PARP inhibitors - quinoline carboxylic acid coupled derivatives and methods of making and use thereof

Abstract

This invention belongs to the field of antitumor drug technology, specifically relating to PARP inhibitor-quinoline carboxylic acid coupled derivative compounds, their preparation methods, and uses. This invention synthesizes a novel polyADP-ribose polymerase (PARP) inhibitor derivative, the structure of which includes: a first structural unit with the clinical PARP inhibitor olaparib as the core backbone, a linker arm L, and a quinoline carboxylic acid fragment R; wherein the structural types and substituents of the linker arm L and the quinoline carboxylic acid fragment R are adjustable. This invention also provides a method for preparing the compound, which is characterized by simple steps, strong versatility, and ease of structural expansion, suitable for constructing compound libraries with different combinations of linker arms and different quinoline carboxylic acid fragments. The compound can be used to prepare drugs or drug compositions for inhibiting tumor cell proliferation and treating tumor-related diseases, showing excellent application prospects.

Description

Technical Field

[0001] This invention belongs to the field of antitumor drug technology, specifically relating to PARP inhibitor-quinoline carboxylic acid coupling derivatives, their preparation methods, and uses. Background Technology

[0002] In recent years, the DNA Damage Response (DDR) pathway has been considered one of the most promising target systems in cancer therapy. Tumor cells, during rapid proliferation, are highly dependent on the DNA repair system to maintain genomic stability, with PARP family proteins playing a central role in recognizing DNA single-strand breaks, recruiting repair proteins, and initiating the base excision repair (BER) pathway. Poly ADP-ribose polymerase (PARP) is a class of intranuclear enzymes that play a crucial role in DNA single-strand break repair, with PARP1 and PARP2 being particularly important in the BER pathway. When PARP function is inhibited, DNA single-strand breaks are converted into double-strand breaks, thereby inducing a "synthetic lethality" effect in tumor cells deficient in homologous recombination repair (HR). Olaparib, as the first approved PARP inhibitor, has been widely used in the treatment of BRCA1 / 2 mutation-related ovarian, breast, prostate, and pancreatic cancers. However, as clinical application has deepened, olaparib has gradually revealed a series of limitations, including: (1) after long-term use, tumor cells develop drug resistance by restoring HR repair or upregulating drug efflux pumps; (2) the drug molecules are highly polar, and there is still room for further optimization in tissue distribution and pharmacokinetic behavior; (3) the single-target inhibition mode has limited therapeutic depth in the complex tumor microenvironment.

[0003] Quinoline and its carboxylic acid derivatives are a class of heterocyclic structural units with a broad research foundation in areas such as antitumor, antibacterial, and kinase inhibition. Previous studies have shown that the quinoline backbone can interact with DNA, topoisomerases, or various protein kinases, and can be introduced as a functional module to achieve multi-target regulation or synergistic pharmacological effects. However, whether quinoline or its quinoline carboxylic acid derivatives can enhance the antitumor activity of PARP inhibitors remains unknown.

[0004] Therefore, constructing a conjugate of a PARP inhibitor with quinoline or its quinoline carboxylic acid derivative in order to improve the antitumor activity and drug resistance of existing PARP inhibitors is a problem facing this field. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention proposes a novel molecular design strategy, providing PARP inhibitor-quinoline carboxylic acid conjugated derivatives, their preparation methods, and applications. The aim is to provide a class of novel PARP inhibitor derivatives with the poly-ADP-ribose polymerase (PARP) inhibitor olaparib as the core pharmacodynamic framework and incorporating quinoline carboxylic acid structural units. These derivatives are intended to enhance the effects of DNA damage accumulation and anti-tumor cell proliferation inhibition, and to provide a new structural basis for further optimizing the anti-tumor activity and drug resistance-related studies of existing PARP inhibitors at the cellular level. To achieve the above objectives, this invention provides the following technical solutions: This invention provides a PARP inhibitor-quinoline carboxylic acid coupling derivative compound, or its optical isomer, or its tautomer, or its deuterated compound, or its tritated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form, wherein the structure of the PARP inhibitor-quinoline carboxylic acid coupling derivative compound is shown in Formula I: in, L is selected from -NH-L x -NH-、 ; L x Selected from C 2-4 Alkylene; m is 1 or 2; R is -C(O)R1; R1 is selected from the substituted or unsubstituted quinoline ring of R2, and R2 is selected from halogen, -OH, C 1-6 Alkyl, C 1-6 Haloalkyl, C 6-15 At least one of the aromatic rings.

[0006] Preferably, L is selected from -NH-L x -NH-、 ; L x Selected from C 2-4 Alkylene; m is 1 or 2; R is -C(O)R1; R1 is selected from a quinoline ring that is substituted or unsubstituted by R2, and R2 is selected from at least one of F, Cl, -OH, methyl, benzene ring, and trifluoromethyl.

[0007] Preferably, the number of R2 is 0, 1 or 2; The linker site of R1 to the carbonyl group is located at position 2, 4, 6, 7 or 8 of the quinoline ring; The connection site between R1 and R2 is located at at least one of the positions 2, 3, 6, 7, and 8 of the quinoline ring.

[0008] Preferred, L is selected from , , , One of them; L is In this case, R1 is selected from the quinoline ring with or without R2 substituted or unsubstituted, and the linking site of R1 to the carbonyl group is located at position 2, 4, 6, 7 or 8 of the quinoline ring. R2 is selected from phenyl, methyl, Cl, F, trifluoromethyl, and the number of R2 is 0, 1 or 2. The linking site of R1 and R2 is located at at least one of the positions 2, 3, 6, 7 or 8 of the quinoline ring. L is or In this case, R1 is selected from the quinoline ring substituted with R2, the linking site of R1 to the carbonyl group is located at the 6 or 8 position of the quinoline ring, R2 is at least one of methyl and Cl, the number of R2 is 0 or 2, and the linking site of R1 to R2 is located at at least one of the 3 or 7 positions of the quinoline ring. L is In this case, R1 is selected from the quinoline ring that is substituted or unsubstituted with R2, the linking site of R1 to the carbonyl group is located at the 6th or 8th position of the quinoline ring, R2 is at least one of methyl and Cl, the number of R2 is 0 or 2, and the linking site of R1 to R2 is located at at least one of the 3rd or 7th position of the quinoline ring.

[0009] Preferably, L is R1 is selected from the quinoline ring substituted with R2, and the linking site of R1 and the carbonyl group is located at the 4 position of the quinoline ring. R2 is selected from phenyl or trifluoromethyl, and the linking site of R1 and R2 is located at the 2 position of the quinoline ring.

[0010] Preferably, R is selected from one of the following structures: .

[0011] Preferably, the structural formula of the PARP inhibitor-quinoline carboxylic acid coupling derivative compound is shown in any of the following: .

[0012] This invention provides a method for preparing the PARP inhibitor-quinoline carboxylic acid conjugate compound, or its optical isomer, or its tautomer, or its deuterated compound, or its tritated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form as described in any of the preceding claims, comprising: ; Compound a reacts with ROH via a condensation reaction to produce the compound shown in Formula I.

[0013] Preferably, the condensation reaction is carried out under the action of a condensing agent and a base, wherein the condensing agent is selected from HATU, HBTU, DCC, EDCI or CDI, the base is selected from diisopropylethylamine, triethylamine or DMAP, and the solvent for the condensation reaction is selected from dichloromethane, N,N-dimethylformamide, tetrahydrofuran or acetonitrile. And / or, compound a is prepared by the following method: Step 1: 2-Formylbenzoic acid is mixed with dimethyl phosphite and reacted under the action of an alkoxide base to obtain compound 1; Step 2: Compound 1 is mixed with an organic base and 2-fluoro-5-formylbenzonitrile, and the reaction yields compound 2; Step 3: Compound 2 undergoes an alkaline conversion reaction, and then hydrazine hydrate is added to continue the reaction to obtain compound 3; Step 4: Compound 3 undergoes a condensation reaction with R3LH to obtain compound b, where R3 is an amino protecting group; Step 5: Compound b undergoes a deprotection reaction to obtain compound a.

[0014] This invention provides the use of any of the preceding PARP inhibitors—quinoline carboxylic acid conjugates, or their optical isomers, tautomers, deuterated compounds, tritated compounds, pharmaceutically acceptable salts, prodrugs, hydrates, solvates, or crystal forms—in the preparation of anti-breast cancer drugs.

[0015] The compounds and derivatives provided in this invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature systems.

[0016] Regarding the definition of terms used in this invention: Unless otherwise stated, the initial definitions provided for groups or terms herein apply to the groups or terms used throughout this specification; for terms not specifically defined herein, the meanings that a person skilled in the art would give them should be given based on the disclosure and context.

[0017] "Substitution" refers to the replacement of hydrogen atoms in a molecule by other different atoms or molecules.

[0018] The minimum and maximum carbon atom content in hydrocarbon groups are indicated by a prefix, for example, the prefix C. a-b Alkyl indicates any alkyl group containing "a" to "b" carbon atoms. Therefore, for example, "C 1-6 "Alkyl" refers to an alkyl group containing 1 to 6 carbon atoms.

