H),4.25(d,J=6.0 Hz,1H),3.11(d,J=3.2Hz,4H),2.76(d,J=4.8Hz,3H),2.62(s,4H),2.55(d,J=7.2Hz,2H),1.45(d,J=6.8Hz,3H),1.20-1.17(m,3H).

[0174] Synthesis of compounds 23B1 and 23B2

[0175] Compound 23 (500 mg) was separated by chiral column, column model 100*3.0mm 3.0μm, mobile phase: ethanol (+0.1% diethylamine), to obtain compound 23B1 (173mg) and compound 23B2 (168mg);

[0176] Compound 23B1: peak time 2.766 min; ee%>99%, LCMS (ESI): m / z 457.80 [M+H] + ;(DMSO-d6): δ12.23(s,1H),8.65(s,1H),8.37(d,J=4.8Hz,1H),7.96(s,1H),7.81(d,J=8.0Hz,1H),7.53-7.48(m,1H),4.25-4.2 3(m,1H),3.11(d,J=3.6Hz,4H),2.75(d,J=4.8Hz,3H),2.61(s,4H),2.55-2.54(m,2H),1.45(d,J=6.8Hz,3H),1.20-1.16(m,3H).

[0177] Compound 23B2: peak time 3.723 min; ee%>99%, LCMS (ESI): m / z 457.40 [M+H] +; (DMSO-d6): δ12.23(s,1H),8.66(s,1H),8.37(d,J=4.8Hz,1H),7.86(s,1H),7.81(d,J=8.0Hz,1H),7.53-7.48(m,1H),4.25-4.2 3(m,1H),3.11(d,J=3.2Hz,4H),2.75(d,J=4.8Hz,3H),2.61(s,4H),2.57-2.55(m,2H),1.45(d,J=6.8Hz,3H),1.20-1.16(m,3H).

[0178] Synthesis of compound 29

[0179] Step 1: Synthesis of 7-chloro-8-fluoro-3-methyl-1,6-naphthyridin-2(1H)-one

[0180] In a dry 250 mL three-necked flask, 5a-3 (500 mg, 2.87 mmol), 4-dimethylaminopyridine (35 mg, 0.29 mmol), and N,N-diisopropylethylamine (2.2 g, 17.24 mmol) were added under ice-cooling. Propionyl chloride (1.6 g, 17.24 mmol) was slowly added dropwise and stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by slurrying (PE:EA = 10:1) to obtain 29-1 (300 mg, yellow solid) in a yield of 49.1%. LCMS (ESI): m / z 213.0 [M+H] + ; RT = 1.183 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ8.47(s,1H),7.91(s,1H),2.11(d,J=1.2Hz,3H).

[0181] Step 2: Synthesis of 8-fluoro-3-methyl-2-oxo-1,2-dihydro-1,6-naphthyridine-7-carboxylic acid methyl ester

[0182] In a dry 50 mL single-necked flask, 29-1 (360 mg, 1.7 mmol), methanol (25 mL), triethylamine (0.71 mL, 5.1 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride (370 mg, 0.51 mmol) were added in sequence at room temperature. Carbon monoxide (2 MPa) was replaced three times, and the mixture was heated to 80°C for 24 hours. The reaction solution was concentrated under reduced pressure. The residue was purified by chromatography column (PE:EA=1:2) to obtain 29-2 (200 mg, yellow solid) with a yield of 49.9%. 1H NMR (400MHz, DMSO-d6): δ12.55(s,1H),8.68(s,1H),7.99(s,1H),3.91(s,3H),2.15(s,3H).

[0183] Step 3: Synthesis of 8-fluoro-7-(hydroxymethyl)-3-methyl-1,6-naphthyridin-2(1H)-one

[0184] In a dry 25 mL three-necked flask at 0°C, 29-2 (200 mg, 0.85 mmol), tetrahydrofuran (10 mL) and lithium borohydride-tetrahydrofuran (2.0 M, 1.69 mmol, 0.85 mL) were added in sequence. The reaction was allowed to proceed at room temperature for 6 hours under nitrogen protection. The mixture was quenched with saturated ammonium chloride (0.5 mL) and extracted with EA (30 mL × 2). The organic phase was collected, washed with saturated brine, dried, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative plate (EA) to give compound 29-3 (180 mg, gray solid), yield: 100%, LCMS (ESI): m / z 209.1 [M+H] + ; RT = 0.790 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ12.27(s,1H),8.57(s,1H),7.91(s,1H),5.35-5.34(m,1H),4.62-4.60(m,2H),2.12(s,3H).

[0185] Step 4: Synthesis of (8-fluoro-3-methyl-2-oxo-1,2-dihydro-1,6-naphthyridin-7-yl)methyl methanesulfonate

[0186] In a dry 25 mL three-necked flask, 29-3 (80 mg, 0.38 mmol), tetrahydrofuran (3 mL), triethylamine (0.13 mL, 0.95 mmol), and methanesulfonyl chloride (53 mg, 0.46 mmol) were added sequentially at room temperature. The mixture was stirred at room temperature and reacted for 3 hours. The mixture was diluted with water (10 mL) and extracted with EA (10 mL × 2). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the crude product 29-4 (50 mg, yellow solid). LCMS (ESI): m / z 285.1 [MH] - ; RT = 1.079 min (2.50 min).

