A class of 2-amino substituted pyridine structure of parp inhibitors, its pharmaceutical composition and application

Abstract

This invention belongs to the field of pharmaceutical technology, specifically relating to a class of 2-amino-substituted pyridine PARP inhibitors, their pharmaceutical compositions, and applications. The 2-amino-substituted pyridine PARP inhibitors contain an effective amount of the compound shown in formula (I) and / or its pharmaceutically acceptable derivatives: (I). The 2-amino-substituted pyridine PARP inhibitors provided by this invention exhibit higher inhibitory activity against PARP-1 enzymes and higher solubility, resulting in higher in vivo bioavailability.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a class of PARP inhibitors with 2-amino-substituted pyridine structures, their pharmaceutical compositions, and applications. Background Technology

[0002] As is well known, many anti-tumor drugs currently achieve their anti-tumor effects by damaging cellular DNA. However, this kills normal cells, and tumor cells can also develop resistance by activating intracellular DNA damage repair mechanisms. Poly(ADP-ribose) polymerase (PARP) inhibitors are the first class of anti-tumor drugs targeting DNA damage repair mechanisms. Currently, they are mainly used clinically for the treatment of BRCA-mutated, HER2-negative breast cancer or ovarian cancer, and their indications are constantly expanding.

[0003] Synthetic lethality is the main mechanism by which PARP inhibitors exert their anti-tumor effects. In normal cells, damaged DNA has two opportunities for repair: single-strand repair and double-strand repair. When a single strand of DNA is damaged, PARP enzymes, acting as molecular sensors, participate in single-strand repair. However, if single-strand repair fails, homologous recombination, a process involved in double-strand repair, will participate in DNA repair, ensuring its function. But if homologous recombination also fails, the genome will be excessively damaged, ultimately leading to apoptosis. BRCA, an early discovered tumor suppressor gene, plays a crucial role in homologous recombination repair within double-strand repair. When BRCA genes mutate in tumor cells, the DNA repair pathway relies on single-strand repair mediated by PARP enzymes. In this case, PARP inhibitors prevent DNA repair, leading to apoptosis. In contrast, in normal cells, BRCA genes function normally, still repairing DNA and allowing the cell to survive. Therefore, PARP inhibitors can be used as targeted drugs to selectively kill BRCA-mutated tumor cells. However, its mechanism of action is highly dependent on homologous recombination repair defects, and the gradual development of drug resistance during clinical use (such as compensatory activation of DNA damage repair pathways) significantly limits its clinical application. Therefore, there is an urgent need to develop PARP inhibitors with novel structures to overcome the limitations of existing inhibitors.

[0004] Regarding relevant patents published both domestically and internationally, AstraZeneca reported a novel type of selective PARP-1 inhibitor in its international patent application WO2023118085A1. The representative compound, AZD-9574, holds promise for addressing brain permeability issues and providing a new treatment option for brain metastases in breast cancer. Subsequently, related patents were reported domestically. Guangdong Dongyangguang Pharmaceutical reported a type of selective PARP-1 inhibitor with a nitrogen-containing bicyclic structure in its patent application CN119431314A. The representative compound is the compound synthesized in Example 13, whose structural core is similar to AZD-9574, differing in that a difluoromethoxy group is introduced on the benzene ring instead of a fluorine atom. Although the compounds disclosed in the above patents exhibit some selectivity for PARP1, their inhibitory activity against the PARP-1 enzyme is not high, and the compounds have very low solubility, which is detrimental to improving the bioavailability of later oral formulations.

[0005] Therefore, there is an urgent need to provide a PARP inhibitor to improve the inhibitory activity against PARP-1 enzymes and enhance its solubility and bioavailability in later oral formulations. Summary of the Invention

[0006] To address the shortcomings of the prior art, this invention provides a class of PARP inhibitors with a 2-amino-substituted pyridine structure, pharmaceutical compositions thereof, and applications. The PARP inhibitors exhibit higher inhibitory activity against PARP-1 enzymes and higher solubility, resulting in higher bioavailability of the compounds in vivo.

