Phthalazinone compound as well as preparation method, pharmaceutical composition and application thereof

Abstract

The invention discloses a phthalazinone compound as shown in a formula I as well as a preparation method, a pharmaceutical composition and application of the phthalazinone compound. The compound has excellent inhibitory activity on poly (adenosine diphosphate ribose polymerase) and nicotinamide phosphoribose transferase, has remarkable inhibitory activity on BRAC1 / 2 mutation and BRAC1 / 2 non-mutation tumors, and is relatively small in toxic and side effects and good in safety. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a phthalazinone compound and a preparation method, a pharmaceutical composition and application thereof, and in particular to a phthalazinone compound having dual inhibitory activities of poly(ADP-ribose) polymerase inhibitor and nicotinamide phosphoribosyltransferase and a preparation method, a pharmaceutical composition and application thereof. Background Art

[0002] Poly (ADP-ribose) polymerase (PARP) can catalyze the transfer of ADP-ribose of nicotinamide adenine dinucleotide to receptor proteins, thereby catalyzing the poly (ADP-ribosylation) of proteins. Therefore, PARP plays a key role in single-strand DNA repair. The PARP protein family consists of 18 members, and their catalytic domains show a high degree of similarity. Broken single-stranded DNA can be repaired with the participation of PARP, and the cell can survive. When PARP is inhibited by its inhibitor, the unrepaired single-stranded DNA will form double-stranded DNA breaks. Under normal circumstances, the homologous recombination repair mechanism can repair the broken double-stranded DNA, and the cell can continue to survive. However, when there is a homologous recombination repair defect in the cell, that is, the lack of related enzymes involved in the HR repair pathway, such as BRAC1 / 2, the cell dies because the DNA damage cannot be repaired. This is the synthetic lethal mechanism of PARP inhibitors and HRD.

[0003] NAD + It is a cofactor of many oxidoreductases in the body, thus participating in the energy metabolism process in the body, and is also a substrate for key enzymes in many signal transduction processes. The salvage synthesis pathway using nicotinamide as the starting material is a way to replenish NAD in the body. + The most important and direct source of NAD, and nicotinamide phosphoribosyltransferase is the key rate-limiting enzyme that catalyzes this process. Tumor cell proliferation and energy requirements are much greater than normal cells, so tumor cells have a high demand for NAD + In addition to participating in energy metabolism, NAMPT plays a key role in cancer stem cells, epithelial-mesenchymal transition and metastasis, DNA repair, signal transduction, and tumor immune escape.

[0004] Currently, olaparib, rucaparib, niraparib, talazoparib, fluzoparib and pamiparib, as PARP small molecule inhibitors, have been used to treat ovarian cancer, breast cancer, pancreatic cancer and prostate cancer carrying BRAC mutations. Although PARP inhibitors have significantly improved the prognosis of a variety of HRD-positive malignant tumors, the indication range of PARP inhibitors is narrow, the patient population that benefits is limited, and it is only effective for HRD-positive tumors that can induce synthetic lethality, which limits the clinical application of PARP inhibitors. At the same time, a large proportion of HRD-positive tumors also show innate resistance to PARP inhibitors, or develop acquired resistance after a period of medication. In addition, FK866 and CHS828, as small molecule inhibitors of NAMPT, were terminated in clinical studies due to poor therapeutic effects and dose-dependent toxicity. Summary of the invention

[0005] Purpose of the invention: The first purpose of the present invention is to provide a phthalazinone compound capable of inhibiting the activity of poly(ADP-ribose) polymerase and nicotinamide phosphoribosyltransferase; the second purpose is to provide a method for preparing the compound; the third purpose is to provide a pharmaceutical composition containing the compound; and the fourth purpose is to provide a pharmaceutical application of the compound and its pharmaceutical composition.

[0006] Technical solution: The phthalazinone compound of the present invention as shown in formula I or its pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, prodrug, solvate, deuterated substance, hydrate,

[0007]

[0008] In Formula I:

[0009] X is N or C;

[0010] L1 is a single bond or

[0011] Y is C 6-10 Aryl or 5-10 membered heteroaryl containing 1-3 N, O, S heteroatoms;

[0012] L2 is -(CH2) n -, n is 0, 1, 2 or 3;

[0013] R1 is a 9- to 10-membered bicyclic heteroaryl group containing 1 N and 1 to 2 N, O, and S heteroatoms.

[0014] The compounds of the present invention may exist in specific stereoisomeric forms. The term "isomer" refers to isomers with identical structures but different arrangements of atoms in space. It includes cis and trans (or Z and E) isomers, (-)- and (+)-isomers, (R)- and (S)-enantiomers, diastereomers, (D)- and (L)-isomers, tautomers, atropisomers, conformers and mixtures thereof (such as racemates, mixtures of diastereomers). Substituents in the disclosed compounds may have additional asymmetric atoms. All of these stereoisomers and their mixtures are included within the scope of the present disclosure. Optically active (-)- and (+)-isomers, (R)- and (S)-enantiomers and (D)- and (L)-isomers may be prepared by chiral synthesis, chiral reagents or other conventional techniques. An isomer of a compound disclosed herein can be prepared by asymmetric synthesis or chiral auxiliary, or, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereoisomer salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art to obtain pure isomers. In addition, the separation of enantiomers and diastereoisomers is usually completed by chromatography.

[0015] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared from compounds having specific substituents discovered by the present invention with relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the free form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the free form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid (forming carbonates or bicarbonates), phosphoric acid (forming phosphates, monohydrogen phosphates, dihydrogen phosphates, sulfuric acid (forming sulfates or bisulfates), hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, Citric acid, tartaric acid and methanesulfonic acid and similar acids; organic acid salts also include amino acids (such as arginine, etc.), glucuronic acid and other organic acid salts. Certain specific compounds of the present invention contain basic and acidic functional groups, and can be converted into any base or acid addition salt. Preferably, the salt is contacted with a base or acid in a conventional manner, and the parent compound is separated, thereby regenerating the free form of the compound. The free form of the compound differs from its various salt forms in certain physical properties, such as different solubility in polar solvents.

[0016] The "pharmaceutically acceptable salts" of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. In general, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of an appropriate base or acid 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.

[0017] The term "metabolite" refers to a pharmaceutically active product produced by the in vivo metabolism of a compound of Formula I or a salt thereof. Such a product may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, glucuronidation, enzymatic cleavage, etc. of the administered compound. Therefore, the present invention includes metabolites of the compounds of the present invention, including compounds produced by a process of contacting a compound of the present invention with a mammal for a period of time sufficient to obtain a metabolite thereof.