[0019] "Alkyl" refers to a saturated hydrocarbon chain with a specified number of member atoms. For example, C 1-6 An alkyl group is an alkyl group having 1 to 6 member atoms, for example, 1 to 4 member atoms. An alkyl group can be straight-chain or branched. A representative branched alkyl group has one, two, or three branches. An alkyl group may optionally be substituted by one or more substituents as defined herein. Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and tert-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl. An alkyl group may also be part of other groups, such as C1-C6 alkoxy groups.

[0020] "Halogenated alkyl" refers to an alkyl group in which at least one hydrogen atom is replaced by a halogen atom.

[0021] "Cycloalkyl" refers to a saturated or partially saturated cyclic group having 3 to 14 carbon atoms, no heterocyclic atoms, and a single or multiple rings (including fused, bridged, and spirocyclic systems); wherein, when all chemical bonds in a cycloalkyl group are saturated, the cycloalkyl group is called a "saturated cycloalkyl"; when some chemical bonds in a cycloalkyl group are unsaturated, the cycloalkyl group is called a "partially unsaturated cycloalkyl". For polycyclic systems having aromatic and non-aromatic rings without heteroatoms, the term "cycloalkyl" (e.g., 5,6,7,8,-tetrahydronaphthyl-5-yl) applies when the bonding point is located on a non-aromatic carbon atom. The term "cycloalkyl" includes cycloalkenyl groups, such as cyclohexenyl. Examples of cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclooctyl, cyclopentenyl, and cyclohexenyl. Examples of cycloalkyl groups including polycycloalkyl ring systems are dicyclohexyl, dicyclopentyl, dicyclooctyl, etc.

[0022] "Partially unsaturated" means that it has both single, double, and triple bonds, and does not have aromaticity.

[0023] "Heterocycle" refers to a saturated ring or a non-aromatic unsaturated ring containing at least one heteroatom and having a single ring or multiple rings (including fused, bridged, and spirocyclic systems); where heteroatoms refer to nitrogen, oxygen, and sulfur atoms; "Aromatic ring" refers to an aromatic unsaturated ring that does not contain heteroatoms and has a single ring or multiple rings (including fused, bridged and spirocyclic systems).

[0024] "Heteroaromatic ring" or "heteroaryl" refers to an aromatic unsaturated ring containing at least one heteroatom and having a single or multiple rings; where heteroatoms refer to nitrogen, oxygen, or sulfur atoms.

[0025] The quinoline ring is a heteroaromatic ring. The numbering rule for the quinoline ring is as follows: the nitrogen atom in the quinoline ring is at position 1. Starting from N, the ring is numbered sequentially along the pyridine ring until the bridgehead carbon 4a. From 4a, the ring is numbered sequentially along the benzene ring until the other bridgehead carbon 8a is reached. See the details below: .

[0026] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0027] "Alkoxy" refers to a group in which an alkyl group is attached to a linker site via an oxygen atom. For example, methoxy is -OCH3.

[0028] "L1 and L2 are connected to form a ring" means that at least one atom in L1 and L2 are connected by a chemical bond, so that L1, L2 and the molecular skeleton they belong to together form a ring structure.

[0029] "Alkoxide base" refers to a compound formed when the hydrogen atom of the hydroxyl group in an alcohol molecule is replaced by a metal (usually an alkali metal such as sodium, potassium, or an alkaline earth metal); for example, sodium methoxide or sodium ethoxide.

[0030] The term "pharmaceutically acceptable" means that a carrier, delivery substance, diluent, excipient, and / or the salt formed therefrom is generally chemically or physically compatible with other components constituting a drug dosage form and physiologically compatible with receptors.

[0031] The terms "salt" and "pharmaceutical salt" refer to acidic and / or basic salts formed by the above-described compounds or their stereoisomers with inorganic and / or organic acids and bases, including zwitterionic salts (internal salts) and quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly during the final separation and purification of the compounds. Alternatively, they can be obtained by mixing the above-described compounds or their stereoisomers with an appropriate amount (e.g., equimolar amounts) of an acid or base. These salts may be obtained by precipitating in solution and collecting by filtration, by recovery after solvent evaporation, or by freeze-drying after reaction in an aqueous medium. The salts described in this invention can be hydrochlorides, sulfates, citrates, benzenesulfonates, hydrobromides, hydrofluoric acids, phosphates, acetates, propionates, succinates, oxalates, malates, succinates, fumarates, maleates, tartrates, or trifluoroacetates of the compounds.

[0032] In some embodiments, one or more compounds of the present invention may be used in combination with each other. Alternatively, the compounds of the present invention may be used in combination with any other active agent to prepare a medicament or pharmaceutical composition for regulating cell function or treating disease. If a group of compounds is used, these compounds may be administered to the test subject simultaneously, separately, or sequentially.

[0033] This invention provides a class of olaparib-quinoline carboxylic acid coupling derivatives, which consist of three parts: (1) PARP inhibitory core pharmacophore unit: olaparib or its coupling derivative backbone as the core pharmacophore structural unit. (2) Linker group (L): located between the olaparib core structural unit and the quinoline carboxylic acid fragment, used to achieve covalent connection, regulate spatial conformation, physicochemical properties and cell permeability. (3) Quinoline carboxylic acid structural unit (R): containing the quinoline ring and its carboxylic acid / carboxamide derivative structure, used to expand the molecular interaction space and provide further structural optimization sites. The general structural formula of the derivative is as follows (Formula (I)): Equation (I) Wherein: L is the linker arm; R is the quinoline carboxylic acid fragment, and the carboxylic acid group of R is connected to the linker arm L via an amide bond.

[0034] In this invention, the linker arm L connects the olaparib core skeleton to the quinoline carboxylic acid structural unit. By selecting the type and chain length of the linker arm L, the spatial conformation, physicochemical properties, and cell permeability of the compound can be controlled. In conjunction with the synthetic route described in this invention, the linker arm L is a diamine-type linker arm with amino groups at both ends, or a amino group that can be converted to amino functional groups. One amino group is used for covalent bonding with the olaparib core skeleton, and the other amino group is used to form an amide bond with the quinoline carboxylic acid structural unit. The linker arm L can be a commonly used diamine-type linker group in the art, including the following types: (1) Aliphatic diamine linker: a straight-chain or branched diamine segment, such as ethylenediamine, propylenediamine, butanediamine, pentanediamine, hexanediamine and their substituted derivatives; (2) Flexible diamine linker containing heteroatoms: a diamine segment containing O, N or S atoms, such as an amino ether or amino alkoxy linker; (3) Nitrogen-containing heterocyclic diamine linkers: for example, piperazine, piperidine, morpholine and their substituted derivatives; (4) Aromatic ring-containing diamine-type linker: for example, phenylene diamine or its substituted derivatives.

[0035] In a preferred embodiment, the connecting arm L is a piperazine or aliphatic diamine type connecting arm. In a preferred embodiment, the connecting arm L is a piperazine type connecting arm.

[0036] In one embodiment of the present invention, the quinoline carboxylic acid structural unit R is a quinoline carboxylic acid core formed by introducing carboxylic acid functional groups into the quinoline ring system and its substituted derivatives. The quinoline carboxylic acid core includes positional isomers formed by introducing carboxylic acid groups at different sites, and the core can be further substituted by substituents to achieve structural diversification and physicochemical property regulation. The quinoline carboxylic acid core includes, but is not limited to, quinoline-2-carboxylic acid, quinoline-3-carboxylic acid, quinoline-4-carboxylic acid, quinoline-5-carboxylic acid, quinoline-6-carboxylic acid, quinoline-7-carboxylic acid, and quinoline-8-carboxylic acid.

[0037] In a further embodiment, one or more substituents may be introduced onto the quinoline ring of the quinoline carboxylic acid core. The substitution sites of these substituents may be located at positions 5, 6, 7, or 8 of the quinoline ring (and other substitutable sites). The substituents are selected from: halogens, C... 1-6 Alkyl, C 1-6 Alkoxy, amino, hydroxy, nitro, cyano, trifluoromethyl, or combinations thereof.

[0038] In a preferred embodiment, the quinoline carboxylic acid structural unit R is any of the above-mentioned positioning isomers, and one or more of the above-mentioned substituents are introduced onto the quinoline ring to obtain a substituted quinoline carboxylic acid derivative.

[0039] This invention synthesizes a novel poly(ADP-ribose) polymerase (PARP) inhibitor derivative. This derivative uses the clinical PARP inhibitor olaparib as its core pharmacophore and is constructed by covalent coupling with a quinoline carboxylic acid derivative. The derivative is a compound of formula (I) and its pharmaceutically acceptable salt, solvate, or hydrate, and its structure includes: a first structural unit with olaparib as the core skeleton, a linker arm L, and a quinoline carboxylic acid fragment R; wherein the structural types and substituents of the linker arm L and the quinoline carboxylic acid fragment R are adjustable. This invention also provides a method for preparing the compound, which is simple, versatile, and facilitates structural expansion, suitable for constructing compound libraries with different combinations of linker arms and quinoline carboxylic acid fragments. The compound can be used to prepare drugs or pharmaceutical compositions for inhibiting tumor cell proliferation and treating tumor-related diseases.

