Quinoxalinone derivative targeting poly ADP ribose polymerase and androgen receptor as well as preparation method and application of quinoxalinone derivative

Abstract

The invention relates to a quinoxalinone derivative targeting poly ADP ribose polymerase and an androgen receptor as well as a preparation method and application of the quinoxalinone derivative. The structural formula of the quinoxalinone derivative is as follows: # imgabs0 #, R1 is one of hydrogen, methoxyl, methyl and halogen, the substituent position of R1 is at least one of 5, 6, 7 and 8 sites of a quinoxalinone ring, and X is a carbon atom or a nitrogen atom. The quinoxalinone derivative provided by the invention can be used as an active photosensitizer, a PARP inhibitor and an androgen receptor antagonist and can induce cell synthesis and death, and the process can improve the oxidation state and oxygen concentration in cells and improve the hypoxic microenvironment in tumors, so that the photodynamic therapy effect is further enhanced, namely, the effect of the quinoxalinone derivative is improved. The PDT therapy and the PARP inhibitor have a synergistic effect.

Description

Technical Field

[0001] The present invention relates to the technical field of novel drug development, and in particular to a quinoxalinone derivative targeting poly (ADP ribose) polymerase and androgen receptor, and a preparation method and application thereof. Background Art

[0002] Photodynamic therapy (PDT) is a treatment method that uses light of a specific wavelength to excite photosensitive molecules, inducing a "photodynamic" effect, thereby achieving anti-tumor effects. During the PDT process, photosensitive drugs generate highly active reactive oxygen species (ROS) through energy or electron transfer under light. These ROS can react with biological macromolecules in cells, causing oxidative damage, thereby inducing cytotoxicity and killing tumor cells.

[0003] Compared with traditional treatments, PDT has significant advantages, mainly in its ability to use the high temporal and spatial resolution of light for precise treatment, thereby reducing side effects. However, high concentrations of reduced glutathione in tumor cells can quickly scavenge ROS free radicals produced by photosensitive molecules, significantly weakening their efficacy. In addition, the generation of ROS by photosensitive molecules depends on the presence of molecular oxygen (O2), and the microenvironment of many solid tumors is hypoxic, which greatly reduces the therapeutic effect of PDT in hypoxic areas within the tumor.

[0004] Poly (ADP-ribose) polymerase (PARP) is a type of ribozyme with glycosidase activity. It mainly modifies target molecules through poly (ADP-ribosylation) and participates in important biological processes such as DNA repair, transcriptional regulation, chromatin dynamics, hypoxia response, metabolic regulation, cell death and genome stability. The PARP family includes 18 members, among which PARP1 is the most critical member, responsible for about 80-90% of PARylation reactions in cells. In the process of DNA damage repair, PARP1 mainly acts as a sensor. When DNA breaks, the zinc finger structure of PARP1 can quickly bind to the damaged site and recruit repair-related proteins through poly (ADP-ribosylation) to initiate DNA repair. The catalytic activity of PARP1 is activated after DNA damage, promoting PARylation modification reactions, synthesizing PAR chains, modifying itself and target molecules such as histones and non-histones, thereby regulating the DNA repair process. Therefore, PARP1 plays a vital role in the DNA repair process, and the absence of the PARP1 gene will lead to the failure of the base excision repair (BER) pathway. If PARP1 is defective in function, DNA damage will accumulate, eventually causing cell cycle arrest and possibly cell death, which makes PARP an important target in tumor treatment and promotes the research and development of PARP inhibitors.

[0005] PARP inhibitors inhibit the detachment of PARP protein from the DNA damage site by binding to the catalytic site of PARP1 or PARP2. PARP bound to DNA will cause replication fork stalling during DNA replication, hindering the normal progress of DNA replication, thereby inducing cell cycle arrest and ultimately leading to cell death. Currently, PARP inhibitors approved for clinical use include Olaparib, Niraparib, Fluzoparib and Pamiparib, which are mainly used for the treatment or maintenance treatment of tumors such as ovarian cancer, breast cancer and prostate cancer.

[0006] Prostate cancer is the most common malignant tumor in the male reproductive system. Androgens and their receptors (AR) play a vital role in the occurrence and development of prostate cancer. Therefore, androgen receptors and their related pathways have become key targets for the treatment of prostate cancer. Androgen receptor antagonists or anti-androgen drugs compete with testosterone (T) or dihydrotestosterone (DHT) to bind to the ligand binding pocket of AR, block the activation of AR signaling pathways, inhibit the transcription of downstream target genes, and prevent the synthesis of proteins required for tumor cell growth, thereby inhibiting the progression of prostate cancer. According to their chemical structures, AR antagonists can be divided into two categories: steroidal and non-steroidal. Steroidal AR antagonists are usually synthetic hydroxyprogesterone derivatives. Representative drugs include cyproterone acetate (CPA), which has a steroidal skeleton similar to DHT. Although these drugs have a high affinity for binding to AR, their clinical application is limited to a certain extent due to the possibility of cardiovascular and liver side effects. Non-steroidal AR antagonists are more widely used in clinical practice and have more significant therapeutic effects. The approved nonsteroidal AR antagonists on the market mainly target the C-terminal ligand binding region (LBP) of AR and exert their anti-tumor effects by blocking the signal transduction of AR. The first generation of nonsteroidal AR antagonists include flutamide, nilutamide and bicalutamide, while the second generation of nonsteroidal AR antagonists include enzalutamide, apalutamide and darolutamide.