[0018] Metabolites are typically identified by preparing radiolabeled isotopes of the compounds of the invention, administering them parenterally to animals, such as rats, mice, guinea pigs, monkeys, or humans, at detectable doses (e.g., greater than about 0.5 mg / kg), allowing sufficient time for metabolism to occur (typically about 30 seconds to 30 hours), and isolating the conversion products from urine, blood, or other biological samples. These products are easily isolated because they are labeled (others are isolated by using antibodies that can bind to antigenic epitopes present in the metabolites). The metabolite structure is determined in a conventional manner, for example, by MS, LC / MS or NMR analysis. Generally, the analysis of metabolites is performed in the same manner as conventional drug metabolism studies known to those skilled in the art. As long as the metabolite products are not otherwise not found in vivo, they can be used in assays for therapeutic dosing of the compounds of the invention. The compounds of the invention may contain non-natural proportions of atomic isotopes on one or more of the atoms that make up the compound. For example, compounds may be labeled with radioisotopes, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C) All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

[0019] In addition to the form of salts, the compounds provided by the present invention also exist in the form of prodrugs. The prodrugs of the compounds described herein are easily chemically changed under physiological conditions to be converted into the compounds of the present invention. Any compound that can be converted in vivo to provide a bioactive substance (i.e., a compound shown in Formula I) is a prodrug within the scope and spirit of the present invention. For example, a carboxyl-containing compound can form a physiologically hydrolyzable ester, which acts as a prodrug by being hydrolyzed in vivo to obtain the compound shown in Formula I itself. The prodrug is preferably administered orally, because hydrolysis occurs mainly under the influence of digestive enzymes in many cases. When the ester itself is active or hydrolysis occurs in the blood, parenteral administration can be used.

[0020] The term "deuterated compound" means that any hydrogen atom in a molecule can be replaced by its isotope deuterium. Any hydrogen atom in the compound of the present invention can be replaced by a deuterium atom.

[0021] The term "aryl" refers to an aromatic group having a specified number of carbon atoms, preferably a monocyclic, bicyclic or tricyclic aromatic group, and when it is a bicyclic or tricyclic aromatic group, each ring satisfies the Huckel rule. 6-10 The aryl group refers to an aromatic group containing 6 to 10 carbon atoms, such as phenyl.

[0022] The term "heteroaryl" refers to a 9-10 membered bicyclic heteroaryl containing one N and one or two other heteroatoms selected from N, O and S, including a bicyclic ring formed by condensing an unsaturated monocyclic ring containing heteroatoms with an aryl group, a bicyclic ring formed by condensing an unsaturated monocyclic ring containing heteroatoms with an unsaturated monocyclic ring containing heteroatoms, and a bicyclic ring formed by condensing a saturated monocyclic ring containing heteroatoms with a heteroaryl or aryl group. When the "heteroaryl" is a bicyclic ring formed by condensing a saturated monocyclic ring containing heteroatoms with a heteroaryl group, it is connected to other fragments and groups in the compound shown in Formula I through a saturated monocyclic ring containing a heterocyclic ring. The 9-10 membered bicyclic ring includes, but is not limited to, imidazopyridine, imidazopyrimidine, pyrazolopyridine, thienopyridine, furopyridine, tetrahydropyrrolopyridine or tetrahydropyrrolophenyl.

[0023] It will be understood by those skilled in the art that the structural formulas used in the present invention to describe groups are based on the conventions used in the art. It means that the corresponding group is connected to other fragments and groups in the compound shown in Formula I through this site.

[0024] Preferably, in Formula I:

[0025] Y is phenyl or pyridyl;

[0026] n is 0, 1 or 2.

[0027] Preferably, in Formula I:

[0028] Y is phenyl,

[0029] n is 0 or 1.

[0030] Preferably, in Formula I:

[0031] R1 is pyridoimidazolyl, pyridopyrazolyl, pyridothiphenyl, pyridofuranyl, pyridotetrahydropyrrolyl or phenyltetrahydropyrrolyl.

[0032] Preferably, in Formula I:

[0033] R1 is

[0034]

[0035] Preferably, the compound of the present invention as shown in formula I is selected from any one of the following compounds:

[0036]

[0037] NAMPT inhibitors inhibit NAD +The production of PARP inhibits the substrate supply of PARP and increases the competitive inhibition of PARP inhibitors on the PARP catalytic domain. Therefore, NAMPT inhibitors can increase DNA damage, thereby increasing the anti-tumor activity of PARP inhibitors. NAMPT activity can reduce the expression of BRAC1 / 2, reduce the expression of RAD51, and induce HRD. NAMPT inhibitors and PARP1 inhibitors exert synergistic anti-tumor effects in BRAC wild-type triple-negative breast cancer and Ewing sarcoma. However, combined drug use is prone to drug-drug interactions, unclear pharmacokinetic properties, and poor patient compliance. Dual-target or multi-target drugs can reduce toxicity, increase efficacy, and avoid the defects of combined drug use. Therefore, the development of PARP and NAMPT dual-target inhibitors can improve the narrow indications of PARP inhibitors, reduce the large toxic and side effects of NAMPT inhibitors, and drug resistance.

[0038] In view of the limited use of poly(ADP-ribose) polymerase inhibitors and the dose-dependence of nicotinamide phosphoribosyltransferase inhibitors, the present invention designs a phthalazinone poly(ADP-ribose) polymerase inhibitor and nicotinamide phosphoribosyltransferase dual inhibitor, which shows significant therapeutic effects on tumors carrying BRAC1 / 2 mutations and tumors without BRAC1 / 2 mutations, and has less toxic side effects.

[0039] The method for preparing the phthalazinone compound as shown in Formula I of the present invention or its pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformer, metabolite, prodrug, solvate, deuterated product, hydrate comprises the following steps:

[0040] The compound of formula II and the compound of formula III undergo condensation reaction under the action of a base and a condensing agent to obtain the compound of formula I:

[0041]

[0042] Wherein, X, Y, L1, L2, R1 are as described above;

[0043] The solvent described in the above reaction includes but is not limited to N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, dichloromethane, N-methylpyrrolidone or a mixed solvent thereof;

[0044] The base described in the above reaction is an organic base, including but not limited to triethylamine, pyridine, N,N-diisopropylethylamine (DIPEA); preferably triethylamine or N,N-diisopropylethylamine;

[0045] The condensing agent described in the above reaction includes but is not limited to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU), 1-hydroxybenzotriazole (HOBt), 1-hydroxy-7-azobenzotriazole (HOAt), O -benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP) or benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP); preferably EDCI, HATU, BOP or PyBOP.

[0046] The phthalazinone compounds of the present invention as shown in formula I or pharmaceutically acceptable salts, enantiomers, diastereomers, cis-trans isomers, tautomers, atropisomers, conformers, metabolites, prodrugs, solvates, deuterated substances, and hydrates thereof are used in the preparation of drugs for poly(ADP-ribose) polymerase inhibitors and / or drugs for nicotinamide phosphoribosyltransferase inhibitors.