[0040] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0041] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0042] Figure 1 This image shows the results of fluorescence microscopy analysis of tumor cell proliferation after treatment with compound I-9; in which, Figure 1 Figure A shows the effect of different concentrations of compound I-9 on the proliferation of 4T1 cells. Figure 1 Figure B shows the effect of different concentrations of compound I-9 on the proliferation of MCF-7 cells; DAPI represents the DAPI signal under a fluorescence microscope, EdU represents the EdU signal under a fluorescence microscope, and DAPI / EdU represents the mixed signal of DAPI and EdU under a fluorescence microscope. Detailed Implementation

[0043] Unless otherwise specified, all reagents and materials used in the following examples and experimental cases are commercially available.

[0044] In this invention, the preparation of olaparib-quinoline carboxylic acid derivatives employs a modular design and a general synthetic strategy. The overall synthetic route includes three main steps: obtaining the core pharmacophore structural unit of olaparib, constructing an olaparib-conjugable intermediate, and introducing the quinoline carboxylic acid structural unit. The synthetic route of olaparib-quinoline carboxylic acid derivatives is shown below: As shown in the synthetic route above, the core pharmacodynamic skeleton of olaparib (compounds 1-3) is first constructed through a multi-step reaction. Then, a linker arm L is introduced into the core skeleton to obtain an olaparib intermediate with couplerable functional groups (compound 4). Further, a condensation reaction is carried out with a quinoline carboxylic acid structural unit R to obtain the olaparib-quinoline carboxylic acid derivative (compound I) described in this invention. Here, L is the linker arm, the specific definition of which is given in the technical solution section of the specification; R is a quinoline carboxylic acid structural unit, which includes quinoline carboxylic acids at different sites and their substituted derivatives. By changing the structure of the linker arm L and the type of the quinoline carboxylic acid structural unit R, a series of structurally diverse olaparib-quinoline carboxylic acid derivatives can be obtained.

[0045] L contains the following structural formula: .

[0046] R contains the following structural formulas: .

[0047] Includes the following steps: (1) 2-Formylbenzoic acid and dimethyl phosphite are dissolved in an organic solvent and reacted at room temperature under the action of alkali to give dimethyl (3-oxo-1,3-dihydroisobenzofuran-1-yl)phosphonate (compound 1).

[0048] The organic solvent mentioned above can be methanol, ethanol or other lower alcohol solvents, preferably methanol; the base can be sodium methoxide or sodium ethoxide, preferably sodium methoxide.

[0049] (2) Compound 1 was dissolved in an organic solvent, and 2-fluoro-5-formylbenzonitrile was added under low temperature conditions in the presence of an organic base. The reaction was then continued at room temperature to obtain 2-fluoro-5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)benzonitrile (compound 2).

[0050] The organic solvent may be tetrahydrofuran, dichloromethane, acetonitrile, 1,4-dioxane, or toluene, preferably tetrahydrofuran; the base may be triethylamine, diisopropylethylamine, or potassium carbonate, preferably triethylamine.

[0051] (3) Compound 2 was added to an alkaline system and subjected to an alkaline conversion reaction at 90 °C. After the reaction was completed, hydrazine hydrate was added and the reaction was continued at 70 °C to obtain 2-fluoro-4-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoic acid (compound 3).

[0052] The base mentioned above can be sodium hydroxide or potassium hydroxide, preferably sodium hydroxide; the reaction solvent can be water, an alcohol / water mixture, or a tetrahydrofuran / water mixture, wherein the alcohol can be methanol or ethanol; in a preferred embodiment, the solvent is water or a methanol / water mixture.

[0053] (4) Compound 3 is dissolved in an organic solvent. Under the action of a condensing agent and a base, the carboxyl group of compound 3 reacts with a diamine-type linker with a protected amino group to form an amide bond, yielding an intermediate containing a Boc protecting group. Subsequently, the obtained intermediate is subjected to a deprotection reaction under acidic conditions to remove the Boc protecting group, yielding intermediate compound 4 with a free amino group at the end. In a preferred embodiment, the diamine-type linker with the protected amino group is N-Boc-piperazine.

[0054] The organic solvent mentioned above can be dichloromethane, N,N-dimethylformamide, tetrahydrofuran, or acetonitrile, preferably dichloromethane; the condensing agent can be HATU, HBTU, DCC, EDCI, or CDI, preferably HATU; the base can be diisopropylethylamine, triethylamine, or DMAP, preferably diisopropylethylamine; the acidic reagent for the deprotection reaction can be an organic solution of trifluoroacetic acid or hydrochloric acid, and the organic solvent for the deprotection reaction can be dichloromethane, chloroform, or tetrahydrofuran.

[0055] (5) Dissolve intermediate compound 4 in an organic solvent, and react with quinoline carboxylic acid structural units under the action of condensing agent and base to form amide bonds, thereby obtaining target compound I.

[0056] The organic solvent mentioned above can be dichloromethane, N,N-dimethylformamide, tetrahydrofuran, or acetonitrile, preferably dichloromethane or N,N-dimethylformamide; the condensing agent can be HATU, HBTU, DCC, EDCI, or CDI, preferably HATU or DCC; the base can be diisopropylethylamine, triethylamine, or DMAP, preferably diisopropylethylamine. The specific structure and substitution method of the quinoline carboxylic acid structural unit are detailed in the technical solution section of the specification.

[0057] Example 1 Synthesis of Compound I-1 Step (1): Synthesis of dimethyl (3-oxo-1,3-dihydroisobenzofuran-1-yl)phosphonate (compound 1) Sodium methoxide (3.24 g, 0.06 mol) was dissolved in anhydrous methanol (60.00 mL), and dimethyl phosphite (7.80 g, 0.06 mol) was slowly added at 0 °C. After stirring until homogeneous, 2-formylbenzoic acid (10.00 g, 0.06 mol) was added dropwise, and the mixture was heated to room temperature and stirred at this temperature for 8 h. Methanesulfonic acid was added dropwise to the reaction system, and the reaction was continued at room temperature for another half hour. After the reaction was detected by thin-layer chromatography (TLC), the mixture was cooled to room temperature, the pH of the reaction solution was adjusted to neutral with dilute hydrochloric acid, and the solvent was removed by vacuum distillation. The residue was diluted with water and extracted with dichloromethane (200 mL × 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate volume ratio = 2:1) to give compound 1 as a white or off-white solid. (12.10 g, yield: 75.2%). HRMS (m / z): [M+H] + calcd for C 10 H 12 O5P: 243.0417, found: 243.0399.

[0058] Step (2): Synthesis of 2-fluoro-5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)benzonitrile (compound 2) Compound 1 (5.00 g, 0.021 mol) was dissolved in tetrahydrofuran (20.00 mL). Triethylamine (4.26 g, 0.042 mol) was added under ice-water bath conditions. After stirring until homogeneous, 2-fluoro-5-formylbenzonitrile (3.36 g, 0.023 mol) was slowly added dropwise. After the addition was complete, the ice-water bath was removed, and the mixture was allowed to warm naturally to room temperature. The reaction was stirred for 18 h. After the reaction was monitored and completed, water was added to quench the reaction. The mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography with petroleum ether / ethyl acetate (volume ratio = 3:1) as the eluent to give compound 2 as a pale yellow solid (4.35 g, yield: 78.5%). HRMS (m / z): [M+H] + calcd for C 16 H9FNO2266.0612; found: 266.0602.

[0059] Step (3): Synthesis of 2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoic acid (compound 3) Compound 2 (3.00 g, 0.011 mol) was weighed and added to water (40.00 mL), followed by sodium hydroxide (1.32 g, 0.033 mol). The mixture was stirred at 90 °C for 1 h. After the reaction was confirmed by thin-layer chromatography, the mixture was cooled to room temperature, and hydrazine hydrate (NH₂NH₂·H₂O, 1.65 g, 0.033 mol) was added. The reaction mixture was then heated to 70 °C and stirred for another 18 h. After the reaction was completed, the mixture was cooled to room temperature, and the pH of the reaction solution was adjusted to acidic with dilute hydrochloric acid. A solid precipitated, which was collected by filtration, washed with water, and dried to give compound 3 as a white solid (2.36 g, yield: 72.1%). HRMS (m / z): [M+H] + calcd for C 16 H 12 FN2O3299.0821; found: 299.0816.

[0060] Step (4): Synthesis of tert-butyl 4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazine-1-carboxylate (compound 4) Compound 3 (2.00 g, 0.0067 mol) was dissolved in dichloromethane (40.00 mL). Under ice-water bath conditions, HATU (3.06 g, 0.0080 mol) and N,N-diisopropylethylamine (DIPEA, 2.60 g, 0.020 mol) were added sequentially. After stirring and activation for 30 min, N-Boc-piperazine (1.50 g, 0.0080 mol) was added. The ice-water bath was removed, and the mixture was allowed to warm naturally to room temperature while stirring for 2 h. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using petroleum ether / ethyl acetate (volume ratio = 1:1) to give compound 4 as a white or off-white solid (2.45 g, yield: 78.8%). HRMS (m / z): [M+H] + calcd for C 25 H 28 FN4O4467.2089; found: 467.2082.