[0007] The specific technical solution is as follows: The first object of this invention is to provide a class of PARP inhibitors with a 2-amino-substituted pyridine structure, comprising an effective amount of the compound shown in formula (I) and / or its pharmaceutically acceptable derivatives: (I) Where X is independently selected from C, CH or N atoms; R 1 R 2 and R 3 Each is independently selected from hydrogen atoms, C1-C6 alkyl groups, or halogenated C1-C6 alkyl groups.

[0008] Furthermore, the derivative is selected from salts, solvates, hydrates, isomers, crystal forms, or prodrugs.

[0009] Furthermore, X is independently selected from C, CH, or N atoms; R 1 R 2 and R 3 Each is independently selected from hydrogen atoms or C1~C6 alkyl groups.

[0010] Furthermore, the structural formula of the PARP inhibitor is shown below: .

[0011] A second object of the present invention is to provide a pharmaceutical composition comprising a PARP inhibitor with a 2-amino-substituted pyridine structure as described above.

[0012] A third object of the present invention is to provide the use of the above-described pharmaceutical composition in the field of preparing medicaments for treating and / or preventing diseases related to PARP activity.

[0013] Furthermore, the diseases mentioned include breast cancer and ovarian cancer.

[0014] A fourth object of the present invention is to provide the use of the above-described 2-amino-substituted pyridine structure PARP inhibitor in the field of preparing pharmaceuticals for the treatment and / or prevention of diseases related to PARP activity.

[0015] Furthermore, the diseases mentioned include breast cancer and ovarian cancer.

[0016] Compared with the prior art, the beneficial effects of the present invention are as follows: The PARP inhibitors provided by this invention belong to a class of novel poly(ADP-ribose) polymerase inhibitors with a 2-amino-substituted pyridine structure. The compounds provided by this invention and / or their pharmaceutically acceptable derivatives (salts, solvates, hydrates, isomers, crystal forms, or prodrugs) can be used to prepare drugs for the treatment and / or prevention of diseases related to PARP activity (e.g., breast cancer and ovarian cancer). In vivo animal experiments have shown that the compounds of this invention have significant inhibitory effects on PARP-1 enzymes, with higher inhibitory activity; they also exhibit higher solubility in simulated intestinal fluid, resulting in higher bioavailability in later oral formulations. Attached Figure Description

[0017] Figure 1 For target compound 1 in this invention 1 H-NMR spectrum; Figure 2 For target compound 1 in this invention 13 C-NMR spectrum; Figure 3 For target compound 2 in this invention 1 H-NMR spectrum; Figure 4 For target compound 2 in this invention 13 C-NMR spectrum; Figure 5 For target compound 3 in this invention 1 H-NMR spectrum; Figure 6For target compound 3 in this invention 13 C-NMR spectrum. Detailed Implementation

[0018] The principles and features of the present invention are described below with reference to examples. The examples are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0019] Example 1 Preparation of intermediate 5:

[0020] Step a: Take a 100 mL reaction flask and weigh out 1-bromo-2,4-difluoro-3-nitrobenzene (11.85 g, 50 mmol, 1.0 eq) and alanine methyl ester hydrochloride (6.95 g, 50 mmol, 1.0 eq) sequentially. Dissolve in DMF (50 mL), and add sodium bicarbonate (25.2 g, 300 mmol, 6.0 eq) dropwise while stirring. After adding the solution, stir and react overnight in an oil bath at 30 °C. For post-treatment, extract twice with water and ethyl acetate. Dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography to give a white intermediate 1 (5.52 g), with a yield of 34%.

[0021] Step b: Take a 250 mL reaction flask and weigh out intermediate 1 (8.02 g, 28.2 mmol, 1.0 eq), iron powder (7.89 g, 141 mmol, 5.0 eq), ammonium chloride (7.54 g, 141 mmol, 5.0 eq), ethanol (100 mL), and water (30 mL). Heat the mixture in an oil bath to 50 °C and react for 4 hours. Monitor the reaction for completeness by TLC (thin-layer chromatography). Afterward, remove the heat. For post-processing, filter the reaction solution hot through diatomaceous earth, wash the filter cake with ethanol, evaporate the filtrate to dryness, and extract twice with ethyl acetate and water. Evaporate the organic phase to dryness to obtain crude intermediate 2 (6.8 g), yield 94%.