[0047] The phthalazinone compounds of the present invention as shown in formula I or their pharmaceutically acceptable salts, enantiomers, diastereomers, cis-trans isomers, tautomers, atropisomers, conformers, metabolites, prodrugs, solvates, and hydrates are used in the preparation of drugs for treating and / or preventing cancer.

[0048] Preferably, the drug is a drug for treating and / or preventing one or more of bone cancer, lung cancer, gastric cancer, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, glioma, prostate cancer, pancreatic cancer, liver cancer, brain cancer, bladder cancer, cervical cancer, testicular cancer, kidney cancer, head and neck cancer, thyroid cancer, esophageal cancer, lymphoma, leukemia or skin cancer.

[0049] The pharmaceutical composition of the present invention contains a phthalazinone compound as shown in formula I or a pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, prodrug, solvate, deuterated substance, hydrate thereof and a pharmaceutically acceptable carrier or excipient.

[0050] Preferably, in the pharmaceutical composition, the amount of the phthalazinone compound or its pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, solvate, deuterated substance, hydrate thereof is a therapeutically effective amount.

[0051] The pharmaceutical composition of the present invention is used in the preparation of drugs for poly(ADP-ribose) polymerase inhibitors and / or drugs for nicotinamide phosphoribosyltransferase inhibitors.

[0052] The pharmaceutical composition of the present invention is used in preparing drugs for treating and / or preventing cancer.

[0053] Preferably, the cancer drug is a drug for treating and / or preventing one or more of bone cancer, lung cancer, gastric cancer, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, glioma, prostate cancer, pancreatic cancer, liver cancer, brain cancer, bladder cancer, cervical cancer, testicular cancer, kidney cancer, head and neck cancer, thyroid cancer, esophageal cancer, lymphoma, leukemia or skin cancer.

[0054] The pharmaceutically acceptable carrier can be an adjuvant widely used in the field of drug production. The adjuvant is mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide a method to dissolve the active ingredient at a desired rate after the subject receives the administration, or promote the effective absorption of the active ingredient after the subject receives the composition. The pharmaceutical adjuvant can be an inert filler, or provide a certain function, such as stabilizing the overall pH value of the composition or preventing the degradation of the active ingredient of the composition. The pharmaceutical adjuvant can include one or more of the following adjuvants: adhesives, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesive agents, glidants, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents and sweeteners.

[0055] The pharmaceutical composition of the present invention can be prepared according to the disclosed content using any method known to those skilled in the art, such as conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding or lyophilizing processes.

[0056] The pharmaceutical composition of the present invention can be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid preparations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intraarterial, intramuscular) administration. The pharmaceutical composition of the present invention can also be a controlled release or sustained release dosage form (e.g., liposomes or microspheres). Examples of solid oral preparations include, but are not limited to, powders, capsules, caplets, soft capsules, and tablets. Examples of liquid preparations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs, and solutions. Examples of topical preparations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum preparations. Examples of preparations for parenteral administration include, but are not limited to, solutions for injection, dry powder preparations that can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic preparations; aerosols, such as nasal sprays or inhalers; liquid dosage forms suitable for parenteral administration; suppositories and lozenges.

[0057] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0058] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0059] The compounds designed in the present invention have excellent inhibitory activity against poly(ADP-ribose) polymerase and nicotinamide phosphoribosyltransferase, and have significant inhibitory activity against tumors with BRAC1 / 2 mutations and tumors without BRAC1 / 2 mutations, and have less toxic and side effects and good safety. DETAILED DESCRIPTION

[0060] The technical solution of the present invention is further described below in conjunction with embodiments.

[0061] Example 1: Synthesis of Compound S1

[0062]

[0063] Step 1: Synthesis of intermediate 7

[0064]

[0065] a) Compound 1 (3.5 g, 11.25 mmol) was dissolved in acetonitrile (34 mL), 4-methoxythiophenol (2.58 g, 13.73 mmol) and K2CO3 (2.33 g, 16.88 mmol) were added to the reaction system, and the reaction was carried out at room temperature for 6 hours. After the reaction was complete, K2CO3 was removed by filtration, and the mixture was concentrated under reduced pressure and purified by column chromatography to obtain intermediate 2 (2.95 g, 71%). MS (ESI, m / z): 372 (M + +1).

[0066] b) Intermediate 2 (2.95 g, 7.95 mmol) was dissolved in DCM (25 mL), 3-chloroperoxybenzoic acid (6.86 g, 39.75 mmol) was added at 0°C, and the mixture was transferred to room temperature for reaction for 2 hours. After the reaction was complete, the reaction mixture was cooled to 0°C, and saturated sodium thiosulfate solution (80 mL) was slowly added to quench the mixture, and the mixture was extracted with dichloromethane (30 mL×3). The organic phases were combined, washed twice with 1M NaOH aqueous solution, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain Intermediate 3 (2.78 g, 87%). MS (ESI, m / z): 404 (M + +1).

[0067] c) Intermediate 3 (2.78 g, 6.90 mmol) was dissolved in DMF (28 mL), tetrakis(triphenylphosphine)palladium (797 mg, 0.69 mmol) and zinc cyanide (956 mg, 8.14 mmol) were added to the reaction system, and the mixture was reacted at 85° C. for 7 hours. After the reaction was complete, the mixture was diluted with water, filtered, and the filter cake was purified by column chromatography to obtain intermediate 4 (1.5 g, 62%). MS (ESI, m / z): 351 (M + +1).

[0068] d) Intermediate 4 (1.5 g, 4.29 mmol) was dissolved in MeOH / THF (13 mL / 13 mL), nickel chloride hexahydrate (1.02 g, 4.29 mmol) was added to the reaction system, sodium borohydride (243 mg, 6.44 mmol) was slowly added at 0°C, the temperature was raised to 60°C for reaction for 12 hours, and after the reaction was complete, the filtrate was filtered with diatomaceous earth, the filtrate was cooled to 0°C, 2M HCl aqueous solution was added to quench the sodium borohydride, and the sodium borohydride was extracted with ethyl acetate (20 mL×3), the organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain intermediate 5 (600 mg, 40%). MS (ESI, m / z): 355 (M + +1).

[0069] e) Intermediate 5 (120 mg, 0.34 mmol) was dissolved in DMF (1 mL), and imidazo[1,2-A]pyridine-7-carboxylic acid (55 mg, 0.34 mmol), BOP (225 mg, 0.51 mmol) and DIPEA (131 mg, 1.02 mmol) were added to the reaction system. The reaction was allowed to react at room temperature for 16 hours. After the reaction was complete, water was added to dilute the mixture, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain Intermediate 6 (140 mg, 83%). MS (ESI, m / z): 499 (M + +1).

