Quinazolinone compound as well as preparation method and application thereof

Abstract

The invention discloses a quinazolinone compound with a structure shown as a formula (I) as well as a preparation method and application of the quinazolinone compound. The quinazolinone compound has an efficient inhibiting effect on PARP11, can be used for preparing a medicine for promoting interferon generation, and plays roles in resisting multiple sclerosis and chronic hepatitis B and resisting tumors. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a compound, in particular to a quinazolinone compound and a preparation method and application thereof. Background Art

[0002] Studies have shown that most PARP family members in the human body exhibit monoADP ribosyltransferase activity. The MonoPARP protein family is closely related to the development of cancer, inflammation, and neurodegenerative diseases. PARP11 is a member of the monoPARP protein family. PARP11 can regulate the protein containing β-transducin repeat sequences (β-trcp) to regulate type I interferon receptor 1 (IFNAR1) in cells, ultimately leading to a decrease in the amount of IFNAR1 on the plasma membrane, thereby inhibiting the signal transduction of type I interferon, allowing tumors to "hide" outside the immune system. Therefore, inhibiting PARP11 can restore interferon signal transduction in cells, restore the body's innate and adaptive immunity, thereby inhibiting the growth of cancer cells, enhancing the body's resistance to hepatitis B virus, regulating the immune system and reducing damage to the central nervous system.

[0003] Therefore, the development of drugs that can inhibit PARP11 is of great significance for the treatment of multiple sclerosis, chronic hepatitis B or tumors. Summary of the Invention

[0004] Purpose of the invention: The first purpose of the present invention is to provide a quinazolinone compound having good enzyme inhibitory activity against PARP11; the second purpose of the present invention is to provide a method for preparing the quinazolinone compound; the third purpose of the present invention is to provide an application of the quinazolinone compound.

[0005] Technical solution: The quinazolinone compound of the present invention has a structure of formula (I), and the quinazolinone compound includes its isomers, pharmaceutically acceptable salts or mixtures thereof;

[0006]

[0007] Wherein, X is -NH-, R 1 It is a substituted six-membered nitrogen-containing heterocycle, wherein the substituent is an alkyl group or a six-membered nitrogen-containing heterocycle.

[0008] Preferably, the substituted six-membered nitrogen-containing heterocycle is a six-membered heterocycle containing one nitrogen atom, and the substituent is on the nitrogen atom.

[0009] Preferably, the substituent on the nitrogen atom is methyl, pyrimidine or pyridine.

[0010] Preferably, the R 1 for

[0011] More preferably, the quinazolinone compound is any one of the following compounds.

[0012]

[0013] Preferably, the pharmaceutically acceptable salt is a salt of the quinazolinone compound and an acid, and the acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid or ferulic acid.

[0014] The preparation method of the quinazolinone compound of the present invention comprises a substitution reaction between compound 1-5 and compound 1-6 in an organic solvent to obtain the target compound I. The synthetic route is as follows:

[0015]

[0016] Preferably, the temperature of the substitution reaction is 70-90°C.

[0017] Preferably, the organic solvent is anhydrous DMF.

[0018] Preferably, the substitution reaction is carried out under alkaline conditions, and the alkaline environment is provided by potassium carbonate.

[0019] Preferably, the method for monitoring the progress of the substitution reaction is: detecting the reaction by thin layer chromatography, with the mobile phase being V dichloromethane:V methanol = 15:1.

[0020] Preferably, the purification method of the substitution reaction is: firstly extracting with ethyl acetate, and then purifying by silica gel column chromatography (V dichloromethane: V methanol = 15:1).

[0021] The pharmaceutical composition comprises the quinazolinone compound of the present invention and a pharmaceutically acceptable carrier. A pharmaceutically acceptable carrier can be added to prepare a common pharmaceutical preparation, such as a tablet, capsule, syrup, suspension, or injection. The preparation can also contain common pharmaceutical excipients such as flavorings, sweeteners, liquid / solid fillers, and diluents.

[0022] Use of the quinazolinone compound or pharmaceutical composition of the present invention in the preparation of PARP11 inhibitor drugs.

[0023] The invention relates to the use of the quinazolinone compound or pharmaceutical composition in the preparation of drugs for treating multiple sclerosis, chronic hepatitis B or tumors.

[0024] The tumor is lung squamous cell carcinoma, colon cancer, breast cancer or other cancers.

[0025] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: (1) the structure of the quinazolinone compounds is novel; (2) the quinazolinone compounds can effectively inhibit the PARP11 enzyme activity, and the enzyme inhibition IC 50 The optimal value is less than 0.1 μM, reaching the nanomolar concentration level; (3) This type of quinazolinone compound and its pharmaceutical composition are widely used and can be used to prepare drugs for treating multiple sclerosis, chronic hepatitis B or tumors. The drugs can exert the effect of promoting interferon production at the cellular level. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a test diagram of the promoting effect of compound 1 on interferon release. DETAILED DESCRIPTION

[0027] The technical solution of the present invention will be further described below in conjunction with embodiments.