[0061] Step (5): Synthesis of 4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazine-1(2H)-one (compound 8) Compound 4 (2.00 g, 0.0043 mol) was dissolved in dichloromethane (10.00 mL), and trifluoroacetic acid (TFA, 2.00 mL) was slowly added dropwise at room temperature with stirring for 1 h. After the reaction was detected by thin-layer chromatography (TLC), the solvent was removed under reduced pressure. The residue was dissolved in dichloromethane, and the solution was slowly added to a saturated sodium bicarbonate solution under ice-water bath conditions to neutralize to a weakly alkaline state. The solution was extracted with dichloromethane (100 mL × 3), and the organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography with dichloromethane / methanol (v / v = 20:1) as the eluent to give compound 8 as a white or off-white solid (1.47 g, yield: 93.6%). HRMS (m / z): [M+H] + calcd for C 20 H 20 FN4O2367.1565; found: 367.1554.

[0062] Step (6): Synthesis of 4-(4-fluoro-3-(4-(quinoline-2-carbonyl)piperazine-1-carbonyl)benzyl)phthalazine-1(2H)-one (compound I-1) Compound 8 (50.0 mg, 0.136 mmol) was weighed and dissolved in dichloromethane (3.00 mL). Quinoline-2-carboxylic acid (26.0 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 h. After the reaction was detected by thin-layer chromatography (TLC), water (20.0 mL) was added to quench the reaction. The mixture was separated, and the aqueous phase was extracted with dichloromethane (20.0 mL × 3). The combined organic phases were washed sequentially with saturated sodium bicarbonate aqueous solution (20.0 mL × 1) and saturated sodium chloride aqueous solution (20.0 mL × 1). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 30:1 → 20:1) as the eluent to give the target compound I-1 as a white or off-white solid (53.10 mg, yield: 75.3%). HRMS (m / z): [M+H] + calcd for C 30 H 25FN5O3522.1936;found: 522.1928.

[0063] 1 H NMR (600 MHz, DMSO- d 6) δ 12.57 (d, J = 27.9 Hz, 1H), 8.49 (d, J = 2.0Hz, 1H), 8.28 (s, 1H), 8.02 (d, J = 1.4 Hz, 2H), 7.88 (s, 2H), 7.84-7.75 (m,2H), 7.70-7.64 (m, 2H), 7.47-7.33 (m, 2H), 7.20 (d, J = 40.5 Hz, 1H), 4.33 (s,1H), 4.27 (s, 1H), 3.77 (s, 2H), 3.69 – 3.61 (m, 2H), 3.55 (s, 1H), 3.41 (s,1H), 3.20 (s, 1H), 2.78 (d, J = 94.2 Hz, 1H). Example 2 Synthesis of 4-(4-fluoro-3-(4-(quinoline-6-carbonyl)piperazine-1-carbonyl)benzyl)phthalazine-1(2H)-one (compound I-2) Compound 8 was synthesized according to the method of Example 1.

[0064] Compound 8 (50.0 mg, 0.136 mmol) was dissolved in dichloromethane (3.00 mL). Quinoline-6-carboxylic acid (26.0 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially under stirring, and the reaction was carried out at room temperature for 2 h. After the reaction was complete, water (20.0 mL) was added to quench the reaction mixture. The mixture was separated, and the aqueous phase was extracted with dichloromethane (20.0 mL × 3). The combined organic phases were washed sequentially with saturated sodium bicarbonate aqueous solution (20.0 mL × 1) and saturated sodium chloride aqueous solution (20.0 mL × 1). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 30:1 → 20:1) as the eluent to give the target compound I-2 as a white or off-white solid (56.30 mg, yield: 78.1%). HRMS (m / z): [M+H] + calcd for C 30 H 25 FN5O3522.1936; found: 522.1931.

[0065] 1 H NMR (600 MHz, DMSO- d 6) δ 12.60 (s, 1H), 8.97 (d, J = 4.2 Hz, 1H), 8.44 (d, J = 8.3 Hz, 1H), 8.30 (s, 1H), 8.08 (s, 2H), 7.81 (dd, J = 34.0, 8.2 Hz, 4H), 7.60 (dd, J = 8.3, 4.2 Hz, 1H), 7.50-7.31 (m, 2H), 7.25 (d, J = 15.7 Hz,1H), 4.43-4.19 (m, 2H), 3.71 (d, J = 42.3 Hz, 4H), 3.54-3.45 (m, 1H), 3.26 (d, J = 35.8 Hz, 3H). Example 3 Synthesis of 4-(4-fluoro-3-(4-(2-phenylquinoline-4-carbonyl)piperazine-1-carbonyl)benzyl)phthalazine-1(2H)-one (compound I-3) Compound 8 was synthesized according to the method of Example 1.

[0066] Compound 8 (50.0 mg, 0.136 mmol) was added to dichloromethane (3.00 mL), followed by 2-phenylquinoline-4-carboxylic acid (36.0 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol). After the reaction was complete, the mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 60:1 → 40:1 → 30:1). The fractions containing the target product were collected, combined, and concentrated under reduced pressure to obtain the target compound I-3 as a white or off-white solid (61.0 mg, yield: 79.9%). HRMS (m / z): [M+H] + C 36 H 29 FN5O3598.2249;found: 598.2241.

[0067] 1 H NMR (600 MHz, DMSO- d 6) δ 12.61 (d, J = 51.8 Hz, 1H), 8.33 (s, 3H), 8.22-8.14 (m, 2H), 7.93 (s, 1H), 7.87 (t, J = 7.3 Hz, 3H), 7.67 (d, J = 7.4 Hz,2H), 7.61-7.56 (m, 2H), 7.55-7.47 (m, 1H), 7.44-7.35 (m, 1H), 7.34-7.27 (m,1H), 7.16 (s, 1H), 4.38 (s, 1H), 4.27 (s, 1H), 3.86 (d, J = 85.6 Hz, 3H), 3.07(s, 1H), 2.79 – 2.68 (m, 4H). Example 4 Synthesis of 4-(4-fluoro-3-(4-(2-methylquinoline-6-carbonyl)piperazine-1-carbonyl)benzyl)phthalazine-1(2H)-one (compound I-4) Compound 8 was synthesized according to the method of Example 1.

[0068] Compound 8 (50.0 mg, 0.136 mmol) was weighed and dissolved in anhydrous dichloromethane (3.00 mL). 2-methylquinoline-6-carboxylic acid (27.5 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 h. After the reaction was complete, the mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 50:1 → 30:1). The fraction containing the target product was collected, combined, and concentrated under reduced pressure to obtain the target compound I-4, a white or off-white solid (54.0 mg, yield: 76.2%). HRMS (m / z): [M+H]⁺ calcd for C 31 H 27 FN5O3: 528.2093, found: 528.2086.

[0069] 1 H NMR (600 MHz, DMSO- d 6) δ 12.60 (s, 1H), 8.33-8.23 (m, 2H), 8.02 (s,1H), 7.96 (d, J = 8.7 Hz, 2H), 7.91-7.67 (m, 3H), 7.48 (d, J = 8.4 Hz, 1H), 7.42(d, J = 12.9 Hz, 1H), 7.41-7.33 (m, 1H), 7.28-7.16 (m, 1H), 4.32 (s, 2H), 3.81-3.55 (m, 4H), 3.49 (d, J = 14.0 Hz, 2H), 3.29 (s, 2H), 2.67 (s, 3H). Example 5 Synthesis of 4-(4-fluoro-3-(4-(8-hydroxyquinoline-7-carbonyl)piperazine-1-carbonyl)benzyl)phthalazine-1(2H)-one (compound I-5) Compound 8 was synthesized according to the method of Example 1.

[0070] Compound 8 (50.0 mg, 0.136 mmol) was dissolved in anhydrous dichloromethane (3.00 mL), followed by the sequential addition of 8-hydroxyquinoline-7-carboxylic acid (28.4 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol). The mixture was stirred at room temperature for 2 h. After the reaction was complete, the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 50:1 → 30:1). The fractions containing the target product were collected, combined, and concentrated under reduced pressure to obtain the target compound I-5 as a white or off-white solid (55.0 mg, yield: 77.4%). HRMS (m / z): [M+H]⁺calcd for C 30 H 25 FN5O4538.1890, found: 538.1885.