[0022] Step c: Take a 500 mL reaction flask, weigh in intermediate 2 (6.8 g, 26.3 mmol, 1.0 eq), dissolve in dichloromethane (250 mL), and add DDQ (2,3-dichloro-5,6-dicyanobenzoquinone, 7.18 g, 31.6 mmol, 1.2 eq) while stirring. Stir the reaction at room temperature for 4 hours. Monitor the reaction by TLC until complete, then stop the reaction. For post-processing, evaporate the reaction solution to dryness, slowly add sodium bicarbonate aqueous solution, stir overnight at room temperature, filter, and dry the filter cake to obtain brown solid intermediate 3 (6.5 g), yield 95%.

[0023] Step d: Take a 250 mL reaction flask and weigh out intermediate 3 (6.5 g, 25.4 mmol, 1.0 eq), Xphos (2-dicyclohexylphosphine-2′,4′,6′-triisopropylbiphenyl, 1.21 g, 2.54 mmol, 0.1 eq), Pd2(dba)3 (1.16 g, 1.27 mmol, 0.05 eq), tributyltin methanol (8.96 g, 27.9 mmol, 1.1 eq), and anhydrous dioxane (130 mL). Purge with nitrogen three times and heat in an oil bath to 80 °C overnight. Monitor the reaction by TLC until complete, then stop the reaction. For post-treatment, evaporate the reaction solution to dryness and quench with KF aqueous solution after stirring for 2 hours. The product was extracted twice with water and dichloromethane. The organic phase was dried with anhydrous sodium sulfate, and the organic phases were combined and concentrated. The crude product was purified by silica gel column chromatography to obtain gray solid intermediate 4 (2.96 g), with a yield of 56%.

[0024] Step e: Take a 250 mL reaction flask and weigh out intermediate 4 (3.6 g, 17.7 mmol, 1.0 eq) and 1,2-dibromotetrachloroethane (12.7 g, 39.1 mmol, 2.2 eq) sequentially. Dissolve in dichloromethane (80 mL), and add triethylphosphine (4.2 g, 35.6 mmol, 2.0 eq) dropwise under stirring. After the addition is complete, stir the reaction at room temperature for 6 hours. Monitor the reaction by TLC until complete, then stop the reaction. Post-processing: evaporate dichloromethane to dryness, extract twice with water and ethyl acetate, dry the organic phase with anhydrous sodium sulfate, and evaporate the organic phase to dryness to obtain the crude product. Pulp the crude product with petroleum ether, filter, and dry the filter cake to obtain key intermediate 5 (3.6 g), yield 79%.

[0025] Example 2 Preparation of target compound 1:

[0026] Step a: Synthesis of intermediate 6 (methyl 5-bromo-6-fluoropyridine-2-carboxylic acid) Take a 50 mL reaction flask, add methyl 5-bromopyridine-2-carboxylate (2.16 g, 10 mmol, 1.0 eq) and dissolve it in anhydrous acetonitrile (20 mL), add AgF2 (1.75 g, 12 mmol, 1.2 eq), remove nitrogen three times, and stir the reaction at room temperature. After reacting at room temperature for about 8 hours, add AgF2 (1.75 g, 12 mmol, 1.2 eq) and continue the reaction overnight. TLC monitoring showed that the reaction was almost complete, and the reaction was stopped. Post-processing: filter, add the filtrate to saturated ammonium chloride aqueous solution and extract three times with dichloromethane, dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure to remove the solvent, and use the crude product intermediate 6 directly in the next reaction.

[0027] Step b: Synthesis of intermediate 7 Take a 50 mL reaction flask, weigh in intermediate 6 (3.36 g, 10 mmol, 1.0 eq), dissolve in methanol (10 mL), and slowly add methylamine aqueous solution (15 mL) dropwise while stirring. After the addition is complete, stir the reaction mixture at room temperature for 24 hours. Post-processing: evaporate the reaction solution to dryness to obtain the crude product. The crude product is purified by silica gel column chromatography to obtain intermediate 7 (1.68 g), yield 69%.