[0007] (Darolutamide). Although the first generation of non-steroidal AR antagonists have shortcomings such as weak affinity and easy to induce drug resistance, the second generation of drugs have overcome these shortcomings and shown better clinical effects. Apalutamide is an analogue of enzalutamide. Both contain the core structure of benzothiohydantoin. Studies have shown that these drugs can not only competitively bind to AR, but also inhibit the nuclear localization function of AR and interfere with the binding of AR complex to DNA, thereby enhancing its anti-tumor effect.

[0008] The progress of photodynamic therapy (PDT), the research and development of PARP inhibitors and androgen receptor antagonists have greatly promoted the progress of malignant tumor treatment. However, there are still many challenges in clinical application. Specifically, during the treatment of photosensitizers, the high concentration of reduced glutathione in tumor cells will quickly remove the reactive oxygen free radicals produced by photosensitive molecules under light, thereby weakening the therapeutic effect. In addition, the hypoxic microenvironment inside the tumor also has a significant effect on the efficacy of photodynamic therapy. Compared with traditional chemotherapy drugs, targeted small molecule inhibitors usually cause fewer side effects and are better tolerated by patients by inhibiting the function of key proteins in tumor cells. However, off-target effects and drug resistance are still difficult problems that need to be solved in the field of tumor treatment. Summary of the invention

[0009] In order to solve the technical problems of poor effect of traditional photosensitizers and limitations of small molecule targeted drugs in the prior art, a bioactive photosensitizer that can induce synthetic lethality of tumor cells and efficiently generate ROS is designed and synthesized by finding key protein targets related to DNA repair in tumor cells, thereby improving the effect of photodynamic therapy. Therefore, the present invention provides a quinoxalinone derivative targeting poly ADP ribose polymerase and androgen receptor, and a preparation method and application thereof.

[0010] The first object of the present invention is to provide a quinoxalinone derivative targeting poly (ADP ribose) polymerase and androgen receptor, wherein the structural formula of the quinoxalinone derivative is as follows:

[0011]

[0012] Among them, R 1 is hydrogen, methoxy, methyl or halogen, the substituent position of R1 is at least one of the 5, 6, 7, 8 positions of the quinoxalinone ring, and X is a carbon atom or a nitrogen atom.

[0013] In some embodiments of the present invention, the halogen is selected from F, Cl, Br, and I.

[0014] The second object of the present invention is to provide a method for preparing the quinoxalinone derivative, comprising the following steps:

[0015] (1) Compound 1b, 6-azidohexanoic acid, a condensation agent, a catalyst, and an organic base are used to carry out an amidation condensation reaction to prepare compound 1c;

[0016] (2) Compound 1e, 2,5-thiophenedicarboxaldehyde and an organic base are subjected to a condensation reaction to prepare compound 1f;

[0017] (3) Compound 1c, compound 1f, a copper catalyst, and a reducing agent were used to prepare compound 1g;

[0018] (4) Compound 1g, compound 1j, an organic base and a catalyst are used to prepare compound 1, i.e., a quinoxalinone derivative; wherein the structures of compound 1b, compound 1e, compound 1f, compound 1c, compound 1g, compound 1j and compound 1 are as follows:

[0019]

[0020]

[0021] In some embodiments of the present invention, in step (1), the molar ratio of the compound 1b, 6-azidohexanoic acid, condensing agent, catalyst, and organic base is 1: (1-2): (1-2): (1-0.5): (1-5);

[0022] Reaction conditions: reaction temperature is room temperature, reaction time is 10 to 12 hours.

[0023] In some embodiments of the present invention, in step (1), the catalyst is selected from 1-hydroxybenzotriazole and / or 4-dimethylaminopyridine;

[0024] The condensing agent is selected from one or more of O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, dicyclohexylcarbodiimide, diisopropylcarbodiimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide;

[0025] The organic base is selected from one or more of N,N-diisopropylethylamine, triethylamine and ethylenediamine.

[0026] In some embodiments of the present invention, in step (1), compound 1b is prepared by the following method: compound 1a, 2,3-diaminobenzamide, a condensation agent and an organic base are subjected to an amidation condensation reaction and then the amino group protection is removed to obtain compound 1b.

[0027] Furthermore, the molar ratio of the compound 1a, 2,3-diaminobenzamide, the condensation agent and the organic base is 1:(1-2):(1-2):(1-3).

[0028] Further, the condensing agent is selected from one or more of O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, dicyclohexylcarbodiimide, diisopropylcarbodiimide, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide;

[0029] The organic base is selected from one or more of N,N-diisopropylethylamine, triethylamine and ethylenediamine.

[0030] In some embodiments of the present invention, in step (2), the molar ratio of compound 1e, 2,5-thiophenedicarboxaldehyde, and organic base is 1:(1-3):(1-5);

[0031] Organic bases include pyridine and / or imidazole;

[0032] Reaction conditions: reaction temperature 100-120°C, reaction time 6-12 hours.