[0071] 1 H NMR (600 MHz, DMSO- d 6) δ 12.56 (d, J = 42.7 Hz, 1H), 8.91 (ddd, J =13.0, 4.2, 1.7 Hz, 1H), 8.41 (d, J = 1.9 Hz, 1H), 8.30-8.24 (m, 1H), 8.17-7.92(m, 2H), 7.89-7.80 (m, 1H), 7.73-7.52 (m, 4H), 7.46-7.36 (m, 1H), 7.35-7.23(m, 1H), 4.33 (s, 1H), 4.23 (s, 1H), 3.95-3.52 (m, 4H), 3.45 (s, 1H), 3.08(s, 2H), 2.87 (d, J = 5.3 Hz, 1H). Example 6 Synthesis of 4-(3-(4-(7-chloro-3-methylquinoline-8-carbonyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one (compound I-6) Compound 8 was synthesized according to the method of Example 1.

[0072] Compound 8 (50.0 mg, 0.136 mmol) was weighed and dissolved in anhydrous dichloromethane (3.00 mL). 7-chloro-3-methylquinoline-8-carboxylic acid (45.2 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 h. After the reaction was complete, the mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 50:1 → 40:1) as the eluent to give the target compound I-6 as a yellow or yellowish solid (61.0 mg, yield: 78.4%). HRMS (m / z): [M+H]⁺ calcd for C31H26ClFN5O3:570.1703, found: 570.1694.

[0073] 1 H NMR (600 MHz, DMSO-d6) δ 12.53 (d, J = 44.1 Hz, 1H), 8.77 (dd, J =8.3, 2.2 Hz, 1H), 8.21 (d, J = 43.5 Hz, 2H), 7.94 (s, 2H), 7.79 (s, 1H), 7.63(d, J = 8.8 Hz, 2H), 7.39 (s, 1H), 7.33-7.24 (m, 1H), 7.22 (s, 1H), 4.25 (d,J = 57.2 Hz, 2H), 3.90-3.59 (m, 3H), 3.47-3.37 (m, 1H), 3.30- 3.17 (m, 1H),3.16-2.97 (m, 2H), 2.83 (s, 1H), 2.43 (s, 3H). Example 7 Synthesis of 4-(3-(4-(3,7-dichloroquinoline-8-carbonyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one (compound I-7) Compound 8 was synthesized according to the method of Example 1.

[0074] Compound 8 (50.0 mg, 0.136 mmol) was dissolved in anhydrous DMF (3.00 mL), and 3,7-dichloroquinoline-8-carboxylic acid (49.3 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially. The mixture was stirred at room temperature for 2 h. After the reaction was complete, the mixture was extracted with ethyl acetate (20.0 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (volume ratio = 40:1 → 25:1) to give the target compound I-7 as a yellow or yellowish solid (62 mg, yield: 76.5%). HRMS (m / z): [M+H]⁺ calcd for C 30 H 23 Cl2FN5O3: 590.1240, found: 590.1232.

[0075] 1 H NMR (600 MHz, DMSO- d 6) δ 12.58 (d, J = 44.7 Hz, 1H), 8.96 (dd, J = 9.0, 2.5 Hz, 1H), 8.69 (dd, J = 11.1, 2.5 Hz, 1H), 8.30 (s, 2H), 8.07 (dd, J = 14.4,8.9 Hz, 1H), 8.01-7.89 (m, 1H), 7.87-7.78 (m, 2H), 7.48-7.41 (m, 1H), 7.39-7.31 (m, 1H), 7.21 (dt, J = 63.5, 8.9 Hz, 1H), 4.35 (s, 1H), 4.26 (s, 1H), 3.96-3.64 (m, 3H), 3.60-3.39 (m, 2H), 3.22-3.05 (m, 2H), 3.00-2.88 (m, 1H). Example 8 Synthesis of 4-(4-fluoro-3-(4-(6-fluoroquinoline-2-carbonyl)piperazine-1-carbonyl)benzyl)phthalazine-1(2H)-one (compound I-8) Compound 8 was synthesized according to the method of Example 1.

[0076] Compound 8 (50.0 mg, 0.136 mmol) was dissolved in anhydrous DMF (3.00 mL), followed by the addition of 6-fluoroquinoline-2-carboxylic acid (39.1 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol). The reaction was stirred at room temperature for 2 h. After the reaction was complete, the mixture was extracted with ethyl acetate (20.0 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (volume ratio = 40:1 → 30:1) to give the target compound I-8 as a yellow or yellowish solid (58.0 mg, yield: 78.7%). HRMS (m / z): [M+H]⁺ calcd for C 30 H 24 F2N5O3: 540.1847, found: 540.1841.

[0077] 1 H NMR (600 MHz, DMSO- d 6) δ 12.55 (d, J = 26.5 Hz, 1H), 8.45 (t, J = 9.5Hz, 1H), 8.29-8.12 (m, 2H), 8.06 (dd, J = 25.7, 4.0 Hz, 1H), 7.81 (ddt, J = 32.6,26.1, 8.1 Hz, 3H), 7.73-7.63 (m, 2H), 7.47-7.28 (m, 2H), 7.18 (dt, J = 38.1, 9.0 Hz, 1H), 4.28 (d, J = 31.6 Hz, 2H), 3.74 (s, 2H), 3.62 (d, J = 20.5 Hz, 2H), 3.53 (d, J = 5.2 Hz, 1H), 3.40 (s, 2H), 3.19 (s, 1H). Example 9 Synthesis of 4-(4-fluoro-3-(4-(2-(trifluoromethyl)quinoline-4-carbonyl)piperazine-1-carbonyl)benzyl)phthalazine-1(2H)-one (compound I-9) Compound 8 (50.0 mg, 0.136 mmol) was weighed and dissolved in anhydrous N,N-dimethylformamide (DMF, 3.00 mL). 2-trifluoromethylquinoline-4-carboxylic acid (49.3 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 h. After the reaction was completed as monitored by thin-layer chromatography (TLC), the aqueous phase was extracted with ethyl acetate (20.0 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 50:1 → 30:1). The fraction containing the target product was collected, combined, and concentrated under reduced pressure to give the target compound I-9 as a white or off-white solid (66.0 mg, yield: 82.3%). HRMS (m / z): [M+H]⁺ calcd for C 31 H 24 F4N5O3: 590.1815, found: 590.1812.

[0078] 1 H NMR (600 MHz, DMSO- d 6) δ 12.59 (d, J = 46.5 Hz, 1H), 8.33-8.23 (m,2H), 8.06 (d, J = 19.2 Hz, 1H), 8.02-7.96 (m, 2H), 7.86 (d, J = 8.2 Hz, 2H), 7.68(s, 1H), 7.50-7.35 (m, 2H), 7.27 (s, 1H), 7.15 (s, 1H), 4.31 (d, J = 57.7 Hz, 2H), 3.82 (d, J = 65.7 Hz, 3H), 3.66-3.41 (m, 2H), 3.27-3.16 (m, 1H), 3.05 (d, J = 23.2 Hz, 1H), 2.68 (s, 1H). Example 10 Synthesis of 4-(4-fluoro-3-(4-(quinoline-6-carbonyl)-1,4-diazacycloheptane-1-carbonyl)benzyl)phthalazine-1(2H)-one (compound I-10) Step (1): Synthesis of 4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)-1,4-diazacycloheptane-1-carboxylic acid tert-butyl ester (compound 5).

[0079] Compound 3 was synthesized according to the method in Example 1.

[0080] Compound 3 (2.00 g, 0.0067 mol) was dissolved in dichloromethane (40.00 mL). HATU (3.06 g, 0.0080 mol) and N,N-diisopropylethylamine (DIPEA, 2.60 g, 0.020 mol) were added sequentially under ice-water bath conditions. After stirring and activation for 30 min, N-Boc-1,4-diazacycloheptane (1.64 g, 0.0080 mol) was added. The ice-water bath was removed, and the mixture was allowed to warm naturally to room temperature while stirring for 2 h. After the reaction was complete, water was added to quench the reaction mixture. The mixture was extracted with dichloromethane, and the organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using petroleum ether / ethyl acetate (volume ratio = 1:1) to give compound 5 as a white or off-white solid (2.58 g, yield: 80.5%). HRMS (m / z): [M+H] + calcd for C 26 H 30 FN4O4481.2551;found: 481.2543.

[0081] Step (2): Synthesis of 4-(3-(1,4-diazacycloheptane-1-carbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one (compound 9) Compound 5 (2.00 g, 0.0043 mol) was dissolved in dichloromethane (10.00 mL), and trifluoroacetic acid (TFA, 2.00 mL) was slowly added dropwise at room temperature, with stirring for 1 h at room temperature. After the reaction was complete, the solvent was removed by vacuum distillation, and the residue was dissolved in dichloromethane and neutralized to weakly alkaline by slowly adding saturated sodium bicarbonate aqueous solution under ice-water bath conditions. Extraction was performed with dichloromethane (100 mL × 3), the organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by silica gel column chromatography with dichloromethane / methanol (v / v = 20:1) as the eluent to give compound 9 as a white or off-white solid (1.42 g, yield: 89.6%). HRMS (m / z): [M+H] +calcd for C 21 H 21 FN4O2381.1721; found: 381.1716.