[0070] f) Intermediate 6 (140 mg, 0.28 mmol) was dissolved in DCM (1 mL), TFA (0.5 mL) was added to the reaction system, and the reaction was allowed to react at room temperature for 1 hour. After the reaction was complete, dichloromethane was removed by distillation under reduced pressure, water was added to dilute, 2M NaOH aqueous solution was added to adjust the pH to 8-10 at 0°C, and dichloromethane was extracted. The organic phases were combined, dried over anhydrous Na2SO4, filtered to remove salt, and concentrated under reduced pressure to obtain Intermediate 7 (45 mg, 41%). MS (ESI, m / z): 399 (M + +1).

[0071] Step 2: Synthesis of compound S1

[0072]

[0073] a) Intermediate 7 (45 mg, 0.11 mmol) and compound 8 (21 mg, 0.11 mmol) were dissolved in DMF (1 mL), and BOP (75 mg, 0.17 mmol) and DIPEA (44 mg, 0.34 mmol) were added to the reaction system. The mixture was reacted at room temperature for 12 hours. After the reaction was complete, the mixture was diluted with water and extracted with ethyl acetate (10 mL×3). The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain S1 (41 mg, 53%).

[0074] S1: 1HNMR (400MHz, DMSO-d6) δ12.58(s,1H),9.27(t,J=5.9Hz,1H),9.18(s,1H),8.26(d,J=7.6Hz,1H),8.09(s,1H),7.97( d,J=7.9Hz,1H),7.88(t,J=7.3Hz,1H),7.83(dd,J=8.1,2.7Hz,3H),7.73-7.61(m,5H),7.45-7.37(m,1H),7.33(s,1H) ,7.20(t,J=9.0Hz,1H),4.63(d,J=5.8Hz,2H),4.55(d,J=13.2Hz,1H),4.32(s,2H),3.56(t,J=12.1Hz,1H),3.01(t,J= 13.0Hz,1H),2.75(t,J=12.6Hz,1H),1.93(d,J=12.9Hz,1H),1.77(s,1H),1.41(d,J=12.9Hz,2H).MS(ESI,m / z):679(M + +1).

[0075] Example 2: Synthesis of Compound S2

[0076]

[0077] The synthesis method is similar to that of Example 1, and compound S2 can be obtained by simply replacing the corresponding raw materials.

[0078] S2: 1HNMR (400MHz, DMSO-d6) δ13.93(s,1H),12.60(s,1H),9.37(t,J=6.0Hz,1H),9.04(d,J=2.1Hz,1H),8.78(d,J=2.1Hz,1H),8.31(s ,1H),8.30-8.21(m,1H),7.96(d,J=7.9Hz,1H),7.91-7.78(m,4H),7.65(d,J=8.0Hz,2H),7.44-7.37(m,1H),7.33(s,1H),7.20(t ,J=9.0Hz,1H),4.65(d,J=5.8Hz,2H),4.55(d,J=13.0Hz,1H),4.32(s,2H),3.56(t,J=23.1Hz,1H),3.01(t,J=26.7,12.5Hz,1H), 3.01(d,J=25.7Hz,1H),2.75(t,J=12.5Hz,1H),1.93(d,J=12.6Hz,1H),1.77(s,1H),1.39(d,J=23.7Hz,2H).MS(ESI,m / z):680(M + +1).

[0079] Example 3: Synthesis of Compound S3

[0080]

[0081] Step 1: Synthesis of intermediate 10

[0082]

[0083] a) Intermediate 5 (612 mg, 1.72 mmol), N,N'-disuccinimidyl carbonate (530 mg, 2.07 mmol) and pyridine (136 mg, 1.72 mmol) were dissolved in anhydrous acetonitrile (8.6 mL), and 2,3-dihydro-1H-pyrrolo[3,4-C]pyridine solution was added dropwise at 0°C. 2,3-dihydro-1H-pyrrolo[3,4-C]pyridine solution was prepared by dissolving 2,3-dihydro-1H-pyrrolo[3,4-C]pyridine (270 mg, 1.72 mmol) and DIPEA (446 mg, 3.45 mmol) in anhydrous acetonitrile / DMF (0.95 mL / 0.48 mL). After the two solutions were mixed, they were transferred to room temperature for reaction for 12 hours. After the reaction was complete, water was added for dilution, and ethyl acetate (15 mL × 3) was used for extraction. The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain intermediate 9 (450 mg, 52%). MS (ESI, m / z): 501 (M+ +1).

[0084] b) Intermediate 9 (450 mg, 0.90 mmol) was dissolved in DCM (2 mL), TFA (1 mL) was added to the reaction system, and the reaction was allowed to react at room temperature for 1 hour. After the reaction was complete, dichloromethane was removed by distillation under reduced pressure, and water was added to dilute the mixture. A 2M aqueous solution of NaOH was added to adjust the pH to 8-10 at 0°C, and the mixture was extracted with dichloromethane. The organic phases were combined, dried over anhydrous Na2SO4, filtered to remove salt, and concentrated under reduced pressure to obtain Intermediate 10 (295 mg, 82%). MS (ESI, m / z): 401 (M + +1).

[0085] Step 2: Synthesis of compound S3

[0086]

[0087] a) Intermediate 10 (75 mg, 0.19 mmol) and compound 8 (56 mg, 0.19 mmol) were dissolved in DMF (1 mL), and BOP (124 mg, 0.28 mmol) and DIPEA (73 mg, 0.56 mmol) were added to the reaction system. The mixture was reacted at room temperature for 12 hours. After the reaction was complete, the mixture was diluted with water and extracted with ethyl acetate (10 mL×3). The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain S3 (57 mg, 45%).

[0088] S3: 1 H NMR (400MHz, DMSO-d6) δ12.60(s,1H),8.59(s,1H),8.48(d,J=5.0Hz,1H),8.29-8.23(m,1H),7.97(d,J=7.9Hz,1H) ,7.91-7.75(m,4H),7.61(d,J=8.1Hz,2H),7.41(d,J=5.4Hz,2H),7.33(s,1H),7.27-7.13(m,2H),4.69(d,J=7.8Hz, 4H),4.55(d,J=13.3Hz,1H),4.42(d,J=5.8Hz,2H),4.32(s,2H),3.55(t,J=12.2Hz,1H),3.41(s,1H),3.01(t,J=12 .9Hz,1H),2.75(t,J=12.7Hz,1H),1.93(d,J=12.8Hz,1H),1.77(s,1H),1.41(d,J=24.3Hz,2H).MS(ESI,m / z):681(M + +1).

[0089] Example 4: Synthesis of Compound S4

[0090]

[0091] The synthesis method is similar to that of Example 1, and compound S4 can be obtained by simply replacing the corresponding raw materials.