[0033] In some embodiments of the present invention, in step (2), compound 1d It reacts with 3-bromopropyne and an inorganic base under alkaline conditions to undergo nucleophilic substitution to form compound 1e.

[0034] Furthermore, compound 1d The molar ratio of 3-bromopropyne to inorganic base is 1:(1-2):(1-3).

[0035] In some embodiments of the present invention, in step (3), the molar ratio of compound 1c, compound 1f, copper catalyst, and reducing agent is 1:(1-2):(1-2):(1-2); the copper catalyst is selected from one or more of copper sulfate, copper (II) bromide, copper (II) trifluoromethanesulfonate, and copper (II) acetate;

[0036] The reducing agent is selected from sodium ascorbate;

[0037] Reaction conditions: reaction temperature is room temperature, reaction time is 20 to 24 hours.

[0038] In some embodiments of the present invention, in step (4), the molar ratio of compound 1g, compound 1j, organic base, and catalyst is 1:(1-3):(1-3):(1-0.5);

[0039] The organic base includes one or more of tetrahydropyrrole, hexahydropyridine and piperazine;

[0040] Reaction conditions: reaction temperature 50-60°C, reaction time 2-4 hours.

[0041] In some embodiments of the present invention, in step (4), the compound 1j is prepared by the following method: Compound 1h reacts with compound 1i and an organic base under alkaline conditions to undergo a cyclization reaction to form compound (1j):

[0042]

[0043] Furthermore, the molar ratio of compound 1h, compound 1i and the organic base is 1:(1-2):(1-3).

[0044] The third object of the present invention is to provide the use of the quinoxalinone derivative in the preparation of a photosensitizer for photodynamic therapy.

[0045] The above technical solution of the present invention has the following advantages compared with the prior art:

[0046] 1) The active photosensitizer targeting PARP and androgen receptor provided by the present invention can inhibit the activity of poly (ADP-ribose) polymerase, hinder the DNA repair of tumor cells, induce tumor cell death, and thus improve the photodynamic therapy effect.

[0047] 2) The active photosensitizer targeting PARP and androgen receptor provided by the present invention can competitively bind to androgen receptor, inhibit the nuclear localization function of androgen receptor, and interfere with the binding of androgen receptor complex to DNA, thereby improving the photodynamic therapy effect.

[0048] 3) The active photosensitizer targeting PARP and androgen receptor provided by the present invention has a strong ability to generate reactive oxygen free radicals and can generate reactive oxygen species in the hypoxic tumor microenvironment.

[0049] 4) The active photosensitizer targeting PARP and androgen receptor provided by the present invention has a high killing effect on prostate cancer cells and has very low dark toxicity to normal cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] In order to make the content of the present invention more clearly understood, the present invention is further described in detail below according to specific embodiments of the present invention in conjunction with the accompanying drawings, wherein:

[0051] Figure 1 This is the mass spectrometry characterization of compound 1 in the example of the present invention.

[0052] Figure 2 This is the nuclear magnetic resonance hydrogen spectrum characterization of compound 1 in the example of the present invention.

[0053] Figure 3 It is the ultraviolet absorption spectrum of compound 1 in the embodiment of the present invention.

[0054] Figure 4 It is the fluorescence spectrum of compound 1 in the embodiment of the present invention.

[0055] Figure 5 This is a characterization diagram of ROS generated by compound 1 in the example of the present invention under blue light excitation.

[0056] Figure 6 This is the cellular uptake of compound 1 in the examples of the present invention.

[0057] Figure 7This is a diagram of intracellular ROS detection of cells to compound 1 in an example of the present invention.

[0058] Figure 8 This is a study on the toxicity of compound 1 on prostate cells in the examples of the present invention. DETAILED DESCRIPTION

[0059] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

[0060] The present invention also provides a method for preparing a quinoxalinone derivative targeting poly (ADP ribose) polymerase and androgen receptor, comprising the following steps:

[0061]

[0062] The specific steps include:

[0063] Step 1: Compound (1c) and compound (1f) undergo click cyclization reaction to obtain compound (1g).

[0064] In step 1, copper sulfate and sodium ascorbate are also added; the molar ratio of compound (1f), compound (1c), copper sulfate, and sodium ascorbate is 1:(1-2):(1-2):(1-2). As an example, an azide derivative (compound 1c) and an alkyne derivative (compound 1f) are dispersed in a mixed solvent of tetrahydrofuran and water (the volume ratio of tetrahydrofuran to water is 8:1), copper sulfate and sodium ascorbate are added, and stirred at room temperature for 20 to 24 hours. After the reaction is completed, the organic solvent in the reaction solution is removed by a rotary evaporator, and a mixed solution of ethanol and water (the volume ratio of ethanol to water is 1:2) is added to the bottle. After vigorous stirring at room temperature for 20 minutes, the mixture is filtered, and the solid is washed with 10 mL of ethanol and 10 mL of dichloromethane, respectively, and the solid is dried to obtain compound 1g. Compound 1c: Compound 1f: The molar ratio of copper sulfate: sodium ascorbate is 1:1.2:1.2:1.2, the solvent is a mixture of tetrahydrofuran and water, and the reaction is carried out at room temperature for 24 hours.