[0028] Step c: Synthesis of intermediate 8 Take a 50 mL reaction flask and weigh out intermediate 7 (976 mg, 4 mmol, 1.0 eq), N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (1.36 g, 4.4 mmol, 1.1 eq), sodium carbonate (848 mg, 8 mmol, 2.0 eq), Pd(dppf)Cl2 (87 mg, 0.12 mmol, 0.06 eq), dioxane (20 mL), and water (7 mL). Purge with nitrogen three times, heat to 90 °C for 2 hours, and monitor the reaction by TLC until complete. Stop heating. For post-processing, remove the solvent under reduced pressure. Extract the reaction solution twice with water and ethyl acetate. Dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography to obtain a white solid intermediate 8 (1.07 g), yield 77%.

[0029] Step d: Synthesis of intermediate 9 Take a 25 mL reaction flask, weigh in intermediate 8 (1.07 g, 3.09 mmol, 1.0 eq), and slowly add 5 mL of 4N dioxane hydrochloride solution under ice-water bath conditions. Stir the reaction mixture overnight at room temperature. For post-processing, filter directly to the crude hydrochloride form of the product, extract twice with saturated sodium carbonate solution and ethyl acetate. Dry the organic phase with anhydrous sodium sulfate and concentrate under reduced pressure to give a white solid intermediate 9 (661 mg), yield 87%.

[0030] Step e: Synthesis of target compound 1 Take a 25 mL reaction flask and weigh out intermediate 5 (370 mg, 1.18 mmol, 1.0 eq), intermediate 9 (264 mg, 1.07 mmol, 1.1 eq), potassium iodide (35.6 mg, 0.21 mmol, 0.2 eq), DIPEA (N,N-diisopropylethylamine, 692 mg, 5.36 mmol, 5.0 eq), and acetonitrile (10 mL). Heat under reflux for 4 hours. Monitor the reaction by TLC until complete, then stop the reaction. For post-treatment, cool the reaction solution to room temperature, filter, wash the filter cake with acetonitrile and water, and dry to obtain a light-colored solid, target compound 1 (384 mg), with a yield of 74%.

[0031] Example 3 Preparation of target compound 2:

[0032] Step a: Synthesis of intermediate 10 Take a 50 mL reaction flask, weigh out intermediate 8 (3.46 g, 10 mmol, 1.0 eq) and 10% palladium on carbon (346 mg) sequentially, dissolve in ethanol (30 mL), purge with hydrogen three times, stir and heat to 70 °C. Monitor the reaction of the starting materials by TLC until complete. After post-processing, filter the reaction solution through a sintered glass funnel, and evaporate the filtrate to dryness to obtain a white foamy solid intermediate 10 (3.17 g), yield 91%.

[0033] Step b: Synthesis of intermediate 11, refer to the synthesis of intermediate 9 in Example 2.

[0034] Step c: Synthesis of target compound 2, refer to the synthesis of target compound 1 in Example 2.

[0035] Example 4 Preparation of target compound 3:

[0036] Step a: Synthesis of intermediate 12 Take a 100 mL reaction flask and weigh out the following ingredients in sequence: intermediate 7 (2.44 g, 10 mmol, 1.0 eq), N-Boc-piperazine (1.86 g, 10 mmol, 1.0 eq), Ruphos (2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl, 460 mg, 0.5 mmol, 0.05 eq), tris(dibenzylideneacetone)palladium (240 mg, 0.3 mmol, 0.03 eq), cesium carbonate (9.6 g, 3.0 eq), dioxane (30 mL), and toluene (30 mL). Purge the mixture with nitrogen three times, stir and heat to reflux for 24 hours. Monitor the reaction by TLC until it is complete, then stop heating. Post-processing: The reaction solution was cooled to room temperature, filtered to remove salt, and the filtrate was extracted twice with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to give a white solid intermediate 12 (2.65 g), with a yield of 76%.

[0037] Step b: Synthesis of intermediate 13, refer to the synthesis of intermediate 9 in Example 2.

[0038] Step c: Synthesis of target compound 3, refer to the synthesis of target compound 1 in Example 2.

[0039] test: The melting point of target compounds 1-3 obtained in Examples 2-4 of this invention was determined. 1 H-NMR, 13 The results of the C-NMR and HRMS (high-resolution mass spectrometry) tests are shown in Table 1 and... Figures 1-6 .