[0187] Step 5: Synthesis of 6-fluoro-5-(4-((8-fluoro-3-methyl-2-oxo-1,2-dihydro-1,6-naphthyridin-7-yl)methyl)piperazin-1-yl)-N-methylpicolinamide

[0188] To a dry 25 mL single-necked flask was added compound 29-4 (50 mg, 0.17 mmol), acetonitrile (4 mL), compound 12a (40 mg, 0.17 mmol), DIEA (88 mg, 0.68 mmol), and sodium iodide (102 mg, 0.68 mmol) in sequence at room temperature. The mixture was heated to 70°C and allowed to react for 2 hours. The reaction solution was concentrated under reduced pressure. The residue was purified by prep-HPLC (0.1% FA) to afford compound 29 (2 mg, white solid) in a 2.7% yield. LCMS (ESI): m / z 429.20 [M+H] + ; RT = 3.152 min (6.00 min). 1 H NMR (400MHz, DMSO-d6): δ12.37(s,1H),8.63(s,1H),8.43(d,J=4.8Hz,1H),7.94(s,1H),7.85(d,J=8.0Hz ,1H),7.60-7.55(m,1H),3.83-3.77(m,2H),3.21(s,4H),2.77(d,J=4.4Hz,3H),2.51(s,4H),2.14(s,3H).

[0189] Synthesis of compound 30

[0190] The same synthesis method as compound 23 was used, except that the starting material was 29-1 instead of 5a-4. LCMS (ESI): m / z 443.20 [M+H] + ; RT = 3.175 min (6.00 min). 1 H NMR (400MHz, DMSO-d6): δ8.61(s,1H),8.50(s,1H),8.38-8.36(m,1H),7.90(s,1H),7.81(d,J=7.6Hz,1H),7.53-7.48(m ,1H),4.24-4.23(m,1H),3.11(d,J=3.6Hz,4H),2.75(d,J=4.8Hz,3H),2.67(s,4H),2.12(s,3H),1.44(d,J=6.4Hz,3H).

[0191] Synthesis of compounds 36 and 37

[0192] According to the method described in compound 12, intermediates 5a and 8a were replaced with the intermediates in the following table to synthesize compounds 36-37. The structural formulas of the compounds in the examples are shown in the previous table.

[0193] Synthesis of compound 38

[0194] Compound 23-1 (60 mg, 0.29 mmol) and 17a (90 mg, 0.38 mmol) were added to a single-necked flask containing 5 mL of dichloromethane. After reacting for half an hour, the mixture was concentrated and tetraisopropyl titanate (3 mL) was added. The reaction was continued at 80°C for 4 hours. After cooling to room temperature, ethanol (5 mL) and sodium cyanoborohydride (32 mg, 0.52 mmol) were added. The reaction was continued at 80°C for 5 hours. The reaction solution was analyzed by Pre-HPLC (formic acid) to obtain compound 38 (14 mg, yellow solid) in a yield of 12.1%. LCMS (ESI): m / z 453.40 [M+H] + ; RT = 3.245 min (6.00 min). 1 H NMR (400MHz, DMSO-d6): δ12.55(s,1H),11.14(s,1H),8.83(s,1H),8.52(d,J=4.8Hz,1H),7.86(d,J=8.0Hz,1H),7.56(d,J=8.4H z,1H),5.08-5.05(m,1H),3.80-3.19(m,8H),2.80(d,J=4.8Hz,3H),2.59-2.53(m,2H),1.72(d,J=6.8Hz,3H),1.21-1.73(m,3H).

[0195] Synthesis of compounds 38B1 and 38B2

[0196] Compound 38 (27 mg) was separated by chiral column, column type 250*25mm 10μm, mobile phase: methanol (+0.1% 7.0mol / L ammonia methanol), to obtain compound 38B1 (7.50mg) and compound 38B2 (3.12mg);

[0197] Compound 38B1: peak time 4.287 min; ee%>99, LCMS (ESI): m / z 453.5 [M+H] +;(DMSO-d6): δ12.23(s,1H),8.67(s,1H),8.39(d,J=4.8Hz,1H),7.87(s,1H),7.77(d,J=8.0Hz,1H),7.43(d,J=8.4Hz,1H),4.26-4.2 2(m,1H),2.89(s,4H),2.79(d,J=4.8Hz,3H),2.63(s,4H),2.58-2.54(m,2H),2.41(s,3H),1.46(d,J=6.4Hz,3H),1.20-1.16(m,3H).

[0198] Compound 38B2: peak time 5.411 min; ee%>98, LCMS (ESI): m / z 453.4 [M+H] + ; (DMSO-d6): δ12.22(s,1H),8.66(s,1H),8.40-8.37(m,1H),7.87(s,1H),7.76(d,J=8.0Hz,1H),7.43(d,J=8.0Hz,1H),4.28-4.22 (m,1H),2.89(s,4H),2.78(d,J=5.2Hz,3H),2.63(s,4H),2.56-2.54(m,2H),2.41(s,3H),1.46(d,J=6.8Hz,3H),1.20-1.16(m,3H).