[0065] The compound 1c can be prepared by the following method:

[0066] Step 1.1, a benzoic acid derivative (1a) is subjected to an amidation condensation reaction with 2,3-diaminobenzamide and then the amino protection is removed to obtain a compound (1b).

[0067] In some embodiments, in order to promote the condensation reaction, in step 1.1, a condensation agent O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and an organic base N,N-diisopropylethylamine are also added; the molar ratio of benzoic acid derivative (1a), 2,3-diaminobenzamide, condensation agent and organic base is 1:1.05:1.05:1.5, the solvent is anhydrous DMF, the reaction temperature is room temperature, and the reaction time is 24 hours. For example, a benzoic acid derivative, an o-phenylenediamine derivative and O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate are dispersed in DMF, N,N-diisopropylethylamine is added, and the mixture is stirred at room temperature for 24 hours. After the reaction is completed, the reaction solution is slowly dripped into rapidly stirred ice water (500 mL), and a solid is precipitated. The mixture is filtered to obtain an amide intermediate. The intermediate is then dissolved in acetic acid and stirred at 120° C. for 2 hours. After the reaction is completed, the solvent acetic acid is removed by a rotary evaporator, and 30 mL of a hydrochloric acid aqueous solution (with a concentration of 4 mol / L), 40 mL of dichloromethane and 20 mL of tetrahydrofuran are continuously added to the bottle, and the mixture is stirred at room temperature for 8 hours. After the reaction is completed, the organic solvent is removed by a rotary evaporator. While the remaining reaction solution is stirred at 0° C., the pH value of the reaction solution is adjusted to 8-9 with a saturated sodium bicarbonate aqueous solution. At this time, a large amount of brown solid is precipitated, and the mixture is filtered and the filter residue is dried to obtain compound 1b.

[0068] Step 1.2, compound (1b) is subjected to amidation condensation reaction with 6-azidohexanoic acid to obtain compound (1c).

[0069] In some embodiments, in order to promote the condensation reaction, in step 1.2, a condensation agent O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, a catalyst 1-hydroxybenzotriazole, and an organic base N,N-diisopropylethylamine are also added; the molar ratio of compound (1b), 6-azidohexanoic acid, condensation agent, catalyst, and organic base is 1:1.2:1.2:0.1:5, the solvent is anhydrous DMF, the reaction temperature is room temperature, the reaction time is 12 hours, and after the reaction is completed, the reaction solution is slowly dripped into rapidly stirred ice water (500 mL), solid precipitates, and the solid is filtered and dried to obtain a crude product, which is purified by column chromatography to obtain compound 1c.

[0070] The compound 1f was prepared by the following method:

[0071] Step S1, compound (1d) and 3-bromopropyne, under alkaline conditions, undergo nucleophilic substitution reaction to form compound (1e); the alkaline conditions refer to adding a base such as potassium carbonate or sodium carbonate to the solution to make its pH value greater than 7 to promote the nucleophilic substitution reaction. As an example, the molar ratio of compound (1d), 3-bromopropyne, and inorganic base is 1:1.5:2. For example, quinoxalinone skeleton compound (1d) and potassium carbonate are dispersed in a solvent, 3-bromopropyne is added, refluxed and stirred at 73°C for 12 hours, filtered after the reaction is completed, the solid is washed with dichloromethane, and the organic solvent in the lower filtrate is removed by a rotary evaporator to obtain a crude product, a large amount of petroleum ether is added to the crude product, vigorously stirred at room temperature for 8 hours, filtered, the solid is washed with petroleum ether, and the solid is dried to obtain compound 1e. The molar ratio of compound (1d), 3-bromopropyne, and potassium carbonate is 1:1.5:2; the solvent is acetone; the reaction temperature is 73°C, and the reaction time is 12 hours.

[0072] Step S2, compound (1e) and 2,5-thiophene dicarboxaldehyde undergo condensation reaction to generate compound (1f); in order to increase the reaction rate, an organic base pyridine is also added. As an example, the molar ratio of quinoxaline skeleton compound (1e), 2,5-thiophene dicarboxaldehyde, and organic base is 1: (1-3): (1-5). For example, quinoxaline skeleton compound (1e) and 2,5-thiophene dicarboxaldehyde are dispersed in a solvent, pyridine is added, refluxed and stirred at 120°C for 6 hours, the organic solvent is removed by a rotary evaporator, 25 mL of ethanol is added to the bottle, stirred at room temperature for 20 minutes, filtered, the solid is washed with ethanol, and the solid is dried to obtain a quinoxaline thiophene derivative (compound 1f). The molar ratio of quinoxaline skeleton compound (1e), 2,5-thiophene dicarboxaldehyde, and organic base is 1: 3: 5, the solvent is acetic anhydride, the reaction temperature is 120°C, and the reaction time is 6 hours.

[0073] Step 2: Compound (1g) and compound (1j) undergo condensation reaction to obtain compound (1), thereby obtaining compound 1.