[0040] Table 1 Melting points of target compounds 1-3 1 H-NMR, 13 C-NMR and HRMS test results

[0041] Biological evaluation Experiment 1: In vitro inhibition of PARP-1 enzyme activity by the target compound Experimental methods: Using a commercially available PARP-1 chemiluminescence detection kit, histones were coated onto microplates and incubated overnight at 4°C. After washing three times with PBST buffer (R20863 from Shanghai Yuanye Biotechnology Co., Ltd.), the plates were blocked at room temperature for 1 hour, followed by three more washes with PBST. Serially diluted compounds 1-3, control compound 1 (AZD-9574), control compound 2 (the compound synthesized in Example 13 of patent CN119431314A), and a mixture containing PARP-1 enzyme and DNA were added, and the plates were incubated at 25°C for 30 minutes. After 30 minutes, NAD+ was added. + Initiate the reaction. After reacting at room temperature for 60 minutes, wash three times with PBST, and add poly / mono-ADP ribose-HRP antibody. After incubating at 25 °C for 1 hour, add ECL substrate A and substrate B mixed in a 1:1 volume ratio for quantitative chemiluminescence. Calculate the IC50 of the compound compared to the control group DMSO. 50 The results are shown in Table 2.

[0042] Table 2. Inhibitory activity of compounds against PARP-1 enzyme (IC50) 50 , nM)

[0043] The experimental results in Table 1 show that the target compounds 1-3 of this invention all have significant inhibitory effects on PARP-1 enzyme in vitro, and are PARP-1 inhibitors. Furthermore, the inhibitory activity of target compounds 1-3 on PARP-1 enzyme is significantly better than that of PARP-1 inhibitors reported in existing patent literature: control compound 1 (AZD-9574) and control compound 2. From the structure-activity relationship perspective, this may be due to the introduction of the amino group at position 2, which enhances the binding of the compound to PARP-1 enzyme.

[0044] Experiment 2: Determination of the solubility of the target compound in simulated intestinal fluid Experimental methods: Accurately weigh 20 mg each of target compounds 1-3, control compound 1 (AZD-9574) and control compound 2 into different vials, add artificial intestinal fluid to each vial, and equilibrate until the solubility no longer changes. HPLC is used to determine the drug concentration, and the results are shown in Table 3.

[0045] Table 3. Solubility of compounds in simulated intestinal fluid

[0046] The experimental results in Table 3 show that the solubility of the target compounds 1-3 in simulated intestinal fluid is significantly better than that of control compound 1 (AZD-9574) and control compound 2, indicating that the introduction of the amino group at position 2 enhances the solubility of the compounds, which is more conducive to improving the bioavailability of the oral formulation in vivo.

[0047] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A class of PARP inhibitors with a 2-amino-substituted pyridine structure, characterized in that, Contains an effective amount of the compound of formula (I) and / or its pharmaceutically acceptable derivatives: （I） Where X is independently selected from C, CH or N atoms; R 1 R 2 and R 3 Each is independently selected from hydrogen atoms, C1-C6 alkyl groups, or halogenated C1-C6 alkyl groups.

2. The PARP inhibitor with a 2-amino-substituted pyridine structure according to claim 1, characterized in that, The derivatives are selected from salts, solvates, hydrates, isomers, crystal forms, or prodrugs.

3. The PARP inhibitor with a 2-amino-substituted pyridine structure according to claim 1 or 2, characterized in that, X is independently selected from C, CH, or N atoms; R 1 R 2 and R 3 Each is independently selected from hydrogen atoms or C1~C6 alkyl groups.

4. The PARP inhibitor with a 2-amino-substituted pyridine structure according to claim 3, characterized in that, The structural formula of the PARP inhibitor is shown below: 。 5. A pharmaceutical composition, characterized in that, PARP inhibitors including those with a 2-amino-substituted pyridine structure as described in any one of claims 1 to 4.

6. The use of the pharmaceutical composition of claim 5 in the field of preparing medicaments for treating and / or preventing diseases related to PARP activity.

7. The application according to claim 6, characterized in that, The diseases mentioned include breast cancer and ovarian cancer.

8. The use of the 2-amino-substituted pyridine PARP inhibitor as described in any one of claims 1 to 4 in the field of preparing medicaments for the treatment and / or prevention of diseases related to PARP activity.

9. The application according to claim 8, characterized in that, The diseases mentioned include breast cancer and ovarian cancer.

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