[0082] Step (3): Synthesis of compound I-10 Compound 9 (50.0 mg, 0.136 mmol) was weighed and dissolved in anhydrous dichloromethane (3.00 mL). Quinoline-6-carboxylic acid (20.7 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 h. After the reaction was detected by thin-layer chromatography (TLC), the mixture was extracted with dichloromethane (20.0 mL × 3), and the organic phases were combined and washed sequentially with saturated sodium bicarbonate aqueous solution (20.0 mL × 1) and saturated sodium chloride aqueous solution (20.0 mL × 1). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 40:1 → 30:1) as the eluent to give the target compound I-10 as a white or off-white solid (54.30 mg, yield: 77.1%). HRMS (m / z): [M+H]⁺ calcd for C 31 H 27 FN5O3: 536.2092, found: 536.2087.

[0083] 1 H NMR (600 MHz, DMSO- d 6) δ 12.62 (d, J = 13.4 Hz, 1H), 9.07-8.77 (m,1H), 8.38 (d, J = 8.4 Hz, 1H), 8.27 (d, J = 44.7 Hz, 2H), 8.04 (d, J = 13.9 Hz, 1H), 7.96 (d, J = 7.0 Hz, 1H), 7.77 (d, J = 58.6 Hz, 3H), 7.59 (s, 1H), 7.42 (d, J= 25.3 Hz, 1H), 7.34-6.94 (m, 2H), 4.46-4.09 (m, 2H), 3.86 (s, 1H), 3.71 (d, J = 18.0 Hz, 3H), 3.57 (s, 1H), 3.47 (s, 1H), 3.32 (s, 2H), 1.62 (d, J = 42.3 Hz, 2H). Example 11 Synthesis of 4-(3-(4-(7-chloro-3-methylquinoline-8-carbonyl)-1,4-diazacycloheptane-1-carbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one (compound I-11) Compound 9 was synthesized according to the method of Example 10.

[0084] Compound 9 (50.0 mg, 0.136 mmol) was dissolved in dichloromethane (3.00 mL), and 7-chloro-3-methylquinoline-8-carboxylic acid (43.6 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially. The mixture was stirred at room temperature for 2 h. After the reaction was complete, the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography with dichloromethane / methanol (v / v = 40:1 → 30:1) as the eluent to give the target compound I-11 as a yellow solid (57.0 mg, yield: 74.3%). HRMS (m / z): [M+H]⁺ calcd for C 32 H 28 ClFN5O3: 585.1937, found: 585.1931.

[0085] 1 H NMR (600 MHz, Chloroform- d ) δ 11.65 (s, 1H), 8.77-8.56 (m, 1H), 8.46-8.21 (m, 1H), 7.85 (d, J = 5.7 Hz, 1H), 7.72 (dd, J = 13.6, 8.0 Hz, 2H),7.70-7.64 (m, 3H), 7.59 (s, 1H), 7.45 (d, J= 8.4 Hz, 1H), 7.01 (d, J = 1.8 Hz, 1H), 4.26 (d, J = 33.9 Hz, 2H), 4.15 (s, 2H), 3.97-3.72 (m, 2H), 3.53 (d, J = 7.3Hz, 1H), 3.44-3.29 (m, 2H), 3.17 (s, 1H), 2.44 (s, 1H), 2.41 (s, 4H). Example 12 Synthesis of N-(2-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoylamino)ethyl)quinoline-6-carboxamide (Compound I-12) Step (1): Synthesis of tert-butyl (2-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoylamino)ethyl)carbamate (compound 6).

[0086] Compound 3 was synthesized according to the method in Example 1.

[0087] Compound 3 (2.00 g, 0.0067 mol) was dissolved in dichloromethane (20.0 mL). HATU (3.06 g, 0.0080 mol) and N,N-diisopropylethylamine (DIPEA, 2.60 g, 0.020 mol) were added under ice-water bath conditions, and the mixture was stirred and activated for approximately 30 min. Subsequently, N-Boc-ethylenediamine (1.38 g, 0.0080 mol) was added, the ice-water bath was removed, and the mixture was allowed to warm naturally to room temperature. The reaction was continued with stirring at room temperature for 1 h. After the reaction was complete, the mixture was extracted with dichloromethane, and the organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give compound 6 as a white solid (2.64 g, yield: 89.1%). HRMS (m / z): [M+H] + calcd for C 23 H 26 FN4O4441.1933; found: 441.1927.

[0088] Step (2): Synthesis of N-(2-aminoethyl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide (compound 10) Compound 6 (2.00 g, 0.0043 mol) was dissolved in dichloromethane (10.0 mL), and trifluoroacetic acid (TFA, 2.00 mL) was slowly added dropwise at room temperature while stirring for 0.5 h. After the reaction was complete, the solvent was removed by vacuum distillation. The residue was dissolved in dichloromethane, and the solution was slowly added to a saturated sodium bicarbonate solution under ice-water bath conditions to adjust to a weakly alkaline state, followed by extraction with dichloromethane (100 mL × 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (v / v = 20:1) as the eluent to give compound 10 as a white or off-white solid (1.38 g, yield: 89.3%). HRMS (m / z): [M+H] + calcd for C 18 H 18 FN4O2341.1487; found: 341.1479.

[0089] Step (3): Synthesis of compound I-12 Compound 10 (50.0 mg, 0.136 mmol) was weighed and dissolved in anhydrous N,N-dimethylformamide (DMF, 3.00 mL). Quinoline-6-carboxylic acid (27.9 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 h. After the reaction was completed by thin-layer chromatography (TLC), the mixture was extracted with ethyl acetate (20.0 mL × 3). The organic phases were combined and washed sequentially with saturated sodium bicarbonate aqueous solution and saturated sodium chloride aqueous solution. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 50:1 → 40:1). The fraction containing the target product was collected, combined, and concentrated under reduced pressure to give the target compound I-12 as a white or off-white solid (55.0 mg, yield: 75.6%). HRMS (m / z): [M+H]⁺ calcd for C 28 H 23 FN5O3: 496.1779, found: 496.1772.

[0090] 1 H NMR (600 MHz, DMSO- d6) δ 12.60 (s, 1H), 8.97 (dd, J = 4.2, 1.7 Hz, 1H), 8.81 (t, J = 5.2 Hz, 1H), 8.50-8.39 (m, 3H), 8.31-8.24 (m, 1H), 8.16 (dd, J = 8.8, 2.0 Hz, 1H), 8.07 (d, J = 8.8 Hz, 1H), 7.94 (d, J = 8.0 Hz, 1H), 7.87-7.83(m, 1H), 7.81 (d, J = 7.6 Hz, 1H), 7.63 (dd, J = 6.9, 2.2 Hz, 2H), 7.47-7.41 (m,1H), 7.20 (dd, J = 10.3, 8.5 Hz, 1H), 4.31 (s, 2H), 3.48 (d, J = 4.0 Hz, 4H). Example 13 Synthesis of 7-chloro-N-(2-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoylamino)ethyl)-3-methylquinoline-8-carboxamide (Compound I-13) Compound 10 was synthesized according to the method of Example 12.

[0091] Compound 10 (50.0 mg, 0.136 mmol) was weighed and dissolved in anhydrous N,N-dimethylformamide (DMF, 3.00 mL). 7-chloro-3-methylquinoline-8-carboxylic acid (21.6 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 h. After the reaction was completed as monitored by thin-layer chromatography (TLC), the mixture was extracted with ethyl acetate (20.0 mL × 3). The combined organic phases were washed sequentially with saturated sodium bicarbonate aqueous solution and saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 50:1 → 40:1). The fraction containing the target product was collected, combined, and concentrated under reduced pressure to give the target compound I-13 as a yellow solid (61.0 mg, yield: 76.3%). HRMS (m / z): [M+H]⁺ calcd for C 29 H 24 ClFN5O3: 554.1546, found: 554.1541.

[0092] 1 H NMR (600 MHz, DMSO- d 6) δ 12.54 (s, 1H), 8.56-8.47 (m, 2H), 8.26-8.17 (m, 2H), 8.13-8.07 (m, 1H), 7.90-7.86 (m, 2H), 7.74 (d, J = 1.2 Hz, 2H),7.64-7.55 (m, 2H), 7.42 (ddd, J = 7.9, 4.5, 2.3 Hz, 1H), 7.17 (dd, J = 10.4, 8.5Hz, 1H), 4.25 (s, 2H), 3.44 (s, 4H), 2.35 (s, 3H). Example 14 Synthesis of N-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoylamino)butyl)quinoline-6-carboxamide (Compound I-14) Step (1): Synthesis of tert-butyl(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoylamino)butyl)carbamate (compound 7) Compound 3 was synthesized according to the method in Example 1.

[0093] Compound 3 (2.00 g, 0.0067 mol) was dissolved in anhydrous dichloromethane (20.0 mL). HATU (3.06 g, 0.0080 mol) and N,N-diisopropylethylamine (DIPEA, 2.60 g, 0.020 mol) were added under ice-water bath conditions, and the mixture was stirred and activated for approximately 30 min. Subsequently, N-Boc-butanediamine (1.52 g, 0.0080 mol) was added, the ice-water bath was removed, and the mixture was allowed to warm naturally to room temperature. The reaction was continued with stirring at room temperature for 1 h. After the reaction was complete, the mixture was extracted with dichloromethane, and the organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give the target compound 7 as a white solid (2.78 g, yield: 88.5%). HRMS (m / z): [M+H] + calcd for C 25 H 30 FN4O4469.2246; found: 469.2240.