[0074] In some embodiments, an organic base tetrahydropyrrole and a catalyst indium trifluoromethanesulfonate are also added; the molar ratio of compound (1g), compound (1j), organic base and catalyst is 1: (1-3): (1-3): (1-0.5). As an example, a quinoxalinone thiophene derivative (1g), compound (1j) and catalyst indium trifluoromethanesulfonate are dispersed in a solvent, tetrahydropyrrole is added, and refluxed and stirred at 50-60°C for 2-4 hours. After the reaction is completed, the organic solvent is removed by a rotary evaporator, 15 mL of ethanol and 25 mL of water are added to the reaction bottle, stirred at room temperature for 30 minutes, filtered, and the solid is washed with 10 mL of ethanol and 10 mL of dichloromethane respectively, and the solid is dried to obtain a quinoxalinone anthraquinone derivative (Compound 1). The molar ratio of compound (1g), compound (1j), organic base and catalyst is 1: 1.5: 1.5: 0.1, and the solvent is dioxane; the reaction temperature is 55°C, and the reaction time is 2 hours.

[0075] The compound 1j was prepared by the following method:

[0076] Step A1, compound (1h) and compound (1i) undergo cyclization reaction under alkaline conditions to form compound (1j); in some implementation cases, organic base triethylamine is also added; the molar ratio of compound (1h), compound (1i) and organic base is 1:1:1.5. For example, thioisocyanate aryl derivative (1h) and carboxylic acid derivative (1i) are added to a two-necked flask equipped with a condenser, and a solvent is added under nitrogen protection, and stirred at 65°C for 12 hours. After the reaction is completed, the organic solvent is removed by a rotary evaporator, and compound (1j) is purified by column chromatography. The molar ratio of compound (1h), compound (1i) and triethylamine is 1:1:1.5, and the solvent is chloroform; the reaction temperature is 65°C, and the reaction time is 12 hours.

[0077] The following examples specifically illustrate the synthesis method of the above-mentioned biologically active photosensitizer compound:

[0078] The reagents and raw materials involved in the examples are all commercially available.

[0079] The full names of the reagent abbreviations used in the examples are as follows:

[0080] HATU: O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate

[0081] DMF: N,N-dimethylformamide

[0082] DIPEA: N,N-diisopropylethylamine

[0083] HOBT: 1-Hydroxybenzotriazole

[0084] DAPI: 4',6-diamidino-2-phenylindole

[0085] CCK-8: Cell Counting Kit-8 cell counting reagent

[0086] DMSO: dimethyl sulfoxide.

[0087] Example 1

[0088] This embodiment provides a method for preparing a bioactive photosensitizer targeting poly (ADP ribose) polymerase and androgen receptor. 1 For example, X is hydrogen and X is carbon. The specific preparation steps are as follows:

[0089]

[0090] (1) Synthesis of intermediate 1b. It includes the following steps:

[0091] Benzoic acid derivative 1a (15 mmol, 3.77 g), 2,3-diaminobenzamide (15.75 mmol, 2.38 g) and HATU (15.75 mmol, 6 g) and 30 mL of 1% ethyl acetate were added to a 250 mL dry single-necked round-bottom flask equipped with a stirrer. DMF, DIPEA (22.5mmol, 3.9mL) was added, and the mixture was stirred at room temperature for 24 hours. After the reaction, the reaction solution was slowly dripped into rapidly stirred ice water (500mL), and solid precipitated. The solid was filtered to obtain an amide intermediate. The intermediate was then dissolved in 50mL of acetic acid and stirred at 120°C for 2 hours. After the reaction, the solvent acetic acid was removed by a rotary evaporator. 30mL of hydrochloric acid aqueous solution (concentration of 4mol / L), 40mL of dichloromethane and 20mL of tetrahydrofuran were continued to be added to the bottle, and the mixture was stirred at room temperature for 8 hours. After the reaction, the organic solvent was removed by a rotary evaporator. While the remaining reaction solution was stirred at 0°C, the pH value of the reaction solution was adjusted to 8-9 with a saturated sodium bicarbonate aqueous solution. At this time, a large amount of brown solid precipitated, which was filtered and the pale yellow filter residue was dried to obtain compound 1b (3.07g, yield 77%). 1 H NMR(500MHz,DMSO-d6)δ9.34(s,1H),8.24(s,1H),8.11–8.05(m,1H),7.86(d,J=7.4Hz,1H),7.77( d,J=1.9Hz,1H),7.74(d,J=7.8Hz,1H),7.52(d,J=5.1Hz,2H),7.33(t,J=7.8Hz,1H),3.86(s,2H). 13C NMR (126MHz, DMSO-d6) δ166.4,152.3,144.8,129.4,129.1,128.9,125.7,124.9,122.8,122.1,115.4,45.4.