[0092] S4: 1 H NMR (300MHz, DMSO-d6) δ12.59(s,1H),12.16(s,1H),9.42(t,J=6.1Hz,1H),8.80(s,1H),8.25(dd,J=7.3,1.7H z,1H),8.13(d,J=5.6Hz,1H),8.00-7.92(m,1H),7.92-7.78(m,4H),7.69-7.58(m,3H),7.46-7.35(m,1H),7.3 2(s,1H),7.27-7.15(m,2H),4.65(d,J=5.9Hz,2H),4.54(d,J=13.2Hz,1H),4.31(s,2H),3.55(s,1H),3.00(s, 1H),2.74(t,J=12.4Hz,1H),1.92(d,J=12.7Hz,1H),1.76(s,1H),1.37(d,J=15.1Hz,2H).MS(ESI,m / z):679(M + +1).

[0093] Example 5: Synthesis of Compound S5

[0094]

[0095] The synthesis method is similar to that of Example 1, and compound S5 can be prepared by simply replacing the corresponding raw materials.

[0096] S5: 1HNMR (300MHz, DMSO-d6) δ12.60(s,1H),12.16(s,1H),9.42(t,J=6.1Hz,1H),8.81(s,1H),8.24(dd,J=7.3,1.7 Hz,1H),8.13(d,J=5.6Hz,1H),7.99-7.90(m,1H),7.94-7.79(m,4H),7.67-7.56(m,3H),7.48-7.35(m,1H),7. 30(s,1H),7.29-7.14(m,2H),4.65(d,J=5.9Hz,2H),4.55(d,J=13.2Hz,1H),4.30(s,2H),3.56(s,1H),3.02(s ,1H),2.76(t,J=12.4Hz,1H),1.93(d,J=12.7Hz,1H),1.77(s,1H),1.39(d,J=15.1Hz,2H).MS(ESI,m / z):679(M + +1).

[0097] Example 6: Synthesis of Compound S6

[0098]

[0099] The synthesis method is similar to that of Example 1, and compound S6 can be prepared by simply replacing the corresponding raw materials.

[0100] S6: 1 H NMR (400MHz, DMSO-d6) δ12.60(s,1H),9.67(t,J=6.0Hz,1H),9.33(s,1H),8.54(d,J=5.4Hz,1H),8.26(d,J=7. 8Hz,1H),8.21(s,1H),7.96(t,J=6.0Hz,2H),7.93-7.77(m,4H),7.65(d,J=8.1Hz,2H),7.40(t,J=6.5Hz,1H),7 .32(s,1H),7.20(t,J=9.0Hz,1H),4.64(d,J=5.9Hz,2H),4.55(d,J=13.3Hz,1H),4.31(s,2H),3.57(s,1H),3. 01(s,1H),2.75(t,J=12.7Hz,1H),1.93(d,J=12.9Hz,1H),1.77(s,1H),1.56-1.28(m,2H).MS(ESI,m / z):696(M + +1).

[0101] Example 7: Synthesis of Compound S7

[0102]

[0103] The synthesis method is similar to that of Example 1, and compound S7 can be prepared by simply replacing the corresponding raw materials.

[0104] S7: 1 HNMR (400MHz, DMSO-d6) δ12.59(s,1H),9.70(t,J=6.0Hz,1H),9.38(s,1H),8.53(d,J=5.4Hz,1H),8.27(d,J=7 .8Hz,1H),8.24(s,1H),7.98(t,J=6.0Hz,2H),7.95-7.79(m,4H),7.67(d,J=8.1Hz,2H),7.43(t,J=6.5Hz,1H), 7.32(s,1H),7.22(t,J=9.0Hz,1H),4.67(d,J=5.9Hz,2H),4.55(d,J=13.3Hz,1H),4.32(s,2H),3.56(s,1H),3. 02(s,1H),2.74(t,J=12.7Hz,1H),1.93(d,J=12.9Hz,1H),1.76(s,1H),1.57-1.30(m,2H).MS(ESI,m / z):680(M + +1).

[0105] Example 8: Synthesis of Compound S8

[0106]

[0107] The synthesis method is similar to that of Example 1, and compound S8 can be prepared by simply replacing the corresponding raw materials.

[0108] S8: 1HNMR (400MHz, DMSO-d6) δ12.60(s,1H),9.37(t,J=6.0Hz,1H),9.06(d,J=2.1Hz,1H),8.79(d,J=2.1Hz,1H),8.32(s,1H),8.28 -8.20(m,2H),7.98(d,J=7.9Hz,1H),7.89-7.77(m,4H),7.64(d,J=8.0Hz,2H),7.45-7.38(m,1H),7.35(s,1H),7.21(t,J=9.0 Hz,1H),4.63(d,J=5.8Hz,2H),4.57(d,J=13.0Hz,1H),4.32(s,2H),3.58(t,J=23.1Hz,1H),3.01(t,J=26.7,12.5Hz,1H),3.0 1(d,J=25.7Hz,1H),2.75(t,J=12.5Hz,1H),1.93(d,J=12.6Hz,1H),1.78(s,1H),1.35(d,J=23.7Hz,2H).MS(ESI,m / z):680(M + +1).

[0109] Example 9: Synthesis of Compound S9

[0110]

[0111] Step 1: Synthesis of intermediate 15

[0112]

[0113] a) Compound 11 (201 mg, 1.0 mmol) was dissolved in anhydrous DCM (4 mL), 1-tert-butyloxycarbonylpiperazine (220 mg, 1.18 mmol) and TEA (304 mg, 3 mmol) were added to the reaction system, and the mixture was reacted at room temperature for 1 hour. After the reaction was complete, water was added to dilute the mixture, and the mixture was extracted with dichloromethane. The organic phases were combined, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain intermediate 12 (250 mg, 71%). MS (ESI, m / z): 352 (M + +1).

[0114] b) Intermediate 12 (1.0 g, 2.85 mmol) and nickel chloride hexahydrate (675 mg, 2.85 mmol) were dissolved in MeOH / THF (2 mL / 4 mL), sodium borohydride (161 mg, 4.27 mmol) was added in batches at 0°C, and the mixture was reacted at 60°C for 12 hours. After the reaction was complete, the mixture was filtered through diatomaceous earth, the filtrate was cooled to 0°C, 2M HCl aqueous solution was added to quench the sodium borohydride, and the mixture was extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain intermediate 13 (500 mg, 49%). MS (ESI, m / z): 356 (M + +1).

[0115] c) Intermediate 13 (500 mg, 1.41 mmol) was dissolved in DMF (4 mL), and imidazo[1,2-A]pyridine-7-carboxylic acid (229 mg, 1.41 mmol), BOP (935 mg, 2.12 mmol) and DIPEA (698 mg, 4.23 mmol) were added to the reaction system. The reaction was allowed to react at room temperature for 10 hours. After the reaction was complete, water was added to dilute the mixture, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain intermediate 14 (540 mg, 77%). MS (ESI, m / z): 500 (M + +1).