[0094] Step (2): Synthesis of N-(4-aminobutyl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide (compound 11) Compound 7 (2.00 g, 0.0043 mol) was dissolved in dichloromethane (10.0 mL), and trifluoroacetic acid (TFA, 2.00 mL) was slowly added at room temperature, with the mixture stirred for 0.5 h at room temperature. After the reaction was complete, the solvent was removed by vacuum distillation. The residue was dissolved in dichloromethane, and the solution was adjusted to weakly alkaline by slow addition of saturated sodium bicarbonate solution under ice-water bath conditions, followed by extraction with dichloromethane (100 mL × 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography with dichloromethane / methanol (v / v = 20:1) as the eluent to give compound 11, which was a white or off-white solid (1.42 g, yield: 90.2%). HRMS (m / z): [M+H] + calcd for C 20 H 22 FN4O2369.1721; found: 369.1716.

[0095] Step (3): Synthesis of compound I-14 Compound 11 (50.0 mg, 0.136 mmol) was weighed and dissolved in anhydrous N,N-dimethylformamide (DMF, 3.00 mL). Quinoline-6-carboxylic acid (25.8 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 h. After the reaction was completed by thin-layer chromatography (TLC), the mixture was extracted with ethyl acetate (20.0 mL × 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 50:1 → 40:1). The fraction containing the target product was collected, combined, and concentrated under reduced pressure to give the target compound I-14 as a white or off-white solid (56.0 mg, yield: 78.9%). HRMS (m / z): [M+H]⁺calcd for C 30 H 27 FN5O3: 524.2092, found: 524.2086.

[0096] 1 H NMR (600 MHz, DMSO-d6) δ 12.54 (s, 1H), 8.95 (ddd, J = 36.8, 4.2,1.8 Hz, 1H), 8.67 (d, J = 1.9 Hz, 1H), 8.43 (d, J = 2.0 Hz, 2H), 8.26-8.17(m, 2H), 8.10 (d, J = 2.0 Hz, 1H), 8.02 (s, 1H), 7.90 (s, 1H), 7.82 (s, 1H),7.76 (d, J = 1.2 Hz, 1H), 7.61-7.49 (m, 2H), 7.37 (td, J = 4.7, 4.2, 2.5 Hz,1H), 7.14 (d, J = 1.8 Hz, 1H), 4.26 (s, 2H), 3.88 (s, 1H), 3.21 (d, J = 6.1Hz, 3H), 1.59 – 1.46 (m, 4H). Example 15 Synthesis of 7-chloro-N-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoylamino)butyl)-3-methylquinoline-8-carboxamide (Compound I-15) Compound 11 was synthesized according to the method of Example 14.

[0097] Compound 11 (50.0 mg, 0.136 mmol) was weighed and dissolved in anhydrous N,N-dimethylformamide (DMF, 3.00 mL). 7-chloro-3-methylquinoline-8-carboxylic acid (33.03 mg, 0.150 mmol), HATU (62.0 mg, 0.163 mmol), and N,N-diisopropylethylamine (DIPEA, 70.0 mg, 0.542 mmol) were added sequentially, and the mixture was stirred at room temperature for 2 h. After the reaction was completed as monitored by thin-layer chromatography (TLC), the mixture was extracted with ethyl acetate (20.0 mL × 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (volume ratio = 50:1 → 40:1). The fraction containing the target product was collected, combined, and concentrated under reduced pressure to give the target compound I-15 as a white or off-white solid (58.0 mg, yield: 74.7%). HRMS (m / z): [M+H]⁺ calcd for C 31 H 28 ClFN5O3: 572.1860, found: 572.1856.

[0098] 1 H NMR (600 MHz, DMSO- d 6) δ 12.56 (s, 1H), 8.76 (d, J = 2.2 Hz, 1H), 8.38 (t, J = 5.7 Hz, 1H), 8.31-8.17 (m, 3H), 8.12 (dd, J = 2.3, 1.2 Hz, 1H),7.94-7.86 (m, 2H), 7.77 (d, J = 1.1 Hz, 1H), 7.59-7.52 (m, 2H), 7.42-7.38 (m,1H), 7.16 (dd, J = 10.2, 8.5 Hz, 1H), 4.28 (s, 2H), 3.29 (s, 2H), 3.24 (d, J =6.4 Hz, 2H), 2.46 (s, 3H), 1.66-1.59 (m, 2H), 1.59-1.52 (m, 2H). Given that the number of compounds in this invention is large, and that the compounds have the same core pharmacodynamic skeleton, differing only in the linker arm or quinoline carboxylic acid substituents, and that the synthesis methods are highly consistent, representative compounds are selected for pharmacological evaluation. The results can reasonably reflect the overall bioactivity trend of the same series of compounds.

[0099] The technical solution of the present invention will be further illustrated by the following experiments. The compounds tested in the following experimental examples were all prepared using the methods described in the above embodiments.

[0100] Experiment Example 1: Inhibitory Effects of Representative Compounds Linked by Different Linkers on the Proliferation of BRCA1 Gene-Mutant and Wild-Type Breast Cancer Cells I. Experimental Methods The MTT assay was used to evaluate the inhibitory effects of representative compounds linked by different linkers in this invention on the proliferation of different breast cancer cell lines and normal breast cells MCF-10A. HCC1937 (BRCA1-null, human triple-negative breast cancer cells), HCC1806 (human triple-negative breast cancer cells), MCF-7 (human breast cancer cells), MDA-MB-436 (BRCA1-null, human triple-negative breast cancer cells), and 4T1 cells (mouse triple-negative breast cancer cells) were digested and counted in complete culture medium (containing 10% FBS) during the logarithmic growth phase. The cells were seeded in 96-well plates at 4000 cells / well for HCC1806, MCF-7, MDA-MB-436, and 4T1, and 3000 cells / well for HCC1937 and MCF-10A. After culturing for 24 h, the supernatant was removed, and the drugs were prepared as a 10 μmol / mL DMSO stock solution, which was then diluted with complete culture medium to different concentrations. Subsequently, the supernatant was removed, and complete culture medium containing different concentrations of the drug was added for further incubation for 72 h. A blank control group and an olaparib positive control group were also set up, with three replicates for each concentration. After co-culture, the supernatant was removed, and 100 μL of MTT solution (5 mg / mL) was added. After incubation at 37°C for 2 h, the supernatant was removed again, and 100 μL of DMSO solution was added and thoroughly shaken to dissolve the crystals. The absorbance was measured at 490 nm using a microplate reader, and the half-maximal inhibitory concentration (IC50) of the drug against tumor cells was fitted using GraphPad Prism.

[0101] The structural formula of olaparib is as follows: .

[0102] II. Experimental Results The experimental results are shown in Table 1. The results indicate that, among the tested linker arms, compounds with piperazine linker arms generally exhibited superior cell proliferation inhibitory activity. The piperazine ring showed the strongest inhibitory effect against all five breast cancer cell types, making it the optimal linker arm group.

[0103] Among other compounds with linker groups, compounds I-11, I-12, I-13, and I-15 also showed enhanced proliferative inhibitory activity against various breast cancer cell types compared to olaparib, especially I-11 and I-12, which showed significantly enhanced inhibitory effects against all five types of breast cancer cell types.

[0104] Table 1. IC50 of representative compounds with different linker arm structures on different breast cancer cell lines. Note: The values ​​in the table are IC50 values, in nmol / mL, i.e. μM; L and R refer to the L group and R group in Formula I, respectively.

[0105] The above results indicate that the piperazine ring is the optimal linker group; compared with olaparib, compounds I-1, I-2, I-6, I-11, and I-12 all showed significantly enhanced proliferative inhibition against the five types of breast cancer cells.

[0106] Experimental Example 2: Inhibitory Effects of Representative Compounds with Different Derivative Groups on the Proliferation of BRCA1 Mutant and Wild-Type Cells I. Experimental Methods The MTT assay was used to evaluate the inhibitory effects of representative compounds with different derivative groups in this invention on the proliferation of different breast cancer tumor cell lines and normal breast cells MCF-10A. The drugs tested in this experiment were compounds I-3, I-4, I-5, I-7, I-8, I-9, and olaparib.