[0092] (2) Synthesis of intermediate 1c. It includes the following steps:

[0093] Compound 1b (4 mmol, 1.46 g), HATU (5 mmol, 1.83 g), HOBT (0.4 mmol, 52 mg) and 12 mL of DMF were added to a 250 mL dry single-necked round-bottom flask equipped with a stirrer, and then 6-azidohexanoic acid (5 mmol, 0.7 mL) and DIPEA (20 mmol, 3.5 mL) were added. The mixture was stirred at room temperature for 12 hours. After the reaction, the reaction solution was slowly dripped into rapidly stirred ice water (250 mL). Solid precipitated and was filtered and dried to obtain a crude product. The crude product was purified by column chromatography (dichloromethane / methanol = 50:1) to obtain compound 1c (990 mg, yield 61%). 1 H NMR(500MHz,DMSO-d6)δ13.43(br,1H),9.35(s,1H),8.44(t,J=5.8Hz,1H),8.13(s,1H ),8.09(d,J=7.5Hz,1H),7.87(d,J=7.3Hz,1H),7.81(s,1H),7.74(d,J=7.8Hz,1H),7. 54(t,J=7.6Hz,1H),7.42(d,J=7.6Hz,1H),7.35(t,J=7.7Hz,1H),4.38(d,J=5.9Hz,2H ),3.28(t,J=6.9Hz,2H),2.18(t,J=7.4Hz,2H),1.61–1.48(m,4H),1.39–1.27(m,2H). 13 C NMR(126MHz,DMSO-d6)δ172.6,166.7,152.4,141.9,141.3,135.8,129.96,12 9.59,126.3,125.8,123.4,122.8,115.5,51.0,42.4,35.7,28.5,26.3,25.3.

[0094] (3) Synthesis of intermediate 1j. It includes the following steps:

[0095] A stirring bar, thioisocyanate aryl derivative 1h (20 mmol, 4.56 g) and carboxylic acid derivative 1i (20 mmol, 1.78 g) were added to a 250 mL two-necked flask equipped with a condenser, and the mixture was evacuated three times. 50 mL of chloroform and triethylamine (30 mmol, 3.9 mL) were added under nitrogen protection, and stirred at 65° C. for 12 h. After the reaction was completed, the reaction solution was cooled, the organic solvent was removed by a rotary evaporator, and compound 1j (5 g, yield 83.6%) was obtained by purification by column chromatography (petroleum ether / ethyl acetate = 2:1). 1 H NMR (500MHz, DMSO-d6) δ8.34(d,J=8.2Hz,1H),8.09(d,J=1.7Hz,1H),7.93(dd,J=8.2,1.8Hz,1H),4.44(s,2H),3.30(s,3H). 19 F NMR(376MHz,DMSO-d6)δ-56.17(s,3F). 13 C NMR(101MHz,DMSO-d6)δ180.3,170.0,138.4,136.1,133.6,131.0(q,J=32.5Hz), 127.4(q,J=5.0Hz),122.2(q,J=273.7Hz),115.1,108.2(q,J=2.1Hz),54.7,33.9.

[0096] (4) Synthesis of intermediate 1e. It includes the following steps:

[0097] Compound 1d (50 mmol, 8.03 g), potassium carbonate (100 mmol, 13.8 g) and 250 mL of acetone were added to a 500 mL dry single-mouth round-bottom flask equipped with a stirrer, and 3-bromopropyne (75 mmol, 6.5 mL) was added. The mixture was refluxed and stirred at 73 ° C for 12 hours. After the reaction was completed, the reaction solution was cooled and filtered. The solid was washed with dichloromethane, the upper solid was discarded, and the organic solvent in the lower filtrate was removed by a rotary evaporator to obtain a crude product. The product was transferred to a 500 mL dry single-mouth round-bottom flask equipped with a stirrer, 450 mL of petroleum ether was added, and the mixture was vigorously stirred at room temperature for 8 hours. Filter, wash the solid with petroleum ether, and dry the solid to obtain compound 1e (8.6 g, yield 87%). 1 H NMR(400MHz, CDCl3)δ7.81(dd,J=8.0,1.1Hz,1H),7.58–7.52(m,1H),7.44(d,J=8.3H z,1H),7.39–7.33(m,1H),5.05(d,J=2.5Hz,2H),2.60(s,3H),2.28(t,J=2.5Hz,1H). 13C NMR (126MHz, CDCl3) δ158.2,154.1,132.8,131.7,129.6,129.6,123.9,114.1,73.1,31.4,21.5.

[0098] (5) Synthesis of intermediate 1f. It includes the following steps:

[0099] Compound 1e (20 mmol, 4 g), 2,5-thiophenedicarboxaldehyde (60 mmol, 8.4 g) and 30 mL of acetic anhydride were added to a 100 mL dry single-necked round-bottom flask equipped with a stirrer, and then pyridine (100 mmol, 8 mL) was added. The mixture was refluxed and stirred at 120 ° C for 6 hours. After the reaction was completed, the reaction solution was cooled, and the organic solvent was removed by a rotary evaporator. 25 mL of ethanol was added to the flask, stirred at room temperature for 20 minutes, filtered, and the solid was washed with 5 mL of ethanol. The solid was dried to obtain compound 1f (4.5 g, yield 70%). 1 HNMR (400MHz, DMSO-d6) δ9.93(s,1H),8.24(d,J=16.0Hz,1H),8.03(d,J=3.9Hz,1H),7.86(dd,J=8.0,1.2Hz,1H),7.73(d, J=3.9Hz,1H),7.71–7.61(m,2H),7.56(d,J=16.0Hz,1H),7.48–7.42(m,1H),5.13(d,J=2.3Hz,2H),3.37(t,J=2.4Hz,1H). 13 C NMR(126MHz,DMSO-d6)δ184.8,153.7,151.1,149.9,143.7,139.2,133.3, 131.9,131.6,131.3,130.1,130.0,125.6,124.8,115.6,78.4,75.8,31.9.