[0116] d) Intermediate 14 (540 mg, 1.08 mmol) was dissolved in DCM (6 mL), TFA (3 mL) was added to the reaction system, and the reaction was allowed to react at room temperature for 1 hour. After the reaction was complete, dichloromethane was removed by distillation under reduced pressure, and water was added to dilute the mixture. A 2M aqueous solution of NaOH was added to adjust the pH to 8-10 at 0°C, and the mixture was extracted with dichloromethane. The organic phases were combined, dried over anhydrous Na2SO4, filtered to remove salt, and concentrated under reduced pressure to obtain Intermediate 15 (410 mg, 95%). MS (ESI, m / z): 400 (M + +1).

[0117] Step 2: Synthesis of compound S9

[0118]

[0119] a) Intermediate 15 (617 mg, 1.29 mmol) and compound 8 (241 mg, 1.29 mmol) were dissolved in DMF (7 mL), and BOP (856 mg, 1.94 mmol) and DIPEA (44 mg, 0.34 mmol) were added to the reaction system. The mixture was reacted at room temperature for 12 hours. After the reaction was complete, the mixture was diluted with water and extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain S9 (550 mg, 52%).

[0120] S9: 1 H NMR (400MHz, DMSO-d6) δ12.57(s,1H),9.24(s,1H),9.18(s,H),8.26(d,J=7 .4Hz,1H),8.08(s,1H),7.94-7.81(m,3H),7.74-7.61(m,7H),7.40(s,1H),7 .29(d,J=5.8Hz,1H),7.18(t,J=9.0Hz,1H),4.62(d,J=5.8Hz,2H),4.29(s,2 H),3.69(s,2H),3.25(s,2H),2.96(s,2H),2.82(s,2H).MS(ESI,m / z):680(M + +1).

[0121] Example 10: Synthesis of Compound S10

[0122]

[0123] The synthesis method is similar to that of Example 3. Compound S10 can be prepared by simply replacing intermediate 5 with intermediate 13.

[0124] S10: 1 H NMR (400MHz, DMSO-d6) δ12.60 (s, 1H), 8.56 (s, 1H), 8.48 (d, J = 5.0Hz, 1H), 8.29-8.2 4(m,1H),7.96(d,J=7.9Hz,1H),7.90-7.74(m,4H),7.63(d,J=8.1Hz,2H),7.42(d,J =5.4Hz,2H),7.33(s,1H),7.28-7.12(m,2H),4.74(s,4H),4.63(d,J=6.8Hz,2H),4. 35(s,2H),3.77(s,2H),3.28(s,2H),3.11(s,2H),2.92(s,2H).MS(ESI,m / z):682(M + +1).

[0125] Example 11: Synthesis of Compound S11

[0126]

[0127] Step 1: Synthesis of intermediate 18

[0128]

[0129] a) Intermediate 16 (200 mg, 0.72 mmol), N,N'-disuccinimidyl carbonate (221 mg, 0.86 mmol) and pyridine (57 mg, 0.72 mmol) were dissolved in anhydrous acetonitrile (3.6 mL), and 2,3-dihydro-1H-pyrrolo[3,4-C]pyridine solution was added dropwise at 0°C. 2,3-dihydro-1H-pyrrolo[3,4-C]pyridine solution was prepared by dissolving 2,3-dihydro-1H-pyrrolo[3,4-C]pyridine (86 mg, 0.72 mmol) and DIPEA (186 mg, 1.44 mmol) in anhydrous acetonitrile / DMF (0.4 mL / 0.2 mL). After the two solutions were mixed, they were transferred to room temperature for reaction for 12 hours. After the reaction was complete, water was added for dilution, and ethyl acetate (10 mL × 3) was used for extraction. The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain intermediate 17 (152 mg, 50%). MS (ESI, m / z): 423 (M + +1).

[0130] b) Intermediate 17 (152 mg, 0.36 mmol) was dissolved in DCM (2 mL), TFA (1 mL) was added to the reaction system, and the reaction was allowed to react at room temperature for 1 hour. After the reaction was complete, dichloromethane was removed by distillation under reduced pressure, and water was added to dilute the mixture. A 2M aqueous solution of NaOH was added to adjust the pH to 8-10 at 0°C, and the mixture was extracted with dichloromethane. The organic phases were combined, dried over anhydrous Na2SO4, filtered to remove salt, and concentrated under reduced pressure to obtain Intermediate 18 (105 mg, 90%). MS (ESI, m / z): 323 (M + +1).

[0131] Step 2: Synthesis of compound S11

[0132]

[0133] a) Intermediate 21 (127 mg, 0.29 mmol) and compound 8 (54 mg, 0.29 mmol) were dissolved in DMF (1 mL), and BOP (194 mg, 0.44 mmol) and DIPEA (113 mg, 0.88 mmol) were added to the reaction system. The mixture was reacted at room temperature for 12 hours. After the reaction was complete, the mixture was diluted with water and extracted with ethyl acetate (10 mL×3). The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain S11 (109 mg, 46%).

[0134] S11: 1 H NMR(400MHz,DMSO-d6)δ12.60(s,1H),8.29-8.24(m,1H),8.16(s,1H),8.03-7.96(m,1H),7.93-7.81(m,2H),7.49-7.22(m, 9H),6.92-6.84(m,2H),4.74(s,4H),4.34(s,2H),3.76(s,2H),3.29(s,2H),3.11(s,2H),2.95(s,2H).MS(ESI,m / z):603(M + +1).

[0135] Example 12: Synthesis of Compound S12

[0136] Step 1: Synthesis of intermediate 21

[0137]

[0138] a) Intermediate 19 (476 mg, 1.72 mmol), N,N'-disuccinimidyl carbonate (530 mg, 2.07 mmol) and pyridine (136 mg, 1.72 mmol) were dissolved in anhydrous acetonitrile (8.6 mL), and 2,3-dihydro-1H-pyrrolo[3,4-C]pyridine solution was added dropwise at 0°C. 2,3-dihydro-1H-pyrrolo[3,4-C]pyridine solution was prepared by dissolving 2,3-dihydro-1H-pyrrolo[3,4-C]pyridine (270 mg, 1.72 mmol) and DIPEA (446 mg, 3.45 mmol) in anhydrous acetonitrile / DMF (0.95 mL / 0.48 mL). After the two solutions were mixed, they were transferred to room temperature for reaction for 12 hours. After the reaction was complete, water was added for dilution, and ethyl acetate (15 mL × 3) was used for extraction. The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain intermediate 20 (510 mg, 55%). MS (ESI, m / z): 423 (M + +1).

[0139] b) Intermediate 20 (510 mg, 1.2 mmol) was dissolved in DCM (2 mL), TFA (1 mL) was added to the reaction system, and the reaction was allowed to react at room temperature for 1 hour. After the reaction was complete, dichloromethane was removed by distillation under reduced pressure, water was added to dilute, 2M NaOH aqueous solution was added to adjust the pH to 8-10 at 0°C, and dichloromethane was used for extraction. The organic phases were combined, dried over anhydrous Na2SO4, filtered to remove salt, and concentrated under reduced pressure to obtain intermediate 21 (350 mg, 90%). MS (ESI, m / z): 323 (M + +1).