[0107] HCC1937 (BRCA1-null), HCC1806, MCF-7, MDA-MB-436 (BRCA1-null), and 4T1 cells were digested and counted in complete medium (containing 10% FBS) during the logarithmic growth phase. Cells were seeded in 96-well plates at 4000 cells / well for HCC1806, MCF-7, MDA-MB-436, and 4T1, and 3000 cells / well for HCC1937 and MCF-10A. After 24 h of culture, the supernatant was removed, and the drug was prepared as a 10 μmol / mL DMSO stock solution, diluted with complete medium to different concentrations. Subsequently, the supernatant was removed, and complete medium containing different drug concentrations was added, and the cells were cultured for another 72 h. A blank control group and an olaparib positive control group were also included, with triplet wells for each concentration. After co-culture, the supernatant was removed, and 100 μL of MTT solution (5 mg / mL) was added. After incubation at 37°C for 2 h, the supernatant was removed, and 100 μL of DMSO solution was added and shaken thoroughly to dissolve the crystals. The absorbance value was measured at 490 nm using an ELISA reader, and the half-maximal inhibitory concentration (IC50) of the drug against tumor cells was fitted using GraphPad Prism.

[0108] II. Experimental Results The experimental results are shown in Table 2. The results show that, compared with olaparib, the compounds in this embodiment exhibited significantly enhanced cell proliferation inhibitory activity against various breast cancer cell models. In particular, compounds I-3, I-7, I-8, and I-9 showed enhanced inhibitory activity against all breast cancer cell types, with compounds I-3 and I-9 exhibiting the best cell proliferation inhibitory activity.

[0109] Table 2. Effects of R-group derivatization on IC50 of different breast cancer cell lines. Note: The values ​​in the table are IC50 values, in nmol / mL, i.e., μM.

[0110] Experiment Example 3: Cell Proliferation Assay I. Experimental Methods MCF-7 and 4T1 cells were seeded into 12-well plates, each well with a coverslip, and cultured for 48 hours with different concentrations of compound I-9. EdU staining was performed using BeyoClick™ EdU cell proliferation kit (containing Alexa Fluor 47, Shanghai, China). Cells were co-incubated with 10 μM EdU for 2 hours, then fixed with 4% paraformaldehyde solution for 15 minutes and stained with Click staining solution. Finally, cells were stained with DAPI and observed under a confocal microscope.

[0111] II. Experimental Results The results are as follows Figure 1 As shown in the figure. Compared with the control group, after 48 hours of compound treatment, the proportion of EdU-positive cells in MCF-7 cells and 4T1 cells was significantly reduced, and the proportion decreased with increasing drug concentration. This indicates that the number of cells in the proliferative phase gradually decreased with increasing drug concentration, and that compound I-9 can inhibit cell proliferation in a concentration-dependent manner.

[0112] As can be seen from the above embodiments and experimental examples, this invention synthesizes a novel PARP inhibitor derivative. This derivative uses the clinical PARP inhibitor olaparib as the core pharmacophore and is constructed by covalent coupling with a quinoline carboxylic acid derivative. The derivative is the compound shown in formula (I) and its pharmaceutically acceptable salt, solvate, or hydrate, and its structure includes: a first structural unit with olaparib as the core skeleton, a linker arm L, and a quinoline carboxylic acid fragment R; wherein the structural types and substituents of the linker arm L and the quinoline carboxylic acid fragment R are adjustable. This invention also provides a method for preparing the compound, which is simple in steps, highly versatile, and facilitates structural expansion, making it suitable for constructing compound libraries with different combinations of linkers and quinoline carboxylic acid fragments. The compound can be used to prepare drugs or drug compositions for inhibiting tumor cell proliferation and treating tumor-related diseases, showing excellent application prospects.

Claims

1. A PARP inhibitor-quinoline carboxylic acid conjugate, or its optical isomer, or its tautomer, or its deuterated compound, or its tritated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form, characterized in that, The structure of the PARP inhibitor-quinoline carboxylic acid coupling derivative compound is shown in Formula I: in, L is selected from -NH- L x -NH-、 ; L x Selected from C 2-4 Alkylene; m is 1 or 2; R is -C(O)R1; R1 is selected from the substituted or unsubstituted quinoline ring of R2, and R2 is selected from halogen, -OH, C 1-6 Alkyl, C 1-6 Haloalkyl, C 6-15 At least one of the aromatic rings.

2. The PARP inhibitor-quinoline carboxylic acid conjugate, or its optical isomer, or its tautomer, or its deuterated compound, or its tritated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form according to claim 1, is characterized in that: L is selected from -NH- L x -NH-、 ; L x Selected from C 2-4 Alkylene; m is 1 or 2; R is -C(O)R1; R1 is selected from a quinoline ring that is substituted or unsubstituted by R2, and R2 is selected from at least one of F, Cl, -OH, methyl, benzene ring, and trifluoromethyl.

3. The PARP inhibitor-quinoline carboxylic acid conjugate, or its optical isomer, or its tautomer, or its deuterated compound, or its tritated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form according to claim 2, is characterized in that: The number of R2 is 0, 1, or 2; The linker site of R1 to the carbonyl group is located at position 2, 4, 6, 7 or 8 of the quinoline ring; The connection site between R1 and R2 is located at at least one of the positions 2, 3, 6, 7, and 8 of the quinoline ring.

4. The PARP inhibitor-quinoline carboxylic acid conjugate, or its optical isomer, or its tautomer, or its deuterated compound, or its tritated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form according to claim 3, is characterized in that: L is selected from , , , One of them; L is In this case, R1 is selected from the quinoline ring with or without R2 substitution, and the linking site of R1 to the carbonyl group is located at position 2, 4, 6, 7 or 8 of the quinoline ring. R2 is selected from phenyl, methyl, Cl, F, trifluoromethyl, and the number of R2 is 0, 1 or 2. The linking site of R1 and R2 is located at at least one of the positions 2, 3, 6, 7 or 8 of the quinoline ring. L is or In this case, R1 is selected from the quinoline ring substituted with R2, the linking site of R1 to the carbonyl group is located at the 6 or 8 position of the quinoline ring, R2 is at least one of methyl and Cl, the number of R2 is 0 or 2, and the linking site of R1 to R2 is located at at least one of the 3 or 7 positions of the quinoline ring. L is In this case, R1 is selected from the quinoline ring that is substituted or unsubstituted by R2, the linking site of R1 to the carbonyl group is located at the 6th or 8th position of the quinoline ring, R2 is at least one of methyl and Cl, the number of R2 is 0 or 2, and the linking site of R1 to R2 is located at at least one of the 3rd or 7th position of the quinoline ring.

5. The PARP inhibitor-quinoline carboxylic acid conjugate, or its optical isomer, or its tautomer, or its deuterated compound, or its tritated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form according to claim 4, is characterized in that: L is R1 is selected from the quinoline ring substituted with R2, and the linking site of R1 to the carbonyl group is located at the 4 position of the quinoline ring. R2 is selected from phenyl or trifluoromethyl, and the linking site of R1 to R2 is located at the 2 position of the quinoline ring.

6. The PARP inhibitor-quinoline carboxylic acid conjugate, or its optical isomer, or its tautomer, or its deuterated compound, or its tritated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form according to claim 3, is characterized in that: R is selected from one of the following structures: 。 7. The PARP inhibitor-quinoline carboxylic acid conjugate, or its optical isomer, its tautomer, its deuterated compound, its tritated compound, its pharmaceutically acceptable salt, its prodrug, its hydrate, its solvate, or its crystal form according to any one of claims 1-6, characterized in that, The structural formula of the PARP inhibitor-quinoline carboxylic acid coupling derivative compound is shown in any of the following: 。 8. A method for preparing the PARP inhibitor-quinoline carboxylic acid coupling derivative compound, or its optical isomer, or its tautomer, or its deuterated compound, or its tritated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form as described in any one of claims 1-7, characterized in that, It includes: ； Compound a reacts with ROH via a condensation reaction to produce the compound shown in Formula I.

9. The method for preparing the PARP inhibitor-quinoline carboxylic acid coupling derivative, or its optical isomer, or its tautomer, or its deuterated compound, or its tritated compound, or its pharmaceutically acceptable salt, or its prodrug, or its hydrate, or its solvate, or its crystal form according to claim 8, characterized in that: The condensation reaction is carried out under the action of a condensing agent and a base. The condensing agent is selected from HATU, HBTU, DCC, EDCI or CDI, the base is selected from diisopropylethylamine, triethylamine or DMAP, and the solvent for the condensation reaction is selected from dichloromethane, N,N-dimethylformamide, tetrahydrofuran or acetonitrile. And / or, compound a is prepared by the following method: Step 1: 2-Formylbenzoic acid is mixed with dimethyl phosphite and reacted under the action of an alkoxide base to obtain compound 1; Step 2: Compound 1 is mixed with an organic base and 2-fluoro-5-formylbenzonitrile, and the reaction yields compound 2; Step 3: Compound 2 undergoes an alkaline conversion reaction, and then hydrazine hydrate is added to continue the reaction to obtain compound 3; Step 4: Compound 3 undergoes a condensation reaction with R3LH to obtain compound b, where R3 is an amino protecting group; Step 5: Compound b undergoes a deprotection reaction to obtain compound a.

10. Use of the PARP inhibitor-quinoline carboxylic acid coupling derivative compound of any one of claims 1-7, or its optical isomer, tautomer, deuterated compound, tritated compound, pharmaceutically acceptable salt, prodrug, hydrate, solvate, or crystal form in the preparation of an anti-breast cancer drug.

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