[0199] Synthesis of compound 39

[0200] Step 1: Synthesis of 2-chloro-3-fluoro-5-iodopyridin-4-amine

[0201] Compound 39-1 (5 g, 3.12 mmol), N-iodosuccinimide (9.21 g, 40.94 mmol), and p-toluenesulfonic acid (0.29 g, 1.71 mmol) were dissolved in acetonitrile (50 mL) and stirred at 70°C for 16 hours. After cooling to room temperature, the reaction solution was poured into water (200 mL), filtered, and the solid was washed with saturated sodium sulfite solution and water, and dried under vacuum to obtain product 39-2 (9.1 g, 33.40 mmol, yield: 97.9%), as a white solid. LCMS (ESI): m / z 272.9 [M+H] + ; RT = 1.097 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ8.12(s,1H),6.68(s,2H).

[0202] Step 2: Synthesis of 7-chloro-3-ethyl-8-fluoro-3,4-dihydropyrido[3,4-b]pyrazin-2(1H)-one

[0203] Compound 39-2 (3.0 g, 11.01 mmol), 2-aminobutyric acid (2.27 g, 22.02 mmol), cuprous chloride (0.22 g, 2.20 mmol), and potassium phosphate (4.67 g, 22.02 mmol) were dissolved in dimethyl sulfoxide (30 mL). The reaction solution was stirred at 135°C under a nitrogen atmosphere for 16 hours. The mixture was cooled to room temperature, added with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to afford 39-3 (1.6 g, 6.97 mmol, yield: 63.28%, as a white solid). LCMS (ESI): m / z 230.1 [M+H] + ; RT = 0.910 min (2.50 min).

[0204] Step 3: Synthesis of 7-chloro-3-ethyl-8-fluoropyrido[3,4-b]pyrazin-2(1H)-one

[0205] Dissolve 39-3 (1.6 g, 6.97 mmol) and manganese dioxide (6.06 g, 69.67 mmol) in dichloromethane (25 mL). The reaction mixture was allowed to react at room temperature for 5 hours. Filter, collect the filtrate, and concentrate in vacuo to obtain the product 39-4 (1.4 g, 6.15 mmol, yield: 88.27%, white solid). LCMS (ESI): m / z 228.1 [M+H] + ; RT = 0.967 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ13.10 (s, 1H), 8.59 (s, 1H), 2.86-2.80 (m, 2H), 1.22 (t, J = 7.6Hz, 3H).

[0206] Step 4: Synthesis of 3-ethyl-8-fluoro-7-vinylpyrido[3,4-b]pyrazin-2(1H)-one

[0207] Compound 39-4 (650 mg, 2.86 mmol), tributyltin ethylene (1.36 g, 4.28 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (0.22 g, 0.29 mmol) were dissolved in dioxane (20 mL) and stirred at 80°C under nitrogen for 16 hours. After cooling to room temperature, the reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to afford 39-5 (400 mg, 1.82 mmol, yield: 63.90%), as a white solid. LCMS (ESI): m / z 220.1 [M+H] + ; RT = 1.067 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ12.89(s,1H),8.71(s,1H),7.07-7.00(m,1H),6.43(d ,J=9.6Hz,3H),6.67(d,J=6.4Hz,3H),2.86-2.80(m,2H),1.23(t,J=7.6Hz,3H).

[0208] Step 5: Synthesis of 7-(1,2-dihydroxyethyl)-3-ethyl-8-fluoropyrido[3,4-b]pyrazin-2(1H)-one

[0209] 39-5 (240 mg, 1.09 mmol), methanesulfonamide (0.16 g, 1.64 mmol), and AD-mixture-BETA (4.26 g, 5.47 mmol) were dissolved in tert-butanol (5 mL) and water (5 mL). The reaction mixture was allowed to react at room temperature for 16 hours. The filtrate was collected by filtration and concentrated to give the crude product 39-6 (0.25 g, 0.99 mmol, yield: 90.18%), as a white solid. LCMS (ESI): m / z 252.1 [MH] - ; RT = 0.849 min (2.50 min).

[0210] Step 6: Synthesis of compound 3-ethyl-8-fluoro-2-oxo-1,2-dihydropyrido[3,4-b]pyrazine-7-aldehyde

[0211] 39-6 (250 mg, 0.99 mmol) and sodium periodate (0.63 g, 2.96 mmol) were dissolved in tetrahydrofuran (5 mL) and water (5 mL) and reacted at room temperature for 16 hours. The mixture was filtered, the filtrate was collected, and concentrated to give the crude product. The crude product was purified by TLC (DCM:MeOH = 20:1) to give 39-7 (180 mg, 0.81 mmol, yield: 82.43%), as a white solid. LCMS (ESI): m / z 222.1 [M+H] + ; RT = 1.090 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ10.12(s,1H),8.55(s,1H),2.77-2.72(m,2H),1.19(t,J=7.2Hz,3H).