[0140] Step 2: Synthesis of compound S12

[0141]

[0142] a) Intermediate 21 (120 mg, 0.37 mmol) and compound 8 (90 mg, 0.30 mmol) were dissolved in DMF (1 mL), and BOP (200 mg, 0.45 mmol) and DIPEA (232 mg, 1.80 mmol) were added to the reaction system. The mixture was reacted at room temperature for 12 hours. After the reaction was complete, the mixture was diluted with water and extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed twice with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered to remove salt, concentrated under reduced pressure, and purified by column chromatography to obtain S12 (108 mg, 48%).

[0143] S12: 1 H NMR (300MHz, DMSO-d6) δ12.62(s,1H),8.61(s,1H),8.50(d,J=5.0Hz,1H),8.39(s,1H),8.26(dd,J=7.6,1.6 Hz,1H),7.98(d,J=7.8Hz,1H),7.93-7.77(m,2H),7.49(d,J=8.2Hz,2H),7.42(dd,J=8.9,5.1Hz,3H),7.24(t ,J=9.0Hz,1H),7.13(d,J=8.5Hz,2H),4.80(d,J=5.4Hz,4H),4.62(d,J=12.8Hz,1H),4.35(s,2H),3.43(s,1H ),3.12(t,J=12.6Hz,1H),2.90-2.67(m,2H),1.86(d,J=12.8Hz,1H),1.73-1.36(m,3H).MS(ESI,m / z):603(M + +1).

[0144] Example 13: Synthesis of Compound S13

[0145]

[0146] The synthesis method can be referred to Example 12, and compound S13 can be prepared by simply replacing the corresponding raw materials.

[0147] S13: 1 H NMR (400MHz, DMSO-d6) δ12.61(s,1H),8.64(d,J=2.6Hz,1H),8.54(s,1H),8.26(d, J=7.8Hz,1H),7.99(d,J=7.9Hz,1H),7.96-7.78(m,3H),7.45-7.29(m,6H),7.28-7. 15(m,2H),4.78(s,4H),4.60(d,J=12.9Hz,1H),4.35(s,2H),3.20-3.09(m,2H),2.8 9(d,J=12.1Hz,2H),1.93(d,J=13.0Hz,1H),1.80-1.44(m,3H).MS(ESI,m / z):603(M + +1).

[0148] Example 14: Synthesis of Compound S14

[0149]

[0150] The synthesis method can be referred to Example 12, and compound S14 can be prepared by simply replacing the corresponding raw materials.

[0151] S14: 1 H NMR (400MHz, DMSO-d6) δ12.61(s,1H),8.96(s,1H),8.25(d,J=7.8Hz,1H),8.15(d,J=2.3Hz,1H),7.97 (d,J=7.9Hz,1H),7.91-7.76(m,3H),7.65(d,J=8.8Hz,1H),7.46-7.39(m,2H),7.39-7.27(m,4H),7.23 (t,J=9.0Hz,1H),4.80(s,4H),4.63(d,J=12.9Hz,1H),4.34(s,2H),3.42(s,1H),3.13(t,J=12.7Hz,1H ),2.82(q,J=12.2Hz,2H),1.96-1.75(m,1H),1.58(d,J=12.4Hz,2H),1.45(s,1H).MS(ESI,m / z):603(M + +1).

[0152] Example 15: Synthesis of Compound S15

[0153]

[0154] The synthesis method can be referred to Example 11, and compound S15 can be prepared by simply replacing the corresponding raw materials.

[0155] S15: 1 H NMR(400MHz,DMSO-d6)δ12.61(s,1H),8.29-8.26(m,1H),8.15(s,1H),8.03-7.97(m,1H),7.93-7.83(m,2H),7.46-7.23(m,9H),6 .93-6.86(m,2H),4.73(s,4H),4.34(s,2H),3.72(s,2H),3.43(s,1H),3.29(s,2H),3.10(s,2H),2.96(s,2H).MS(ESI,m / z):602(M + +1).

[0156] Example 16: Evaluation of PARP and NAMPT enzyme inhibitory activity

[0157] 1. Experimental methods

[0158] The PARP1 enzyme inhibition activity was measured using the PARP homogeneous test kit. The test system included 1 ng / μL PARP-1, 20 ng / μL activated DNA (Sigma), and 500 nM NAD dissolved in the following reaction buffer: 100 mM Tris-HCl (pH 8.0), 100 mM NaCl, and 20 mM MgCl2. At room temperature, PARP-1 protein (1 ng / μL) was pre-incubated with different concentrations of the test compound for 30 min, and then 500 nM NAD was added to start the PARylation reaction. The remaining unreacted NAD was incubated with the circulating test solution (1% ethanol, 25 μM resazurin, 0.30 U / mL alcohol dehydrogenase, and 0.25 U / mL diaphorase), and the concentration of NAD was proportional to the fluorescence signal of Ex 540 nm / Em 590 nm. The reduction rate of NAD under the action of different concentrations of compounds was used to evaluate the inhibitory activity IC of the compound on PARP1. 50 The numerical value of .

[0159] The NAMPT inhibitory activity test was performed according to the operating procedures provided by the manufacturer of the CycLex NAMPT colorimetric assay kit (MBL International Corp.). The basic principle is: NAM and PRPP generate NMN under the action of NAMPT, and NMN and ATP generate NAD under the action of NMNAT1, NAD generates NADH under the action of alcohol dehydrogenase (ADH), and NADH returns to NAD under the action of diaphorase. WST-1 forms orange-yellow formazan in the NAD / NADH enzyme catalytic cycle, and the effect of the compound on the NAMPT enzyme activity can be detected by detecting the change in absorbance at OD 450nM.

[0160] The specific experimental steps are as follows:

[0161] (1) Preparation of test compound solutions: All compounds were dissolved in DMSO with a stock solution concentration of 10 mM. The required concentrations were prepared according to the test requirements. Starting from the starting concentration, 2-fold gradient dilution was performed to set 8 concentration gradients, and each concentration was repeated three times.