[0100] (6) Synthesis of intermediate 1g. It includes the following steps:

[0101] Compound 1c (0.5mmol, 203mg) and compound 1f (0.6mmol, 192mg), copper sulfate (0.6mmol, 96mg), sodium ascorbate (0.6mmol, 119mg), 12mL tetrahydrofuran and 1.5mL water were added to a 100mL dry single-mouth round-bottom flask equipped with a stirrer, and stirred at room temperature for 24 hours. After the reaction, the organic solvent in the reaction solution was removed by a rotary evaporator, 20mL ethanol and 40mL water were added to the flask, and after vigorous stirring at room temperature for 20 minutes, the mixture was filtered, and the solid was washed with 10mL ethanol and 10mL dichloromethane, respectively, and the solid was dried to obtain compound 1g (330mg, yield 91%), which can be directly used for the next step.

[0102] (7) Synthesis of the target product compound 1. The process comprises the following steps:

[0103] Compound 1g (0.5mmol, 363mg), compound 1j (0.75mmol, 225mg), indium trifluoromethanesulfonate (0.05mmol, 28mg) and 8mL 1,4-dioxane were added to a 100mL dry single-necked round-bottom flask equipped with a stirring bar, and tetrahydropyrrole (0.75mmol, 225mg) was added. The mixture was refluxed and stirred at 55°C for 2 hours. After the reaction, the organic solvent was removed by a rotary evaporator. 15mL ethanol and 25mL water were added to the reaction flask, and the mixture was stirred at room temperature for 30 minutes. The mixture was filtered, and the solid was washed with 10mL ethanol and 10mL dichloromethane, respectively. The solid was dried to obtain quinoxalinone anthraquinone derivative 1 (target product compound 1, 453mg, yield 90%). 1 H NMR (400MHz, DMSO-d6) δ13.38(s,1H),9.33(s,1H),8.43–8.34(m,2H),8.25(d,J=1.0Hz,1H) ,8.18(d,J=15.9Hz,1H),8.13–8.02(m,4H),7.89–7.76(m,4H),7.70(t,J=9.0Hz,2H),7.60– 7.47(m,4H),7.44–7.30(m,4H),5.53(s,2H),4.34(d,J=5.2Hz,2H),4.25(t,J=7.0Hz,2H),3 .66(s,3H),2.12(t,J=7.4Hz,2H),1.81–1.71(m,2H),1.59–1.48(m,2H),1.26–1.16(m,2H). 19 F NMR(376MHz,DMSO-d6)δ-60.81(s,3F).HRMS(ESI)m / z(M+H) + , calculated value (C 51 H42 F3N 12 O4S2):1007.2840; Detection value:1007.2849.

[0104] Performance Testing

[0105] (1) Absorption spectrum, fluorescence spectrum and reactive oxygen generation ability characterization of the bioactive photosensitizer obtained in the example

[0106] A 0.5 mg / mL DMSO stock solution of compound 1 was prepared, and then compound 1 was diluted to a solution with a concentration of 10 μg / mL by using a mixed solvent of DMSO / H2O in different ratios. The absorption spectrum of compound 1 was measured by a Thermo Electron-EV300 UV-visible spectrophotometer. The maximum absorption wavelength of compound 1 was at 473 nm ( Figure 3 ). The fluorescence spectrum of compound 1 was then measured by steady-state time-resolved fluorescence spectrophotometer, and the results showed that the maximum emission wavelength of compound 1 was 590 nm ( Figure 4 ). Subsequently, the reactive oxygen generation efficiency was determined by 9,10-anthracenyl-bis(methylene)dimalonic acid (ABDA) probe. The experimental results showed that compound 1 had a high reactive oxygen generation efficiency ( Figure 5 ), the horizontal axis represents the radiation time, and the vertical axis I / I0 represents the ratio of the absorbance of the mixed solution of ABDA and compound 1 after illumination to the initial absorbance.

[0107] (2) Cellular uptake and killing of prostate cancer cells

[0108] Cell uptake: Mouse prostate cancer cells MyCCAP were seeded in 3 groups of confocal microplates at a density of 10 6 / mL. Compound 1 (5 μg / ml) was added to the three groups at the same time, and the cell nuclei were stained with DAPI at 2 hours, 4 hours, and 8 hours, respectively, and the fluorescence images of the action position of compound 1 on the cells were observed by CLSM (compound 1 was excited at 495 nm, and the emission spectrum was collected at 500-550 nm). Figure 6 As shown, during the 2-hour period of administration, compound 1 was mainly concentrated in the cell cytoplasm, and during the 4-8-hour period, compound 1 was mainly concentrated in the cell nucleus.

[0109] Intracellular ROS detection: Mouse prostate cancer cells MyCCAP were seeded in 6-well plates at a density of 10 6 / mL, after attachment, compound 1 (2.5μg / ml) was added and incubated for 4 hours, then the medium was removed. After replacing with new serum-free medium, 2',7'-dichlorofluorescein diacetate (DCFH-DA) (5μM) was added and incubated for another 0.5 hours. The cells were washed three times with serum-free medium, and then blue light (60mW / cm2 , 450nm) for 5 minutes, and immediately use CLSM to observe the fluorescence image of DCFH-DA staining on the cells (DCFH-DA was excited at 495nm and the emission spectrum was collected at 500-550nm). Figure 7 As shown. It can be seen that in the ROS group, DCFH can be observed to produce obvious green fluorescence after being oxidized after illumination, indicating that compound 1 can produce obvious reactive oxygen species under intracellular illumination conditions.