[0212] Step 7: 5-(4-((3-ethyl-8-fluoro-2-oxo-1,2-dihydropyrido[3,4-b]pyrazin-7-yl)methyl)piperazin-1-yl)-6-fluoro-N-methylpicolinamide

[0213] 39-7 (40 mg, 0.18 mmol) and 12a (51.71 mg, 0.22 mmol) were dissolved in dichloromethane (5 mL) and methanol (5 mL) and one drop of acetic acid was added. The reaction mixture was allowed to react at room temperature for 30 minutes, and then sodium cyanoborohydride (22.73 mg, 0.36 mmol) was added and the reaction continued for 30 minutes. Triethylamine (0.5 mL) was added to the reaction solution, which was concentrated and the residue was purified by HPLC (formic acid) to afford 39 (9.3 mg, 0.021 mmol, yield: 11.60%) as a white solid. LCMS (ESI): m / z 444.2 [M+H] + ; RT = 1.810 min (6 min). 1 H NMR (400MHz, DMSO-d6): δ8.69(s,1H),8.41-8.38(m,1H),7.83(d,J=8.0Hz,1H),7.53(t,J=8.0Hz,1H), 3.78(s,2H),3.14(s,4H),2.85-2.80(m,2H),2.76(d,J=4.8Hz,3H),2.64(s,4H),1.22(t,J=7.6Hz,3H).

[0214] Synthesis of compound 40

[0215] The synthesis method was the same as compound 39, except that L-alanine was used as the starting material instead of 2-aminobutyric acid. LCMS (ESI): m / z 430.2 [M+H]+ ; RT = 1.658 min (6 min). 1 H NMR (400MHz, DMSO-d6): δ8.66(s,1H),8.40(s,1H),7.83(d,J=8.0Hz,1H),7.56-7.5 2(m,1H),3.78(s,4H),3.14(s,4H),2.76(d,J=4.8Hz,3H),2.65(s,4H),2.42(s,3H).

[0216] Synthesis of compounds 41 and 42

[0217] According to the method described in compound 39, intermediate 12 was replaced with the intermediates in the following table to synthesize compounds 41 and 42. The structural formulas of the compounds in each example are shown in the previous table.

[0218] Synthesis of compound 43

[0219] Step 1: Synthesis of 4-amino-6-chloro-5-fluoronicotinonitrile

[0220] Compound 43-1 (5.0 g, 18.35 mmol), zinc cyanide (7.8 g, 23.86 mmol), and tetrakis(triphenylphosphine)palladium (1.1 g, 0.92 mmol) were dissolved in DMF (100 mL) and stirred at 100°C under a nitrogen atmosphere for 16 hours. The mixture was cooled to room temperature and extracted with ethyl acetate and water. The organic phase was collected, dried, and concentrated in vacuo. The residue was washed with ethyl acetate / petroleum ether (20 / 1) to afford the product 43-2 (3 g, yield: 95.28%), as a white solid. LCMS (ESI): m / z 170.1 [MH]-; RT = 0.841 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ8.20(s,1H),7.66(s,2H).

[0221] Step 2: Synthesis of methyl 4-amino-6-chloro-5-fluoronicotinate

[0222] 43-2 (1.1 g, 6.41 mmol) was dissolved in a solution of sulfuric acid (10 mL) and water (10 mL). The reaction solution was stirred at 120°C for 16 hours. After cooling to room temperature, the reaction solution was added to a saturated aqueous sodium bicarbonate solution and the solid was collected by filtration. The solid was purified by column chromatography (DCM) to give product 43-3 (0.8 g, 3.91 mmol, yield: 60.99%), white solid. LCMS (ESI): m / z 205.1 [M+H] +; RT = 0.977 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ8.38(s,1H),7.59(s,2H),3.86(s,3H).

[0223] Step 3: Synthesis of methyl 4-butyramido-6-chloro-5-fluoronicotinate

[0224] 43-3 (4.4 g, 21.51 mmol), 4-dimethylaminopyridine (2.63 g, 21.51 mmol), and DIEA (8.34 g, 64.52 mmol) were dissolved in DMF (50 mL). n-Butyryl chloride (4.58 g, 43.01 mmol) was added dropwise to the reaction solution at 0°C and allowed to react at room temperature for 16 hours. Extraction was performed with EA, and the organic phase was collected, dried, and concentrated. The residue was purified by column chromatography (DCM) to obtain the product 43-4 (1.3 g, yield: 22.01%, white solid). LCMS (ESI): m / z 275.1 [M+H] + ; RT = 1.087 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ10.51(s,1H),8.49(s,1H),3.78(s,3H)2.39(t,J=7.2Hz,2H),1.65-1.56(m,2H),0.93(t,J=7.6Hz,3H).