[0162] (2) ddH2O (10 μL) and NAM (5 μL) were added to a 96-well assay plate;

[0163] (3) Add 5 μL of the test compound or DMSO to a 96-well assay plate;

[0164] (4) Prepare mixture I, including: 20×NAMPT assay buffer (5 μL), PRPP (5 μL), ATP (5 μL), recombinant NMNAT1 (5 μL), ddH2O (35 μL) and NAMPT (5 μL), totaling 60 μL. Add mixture I to a 96-well assay plate;

[0165] (5) Place the 96-well assay plate in a 30°C incubator and incubate for 60 min;

[0166] (6) Prepare mixture I, including: WST-1 (5 μL), ADH (5 μL), diaphorase (5 μL), ethanol (5 μL), totaling 20 μL. After incubation, remove the 96-well assay plate and add mixture II to the 96-well assay plate;

[0167] (7) Dynamically monitor the absorbance of each well at OD 450 nm on a microplate reader within 30 minutes, once every 5 minutes;

[0168] (8) The time period in which the absorbance and time showed a linear change was selected to calculate the reaction rate. The data were processed using GraphPad Prism5 software. In order to compensate for the NAMPT inhibitory activity of DMSO, the values ​​were corrected using DMSO as a solvent control.

[0169] 2. Experimental results

[0170] Table 1 Inhibitory activity of NAMPT and PARP1 (IC 50 , nM)

[0171]

[0172] As shown in Table 1, the compounds designed in the present invention have good inhibitory activity against NAMPT and PARP1, and the enzyme inhibition IC 50 The concentration value reached nanomolar level.

[0173] Example 17: Evaluation of anti-tumor activity

[0174] 1. Experimental methods

[0175] (1) Collect cells in the logarithmic growth phase, adjust the concentration of the cell suspension, and add 100 μL of the cell suspension to each well of a 96-well plate; the number of cells in each well is about 7,000, and incubate overnight at 5% CO2 and 37°C until the cells are completely attached to the wall;

[0176] (2) Setting a concentration gradient of the compound of the present invention, with 3 replicate wells for each concentration gradient, diluting the compound of the present invention into the corresponding culture medium to the desired final concentration, aspirating the original culture medium in the 96-well plate, adding 100 μL of the prepared culture medium containing the drug at the desired final concentration, and incubating at 5% CO2 and 37°C; and setting a blank group (containing only 100 μL of culture medium without cells, and subsequent treatment is the same as that of other wells) and a control group (containing cells and culture medium);

[0177] (3) After 68 hours of drug treatment, 10 μL MTT solution (5 mg / ml) was added to each well and cultured for another 4 hours (cells were treated with drugs for a total of 72 hours);

[0178] (4) Aspirate the culture medium in the wells (if the cells are suspended, centrifuge at 2500 rpm for 5 min before aspirating the culture medium). Add 150 μL of dimethyl sulfoxide to each well and shake until the crystals are fully dissolved. Detect the absorbance of each well at OD490 nm on an ELISA reader;

[0179] (5) Calculate the inhibition rate: inhibition rate = 1 - (OD value of the drug-treated group - OD value of the blank group) / (OD value of the control group - OD value of the blank group) = (OD value of the control group - OD value of the drug-treated group) / (OD value of the control group - OD value of the blank group);

[0180] (6) IC was calculated using GraphPad Prism fitting 50 value.

[0181] 2. Experimental results

[0182] Table 2 Anti-cell proliferation activity (IC 50 , μM)

[0183] Compound No. MDA-MB-436 HCC1937 MDA-MB-468 MCF-7 MDA-MB-231 MCF-10A S1 0.11 0.23 0.50 0.42 1.82 ＞50 S2 0.22 0.18 0.29 0.53 0.99 ＞50 S3 0.10 0.12 0.25 0.19 1.07 ＞50 S4 0.33 0.21 0.54 0.88 2.33 ＞50 S5 0.22 0.18 0.21 0.18 0.65 ＞50 S7 0.18 0.11 0.32 0.039 0.056 ＞50 S8 0.32 0.22 0.22 0.45 0.46 ＞50 S12 0.043 0.033 0.034 0.012 0.028 ＞50 FK866 19.2 20.3 40.3 9.8 34.5 ＞50 Olaparib 0.32 0.12 3.89 10.21 7.95 ＞50

[0184] As shown in Table 2, the compounds designed by the present invention have good anti-proliferative activity against BRAC mutant triple-negative breast cancer cells (MDA-MB-436 and HCC1937) and BRAC wild-type triple-negative breast cancer cells (MDA-MB-468, MCF-7, MDA-MB-231); and have no cytotoxicity to normal cells MCF-10A, indicating that the compounds designed by the present invention have good safety.

Claims

1. A phthalazinone compound as shown in formula I or a pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, prodrug, solvate, deuterated product, hydrate thereof, In Formula I: X is N or C; L1 is a single bond or Y is C 6-10 Aryl or 5-10 membered heteroaryl containing 1-3 N, O, S heteroatoms; L2 is -(CH2) n -, n is 0, 1, 2 or 3; R1 is a 9- to 10-membered bicyclic heteroaryl group containing 1 N and 1 to 2 N, O, and S heteroatoms.

2. The phthalazinone compound or the pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, prodrug, solvate, deuterated substance or hydrate thereof according to claim 1, characterized in that: In Formula I: Y is phenyl or pyridyl; n is 0, 1 or 2.

3. The phthalazinone compound or pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, prodrug, solvate, deuterated substance or hydrate thereof according to claim 1, characterized in that: In Formula I: Y is phenyl, n is 0 or 1.

4. The phthalazinone compound or a pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, prodrug, solvate, deuterated substance or hydrate thereof according to claim 1, characterized in that: In Formula I: R1 is imidazopyridyl, imidazopyrimidinyl, pyrazolopyridyl, thienopyridinyl, furopyridinyl, tetrahydropyrrolopyridinyl or tetrahydropyrrolophenyl.

5. The phthalazinone compound or its pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, prodrug, solvate, deuterated substance, hydrate according to claim 1, characterized in that: In Formula I: R1 is 6. The phthalazinone compound or pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, prodrug, solvate, deuterated substance or hydrate thereof according to claim 1, characterized in that: Any one of the following compounds:

7. A method for preparing the phthalazinone compound or its pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, prodrug, solvate, deuterated product or hydrate according to claim 1, characterized in that: The following steps are involved: The compound of formula II and the compound of formula III undergo condensation reaction under the action of a base and a condensing agent to obtain the compound of formula I: Among them, X, Y, L1, L2, and R1 are as described in claim 1.

8. A pharmaceutical composition, characterized in that Containing a therapeutically effective amount of the phthalazinone compound according to claim 1 or a pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformational isomer, metabolite, prodrug, solvate, deuterated substance, hydrate thereof and a pharmaceutically acceptable carrier.

9. Use of the phthalazinone compound according to claim 1 or its pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformer, metabolite, prodrug, solvate, deuterated substance, hydrate or the pharmaceutical composition according to claim 8 in the preparation of a drug for a poly(ADP-ribose) polymerase inhibitor and / or a drug for a nicotinamide phosphoribosyltransferase inhibitor.

10. Use of the phthalazinone compound according to claim 1 or its pharmaceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer, tautomer, atropisomer, conformer, metabolite, prodrug, solvate, deuterated substance, hydrate or the pharmaceutical composition according to claim 8 in the preparation of a drug for preventing and / or treating cancer.

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