[0110] Prostate cancer cell killing: MyCCAP cells were seeded in 96-well plates at a density of 5*10 4 After the cells adhered to the wall, different concentrations of compound 1 (0, 0.02 μM, 0.04 μM, 0.08 μM, 0.16 μM, 0.32 μM, 0.64 μM, 1.28 μM, 2.56 μM) were added. The illumination group was treated with blue light (60 mW / cm 2 , 450nm) for 5 minutes, the control group was not irradiated), and the culture was continued for 24 hours. Then the culture medium was removed, 100μL of fresh culture medium was added, followed by 10μL of CCK-8, and the absorbance value of each group at 450nm was measured by an enzyme marker after incubation in a 37℃ carbon dioxide incubator for 1 hour ( Figure 8 ). The results showed that IC 50 The value is 2.6μM, and the IC 50 The value is 0.028μM.

[0111] The bioactive photosensitizer of the present invention has good fluorescence efficiency and strong ability to generate active oxygen species under blue light excitation, and can be well used in image-guided photodynamic therapy.

[0112] In summary, photosensitizers convert oxygen around tumor tissue into singlet oxygen after being excited by light of a specific wavelength, thereby exerting a photodynamic therapy effect. However, tumor tissue is usually hypoxic, which limits the therapeutic effect of photosensitizers. The active photosensitizer provided by the present invention, as a PARP inhibitor at the same time, can induce cell synthetic lethality. This process can increase the oxidation state and oxygen concentration in the cell, improve the hypoxic microenvironment inside the tumor, and further enhance the photodynamic therapy effect, that is, PDT therapy and PARP inhibitors synergize.

[0113] Obviously, the above embodiments are merely examples for clear explanation and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived from these are still within the protection scope of the invention.

Claims

1. A quinoxalinone derivative targeting poly (ADP ribose) polymerase and androgen receptor, characterized in that: The structural formula of the quinoxalinone derivative is as follows: Among them, R 1 is hydrogen, methoxy, methyl or halogen, R 1 The substituent is located at least at one of the 5, 6, 7, and 8 positions of the quinoxalinone ring, and X is a carbon atom or a nitrogen atom.

2. The quinoxalinone derivative according to claim 1, characterized in that The halogen is selected from F, Cl, Br, and I.

3. A method for preparing a quinoxalinone derivative as claimed in claim 1 or 2, characterized in that: The following steps are involved: (1) Compound 1c is prepared by using compound 1b, 6-azidohexanoic acid, a condensing agent, a catalyst, and an organic base; (2) Compound 1f was prepared by using compound 1e, 2,5-thiophenedicarboxaldehyde and an organic base; (3) Compound 1g was prepared by using compound 1c, compound 1f, a copper catalyst, and a reducing agent; (4) Compound 1g, compound 1j, an organic base and a catalyst are used to prepare compound 1, i.e., a quinoxalinone derivative; wherein the structures of compound 1b, compound 1e, compound 1f, compound 1c, compound 1g, compound 1j and compound 1 are as follows:

4. The preparation method according to claim 3, characterized in that: In step (1), the molar ratio of compound 1b, 6-azidohexanoic acid, condensing agent, catalyst and organic base is 1:(1-2):(1-2):(1-0.5):(1-5).

5. The preparation method according to claim 3, characterized in that: In step (1), the catalyst is selected from 1-hydroxybenzotriazole and 4-dimethylaminopyridine; The condensing agent is selected from one or more of O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, dicyclohexylcarbodiimide, diisopropylcarbodiimide, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide; The organic base is selected from one or more of N,N-diisopropylethylamine, triethylamine and ethylenediamine.

6. The preparation method according to claim 3, characterized in that: In step (2), the molar ratio of compound 1e, 2,5-thiophenedicarboxaldehyde, and organic base is 1:(1-3):(1-5); Organic bases include pyridine and / or imidazole; Reaction conditions: reaction temperature 100-120°C, reaction time 6-12 hours.

7. The preparation method according to claim 3, characterized in that: In step (3), the molar ratio of compound 1c, compound 1f, copper catalyst and reducing agent is 1:(1-2):(1-2):(1-2); The copper catalyst is selected from one or more of copper sulfate, copper (II) bromide, copper (II) trifluoromethanesulfonate, and copper (II) acetate; The reducing agent is selected from sodium ascorbate.

8. The preparation method according to claim 3, characterized in that: In step (4), the molar ratio of compound 1g, compound 1j, organic base and catalyst is 1:(1-3):(1-3):(1-0.5); Reaction conditions: reaction temperature 50-60°C, reaction time 2-4 hours.

9. The preparation method according to claim 3, characterized in that: The organic base is selected from one or more of tetrahydropyrrole, tetrahydropyrrole, hexahydropyridine and piperazine.

10. Use of the quinoxalinone derivative according to claim 1 or 2 in the preparation of a photosensitizer for photodynamic therapy.

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