[0225] Step 4: Synthesis of 6-chloro-5-fluoro-4-(N-(4-methoxybenzyl)butyramido)nicotinate (

[0226] Compound 43-4 (0.40 g, 1.46 mmol), 4-methoxybenzyl chloride (0.34 g, 2.18 mmol), cesium carbonate (0.95 g, 2.91 mmol), and tetrabutylammonium bromide (0.23 g, 0.73 mmol) were dissolved in DMF (5 mL) and stirred at room temperature for 16 hours. The reaction solution was extracted with ethyl acetate / water, and the organic phase was collected, dried, and concentrated in vacuo. The residue was purified by column chromatography (EA:PE = 1:5) to afford compound 43-5 (400 mg, 1.01 mmol, yield: 69.07%) as a colorless, transparent oil. LCMS (ESI): m / z 395.2 [M+H] + ; RT = 1.437 min (2.50 min). 1H NMR (400MHz, DMSO-d6): δ8.69(s,1H),7.04(d,J=8.4Hz,2H),6.79(d,J=8.4Hz,2H),3.73(s, 2H),3.70(s,3H),3.66(s,3H),2.04-1.94(m,2H),1.52-1.47(m,2H),0.79(t,J=7.2Hz,3H).

[0227] Step 5: 7-Chloro-3-ethyl-8-fluoro-4-hydroxy-1-(4-methoxybenzyl)-1,6-naphthyridin-2(1H)-one

[0228] 43-5 (0.28 g, 0.71 mmol) was dissolved in tetrahydrofuran (20 mL) and sodium bis(trimethylsilyl)amide (0.39 g, 2.3 mmol) was added to the reaction solution at -78°C under a nitrogen atmosphere and stirred for 30 minutes. The reaction solution was quenched with water, extracted with ethyl acetate, and the aqueous phase was collected. The aqueous phase was adjusted to pH = 3, extracted with ethyl acetate, and the organic phase was dried and concentrated in vacuo to obtain 43-6 (140 mg, yield: 54.24%, white solid). LCMS (ESI): m / z 363.1 [M+H] + ; RT = 1.457 min (2.50 min). 1 H NMR (400MHz, DMSO-d6): δ8.76 (s, 1H), 7.03 (d, J = 8.4Hz, 2H), 6.86 (d, J = 8.4 Hz,2H),5.48(s,2H),3.71(s,3H),2.68-2.62(m,2H),1.06(t,J=7.2Hz,3H).

[0229] Step 6: 7-chloro-3-ethyl-8-fluoro-4-methoxy-1-(4-methoxybenzyl)-1,6-naphthyridin-2(1H)-one

[0230] Compound 43-6 (0.45 g, 1.24 mmol) and potassium carbonate (0.69 g, 4.96 mmol) were dissolved in DMF (20 mL). Iodomethane (0.44 g, 3.10 mmol) was added to the reaction solution at 0°C and stirred at room temperature for 16 hours. The reaction solution was extracted with ethyl acetate / water, and the organic phase was collected, dried, and concentrated in vacuo. The residue was purified by column chromatography (ethyl acetate:petroleum ether = 1:5) to afford compound 43-7 (200 mg, yield: 42.79%) as a white solid. 1H NMR (400MHz, DMSO-d6): δ8.66 (s, 1H), 7.07(d,J=8.4Hz,2H),6.88(d,J=11.6Hz,2H),5.49(s,2H),4.00(s,3H),3.71(s,3H)2.66-2.61(m,2H),1.15(t,J=7.2Hz,3H).

[0231] Step 7: Synthesis of 7-chloro-3-ethyl-8-fluoro-4-methoxy-1,6-naphthyridin-2(1H)-one

[0232] 43-7 (0.1 g, 0.29 mmol) was dissolved in methanesulfonic acid (2 mL) and stirred at room temperature for 1 hour. The reaction solution was added to water and filtered to collect a white solid to give 43-8 (60 mg, 0.23 mmol, yield: 88.09%), a white solid. LCMS (ESI): m / z 257.1 [M+H] + ; RT = 1.017 min (2.50 min). 1 H NMR (400MHz, CDCl3): δ9.58 (s, 1H), 8.50 (s, 1H), 3.95 (s, 3H), 2.66-2.61 (m, 2H), 1.61 (t, J = 7.6Hz, 3H).

[0233] Step 8: Synthesis of 3-ethyl-8-fluoro-4-methoxy-7-vinyl-1,6-naphthyridin-2(1H)-one

[0234] 43-8 (0.07 g, 0.27 mmol), tributyltin ethylene (0.13 g, 4.28 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (0.02 g, 0.27 mmol) were dissolved in dioxane (5 mL). The reaction solution was stirred at 80°C under a nitrogen atmosphere for 16 hours. After cooling to room temperature, the reaction solution was washed with ethyl acetate / petroleum ether (1 / 40) to obtain the crude product 43-9 (55 mg, yield: 81.25%), as a white solid. LCMS (ESI): m / z 249.1 [M+H] + ; RT = 1.127 min (2.50 min).

[0235] Step 9: Synthesis of 3-ethyl-8-fluoro-4-methoxy-2-oxo-1,2-dihydro-1,6-naphthyridine-7-aldehyde

[0236] 43-9 (55 mg, 0.22 mmol), methanesulfonamide (31.61 mg, 0.33 mmol), and AD-mixture-BETA (0.86 g, 1.11 mmol) were dissolved in tert-butanol (2.5 mL) and water (2.5 mL). The reaction solution was allowed to react at room temperature for 16 hours. The filtrate was collected and concentrated to give a yellow residue. The residue and sodium periodate (75.77 mg, 0.35 mmol) were dissolved in tetrahydrofuran (2.5 mL) and water (2.5 mL) and allowed to react at room temperature for 16 hours. The filtrate was collected and concentrated in vacuo to give the crude product. The crude product was purified by TLC (DCM:MeOH = 20:1) to give 43-10 (20 mg, 0.08 mmol, yield: 45.12%), as a white solid. LCMS (ESI): m / z 289.0 [MH] - ; RT = 1.129 min (2.50 min).

[0237] Step 10: Synthesis of 5-(4-((3-ethyl-8-fluoro-4-methoxy-2-oxo-1,2-dihydro-1,6-naphthyridin-7-yl)methyl)piperazin-1-yl)-6-fluoro-N-methylpicolinamide

[0238] Compound 43-10 (20 mg, 0.08 mmol) and compound 12a (28.57 mg, 0.12 mmol) were dissolved in methanol (5 mL) and one drop of acetic acid was added. After reacting at room temperature for 30 minutes, sodium cyanoborohydride (10.06 mg, 0.16 mmol) was added and the reaction continued for 30 minutes. The reaction solution was concentrated, and the residue was purified by HPLC (0.1% formic acid) to give compound 43 (1.24 mg, yield: 3.28%), as a white solid. LCMS (ESI): m / z 473.3 [M+H] + ; RT = 1.927 min (6 min). 1 H NMR (400MHz, DMSO-d6): δ12.21(s,1H),8.68(s,1H),8.40(d,J=4.8Hz,1H),7.83(d,J=8.0Hz,1H),7.55(t,J=10.4Hz,1H ),3.97(s,3H),3.78(s,2H),3.14(s,4H),2.76(d,J=4.4Hz,3H),2.65(s,4H),2.59-2.51(m,2H),1.11(t,J=7.6Hz,3H).

[0239] The stereoisomers of the above compounds can be obtained by conventional chiral resolution methods.

[0240] The present invention will be further described in detail below with reference to specific examples and data. It should be understood that these examples are merely illustrative of the present invention and are intended to illustrate the specific combinations, preparation methods, and functions and effects of the present invention, and are not intended to limit the scope of the invention in any way. The beneficial effects of the pharmaceutical combination of the present invention can also be determined by other test models known to those skilled in the relevant art.

[0241] Biological Examples

[0242] Experimental Example 1: Evaluation of the PARP1 / 2 inhibitory activity of compounds

[0243] The PARP1 / 2 inhibitory activity of the disclosed compounds was tested in an assay using histones as substrates.

[0244] Experimental purpose: According to established experimental methods, the IC50 values ​​of the compounds of the present application for inhibition of PARP1 / 2 enzyme activity were determined, with AZD-2281 (Olaparib) as a positive control compound.

[0245] Experimental reagents:

[0246] Recombinant human PARP1 protein (Abcam, cat. ab279663); recombinant human PARP2 protein (BPS, cat. 80502); recombinant histone H1 (Active Motif, cat. 81126); NAD+, Biotin-Labeled (BPS, cat. 80610); SuperBlock (TBS) Blocking Buffer (Thermo Scientific TM ,cat.37535);Streptavidin(HRP)(Abcam,cat.ab7403); Peroxidase Chemiluminescent Substrate Kit (Seracare, cat.5430-0040); 20xPBS (CST, cat.9808S); 20xPBST (CST, cat.9809S); AZD2281 (Selleck, cat.S1060)

[0247] Experimental methods:

[0248] 1. Compound preparation: Dilute the compound with DMSO to a 1000-fold final concentration in a 384-well plate and set aside.

[0249] 2. Coating microplate:

[0250] 1) Dilute Histone with PBS, add 25 μL of Histone mixture to each well, and incubate for 2 hours. 2) Wash each well five times with PBST. Remove the solution on a clean paper towel. 3) Add 75 μL of Blocking Buffer to each well and incubate at room temperature for 1 hour. 4) Wash each well five times with PBST. Remove the solution on a clean paper towel.

[0251] 3. Ribosylation reaction:

[0252] 1) Transfer 25 nL of the reserved compound at a 1000x final concentration to a 384-well plate. Add 25 nL of 100% DMSO to each of the Min and Max control wells. 2) Prepare a PARP1 or PARP2 solution at a 2.5x final concentration in 1x Assay buffer. 3) Add 10 μL of enzyme solution to each of the compound wells and Max control wells; add 10 μL of 1x Assay buffer to the Min control wells. 4) Centrifuge at 1000 rpm for 60 seconds and incubate at room temperature for 15 minutes. 5) Prepare a substrate solution at a 1.67x final concentration in 1x Assay buffer and add 15 μL to each well to initiate the reaction. 6) Centrifuge at 1000 rpm for 60 seconds and incubate at room temperature for 2 hours. 7) Wash each well five times with PBST. Drain the solution on a clean paper towel.

[0253] 4. Detection:

[0254] 1) Prepare Streptavidin-HRP solution, add 25 μL to each well, centrifuge at 1000 rpm for 60 seconds, and incubate at room temperature for 30 minutes. 2) Wash each well five times with PBST. Drain the solution on a clean paper towel. 3) Add 50 μL of ELISA Chemiluminescent Substrate to each well. 4) Centrifuge at 1000 rpm for 60 seconds. Read the plate using EnSight after 5 minutes.

[0255] 5. Data Analysis:

[0256] The inhibition rate was calculated using the following formula: inhibition rate % = (maximum signal - compound signal) / (maximum signal - minimum signal) × 100, where "minimum signal" is the mean value of the negative control wells and "maximum signal" is the mean value of the positive control wells.

[0257] Fitting a dose-effect curve: Using the log concentration as the X-axis and the percent inhibition as the Y-axis, the log(inhibitor) vs. response - variable slope function of the GraphPad Prism 5 analysis software was used to fit the dose-effect curve to obtain the IC50 value of the disclosed compounds for enzyme activity inhibition. The fitting formula is: Y = Base + (Top - Base) / (1 + 10^((logIC50 - X) * HillSlope))

[0258] Table 1. IC values ​​of the compounds of the present disclosure for inhibition of PARP1 / 2 enzymes 50

[0259] *In the fifth step of the glycosylation reaction in the PARP1 enzyme activity assay, 500 μM NAD was added to the substrate solution. + .

[0260] It can also be seen from the results in Table 1 that the compounds disclosed herein have a very significant selective inhibitory effect on PARP1 compared with the compounds in the prior art.

[0261] Example 2: MDA-MB-436 cell proliferation inhibition test

[0262] Human breast cancer MDA-MB-436 (purchased from ATCC) cells were cultured in DMEM medium (supplemented with 10% fetal bovine serum and 1% double antibody) at 37°C and 5% carbon dioxide. Cells in the logarithmic growth phase were taken, digested, and a cell suspension of a certain concentration was prepared. The cell suspension was inoculated into a 96-well plate, and 100 μL of cell suspension was added to each well of the 96-well plate. After incubation overnight, different concentrations of compounds were added and placed in a cell culture incubator for 7 days. After the culture was completed, 50 μL of CellTiter-Glo reagent was added to each well, mixed with a microplate shaker for 2 minutes, and placed at room temperature for 60 minutes. The fluorescence value is read by the multimode microplate reader according to the formula: [(1-(RLU compound -RLU blank ) / (RLU control -RLU blank )) × 100%] to calculate the cell proliferation inhibition rate. GraphPad Prism 6.0 software was used to fit the IC 50 value.

[0263] Table 2. MDA-MB-436 cell proliferation inhibitory activity of the compounds disclosed herein

[0264] The experimental results show that the compound of the present invention has significant proliferation inhibitory activity on MDA-MB-436 cells.

[0265] Experimental Example 3: Preliminary pharmacokinetic testing experiment

[0266] 1. Healthy ICR mice were randomly divided into 3 groups of 3 mice each, with 9 male mice weighing 30-35 g for each administration route, and the test compound was administered intravenously (1 mg / kg) or orally (5 mg / kg).

[0267] The animals were fasted for 12 hours before the experiment and allowed to drink water freely. They were fed 4 hours after the administration.

[0268] 2. Blood collection time and sample processing

[0269] Intravenous and oral administration: 0.25h, 0.5h, 1.0h, 2.0h, 3.0h, 4.0h, 6.0h, 8.0h and 24h after administration.

[0270] Blood was collected continuously, with 3 animals collected at each time point. Plasma collection and processing: 30-40 μL of venous blood was collected from the retroorbital venous plexus of mice at the above-set time points, placed in EDTA-K2 tubes, centrifuged at 3500 rpm for 10 minutes, and plasma was separated and frozen in a -20°C refrigerator.

[0271] 3. Sample testing and data analysis

[0272] The concentration of the compound in mouse plasma was determined by LC / MS / MS, and the pharmacokinetic parameters after administration were calculated using a non-compartmental model using Phoenix 8.3 software (Pharsight, USA).

[0273] 4. Experimental Results

[0274] Table 3. Pharmacokinetic parameters of the compounds of the present disclosure in mouse plasma

[0275] The experimental results show that the pharmacokinetics of the compound of the present invention in mice after oral administration show a long half-life T 1 / 2 , higher in vivo exposure AUC 0-t .

[0276] Example 4: Evaluation of bidirectional permeability using the MDR1-MDCKⅡ cell model

[0277] MDR1-MDCKⅡ cells were expressed at 3.3×10 5Cells were seeded into 96-well plates at a concentration of 10 cells / mL and grown for 4-7 days to form a monolayer of confluent cells. The test compound was added at a concentration of 2 μM to the dosing end wells on the apical or basolateral side of the monolayer cells and incubated at 37.0°C in a 5.0% CO2 incubator for 2.5 hours. The integrity of the cell monolayer was determined by the fluorescein exclusion assay. The buffer was removed from the apical and basolateral sides, and the concentration of the test compound was determined using LC-MS / MS. The concentration data was used to calculate the apparent permeability coefficient of transport from the apical side to the basolateral side and from the basolateral side to the apical side of the monolayer cells, and to calculate the efflux rate. Calculation formula: Efflux Ratio = Papp (BA) / Papp (AB)

[0278] Table 4. Bidirectional permeability of compounds of the present disclosure to MDR1-MDCKII cells

[0279] The experimental results show that the compound of the present invention has higher cell permeability and lower efflux rate in MDR1-MDCKⅡ cells.

Claims

1. A compound represented by general formula (I), or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof, in, X 1 、X 2 Each independently selected from N and C(H), C(OCH3); X 3 Selected from N and CR 10 , where R 10 is hydrogen or a halogen; R 1 Selected from hydrogen, C1-C6 alkoxy, unsubstituted or halogen-substituted C1-C6 alkyl, unsubstituted or halogen-substituted C3-C6 cycloalkyl; R 2 is hydrogen, halogen or C1-C6 alkyl; R 3 、R 4 、R 4‘ 、R 5 、R 6 、R 7 Each is independently hydrogen or C1-C6 alkyl; R 8 is selected from hydrogen, halogen, unsubstituted or halogen-substituted C1-C6 alkyl; R 9 is selected from hydrogen, C3-C6 cycloalkyl, unsubstituted or halogen-substituted C1-C6 alkyl; m is 1 or 2; The conditions are: When X 1 is N, and X 2 and X 3 When it is C(H), then R 3 、R 4 、R 4‘ 、R 5 、R 6 、R 7 are not simultaneously hydrogen; and When R 2 When it is H, then R 3 、R 4 、R 4‘ 、R 5 、R 6 、R 7 Not hydrogen at the same time.

2. The compound according to claim 1, or its stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug, wherein: R 10 is hydrogen or fluorine; R 1 is selected from hydrogen, C1-C4 alkoxy, unsubstituted or halogen-substituted C1-C4 alkyl; R 2 is hydrogen, halogen or C1-C4 alkyl; R 3 、R 4 、R 4‘ 、R 5 、R 6 、R 7 Each is independently hydrogen or C1-C4 alkyl; R 8 is selected from hydrogen, halogen, unsubstituted or halogen-substituted C1-C4 alkyl; R 9 is hydrogen, C3-C6 cycloalkyl or C1-C4 alkyl; The other substituents are the same as defined in claim 1.

3. The compound according to claim 1 or 2, or its stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug, wherein: m is 1; R 1 is hydrogen, C1-C2 alkoxy, C1-C4 alkyl or C1-C4 fluoroalkyl; R 2 is hydrogen or fluorine; R 8 is selected from hydrogen, halogen, C1-C4 alkyl and C1-C4 fluoroalkyl; R 9 is hydrogen or C1-C4 alkyl; The other substituents are the same as defined in claim 1.

4. The compound according to claim 1, or its stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug, wherein: X 1 is selected from N, C(H) and C(OCH3); X 2 is C(H); X 3 is N; R 2 is hydrogen or F; The other substituents are the same as defined in claim 1.

5. The compound of claim 1, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate or prodrug thereof, in, In the structure of formula (I) Partially selected from the following structures: The other substituents are the same as defined in claim 1.

6. The compound according to claim 1, or its stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug, wherein: The compound represented by formula (I) has a structure represented by formula (I-1) to formula (I-8) or a racemic form thereof: Each substituent has the same meaning as defined in claim 1.

7. The compound according to claim 1, or its stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug, wherein: The compound of formula (I) is selected from the following specific compounds:

8. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 7, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate or prodrug thereof, and a pharmaceutically acceptable carrier.

9. Use of the compound of formula (I) according to any one of claims 1 to 7, or its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, crystal forms, solvates, hydrates or prodrugs, or the pharmaceutical composition according to claim 8 in the preparation of a medicament for preventing, treating or ameliorating a disease by inhibiting PARP1. in, The disease is cancer, The cancer genome is of a homologous recombination repair deficient type, Alternatively, the cancer is dependent on a pathway that repairs double-strand DNA damage but lacks homologous recombination, Alternatively, the cancer comprises one or more cancer cells that lack the ability to repair DNA double-strand breaks by homologous recombination relative to normal cells, Alternatively, the cancer comprises one or more cancer cells that lack BRCA1 or BRCA2, or that have a BRCA1 or BRCA2 mutation.

10. The use according to claim 9, wherein The cancer includes any one of ovarian cancer, breast cancer, fallopian tube cancer, endometrial cancer, peritoneal cancer, gastric cancer, colon cancer, bladder cancer, pancreatic cancer, biliary tract cancer, osteosarcoma, cervical cancer, head and neck tumors, germ cell and embryonal cancers, esophageal cancer, glioblastoma, Ewing sarcoma, pancreatic cancer, melanoma, bile duct cancer, prostate cancer, small cell lung cancer, non-small cell lung cancer, lymphoma and blood cancer.

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