[0028] Example 1

[0029] The quinazolinone compound of the present invention has a chemical name of 8-methyl-2-[((piperidin-4-ylmethyl)amino)methyl]-7-(prop-1-ynyl)quinazolin-4(3H)-one, and a preparation method thereof comprises the following steps:

[0030] (1) Synthesis of methyl 2-amino-3-methyl-4-(prop-1-yn-1-yl)benzoate (1-3)

[0031]

[0032] In a 100 mL three-necked flask, methyl 2-amino-3-methyl-4-bromobenzoate (1-1, 2441 mg, 10 mmol) was dissolved in anhydrous toluene (20 mL) and the atmosphere was purged with nitrogen. A solution of tributyl propargyl stannane (1-2, 4936 mg, 15 mmol) was then added via syringe, followed by rapid addition of tetrakis(triphenylphosphine)palladium (1155 mg, 1 mmol), and the atmosphere was re-purged with nitrogen. The reaction mixture was refluxed at 115°C for 2.5 h. Thin-layer chromatography (V petroleum ether:V ethyl acetate = 8:1) confirmed the reaction was complete. The reaction mixture was cooled to room temperature and concentrated. The crude product was purified by silica gel column chromatography (V petroleum ether:V ethyl acetate = 100:1) to afford 1780 mg of a yellow solid (1-3) in an 87.6% yield. ESI-MS [M+H] + 204.1.

[0033] (2) Synthesis of 2-(chloromethyl)-8-methyl-7-(prop-1-yn-1-yl)quinazolin-4(3H)-one (1-5)

[0034]

[0035] Intermediate 1-3 (1016 mg, 5 mmol) was added to a round-bottom flask and dissolved in 10 mL of 4 M hydrochloric acid (1,4-dioxane) under nitrogen. Chloroacetonitrile (1133 mg, 15 mmol) was added to the solution, and the reaction mixture was refluxed at 125°C for 5 hours. Thin-layer chromatography (V petroleum ether:V ethyl acetate = 8:1) confirmed the reaction was complete. The reaction was cooled to room temperature and adjusted to a bubble-free state with saturated sodium bicarbonate aqueous solution. The mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed sequentially with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The crude product was purified by silica gel column chromatography (V petroleum ether:V ethyl acetate = 10:1) to afford 1.1 g of a white solid (1-5) in an 89.4% yield. ESI-MS [M+H] + 247.1.

[0036] (3) Synthesis of 8-methyl-2-[((piperidin-4-ylmethyl)amino)methyl]-7-(prop-1-ynyl)quinazolin-4(3H)-one (1)

[0037]

[0038] Compound 1-5 (246.7 mg, 1.0 mmol), 4-aminomethylpiperidine (1-6, 137 mg, 1.5 mmol), and potassium carbonate (414 mg, 3.0 mmol) were added to an eggplant-shaped flask, dissolved in anhydrous DMF (5 mL), and reacted at 80°C for 4 h. The reaction was complete as determined by thin-layer chromatography (V:methylene chloride:V:methanol = 15:1). The reaction was cooled to room temperature and poured into water. Extraction was performed with ethyl acetate (5 mL x 3). The organic phases were combined, washed sequentially with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The crude product was purified by silica gel column chromatography (V:methylene chloride:V:methanol = 15:1) to yield 272 mg of a white solid (Compound 1), in an 83.8% yield. ESI-MS [M+H] + 325.2; 1 H NMR (300MHz, DMSO-d6) δ (ppm): 12.46 (s, 1H), 7.86 (d, J = 8.3Hz, 1H), 7.41 (d, J = 8.3Hz, 1H), 3.50-3.42 (m, 2H), 1. 92-2.86(m,2H),2.68-2.58(m,5H),2.35-2.32(m,1H),2.02-1.96(m,1H),1.74-1.66(m,2H),1.35-1.20(m,5H).

[0039] Example 2

[0040] The quinazolinone compound of the present invention has the chemical name: 8-methyl-2-((((1-methylpiperidin-4-yl)methyl)amino)methyl)-7-(prop-1-yn-1-yl)quinazolin-4(3H)-one, and has the following structural formula:

[0041]

[0042] Preparation method: Based on Example 1, 4-aminomethylpiperidine in step (3) was replaced with (1-methyl-4-piperidinyl)methylamine. Other conditions remained unchanged to obtain the target product, compound 2, in a yield of 84.2%. ESI-MS: [M+H] + 339.2; 1 H NMR (300MHz, DMSO-d6) δ (ppm): 12.46 (s, 1H), 7.86 (d, J = 8.3Hz, 1H), 7.41 (d, J = 8.3Hz, 1H), 3.50-3.42 (m, 2H), 1.92-2. 86(m,2H),2.68-2.58(m,5H),2.35-2.32(m,1H),2.14(s,3H),2.02-1.96(m,1H),1.74-1.66(m,2H),1.35-1.20(m,5H).

[0043] Example 3

[0044] The quinazolinone compound of the present invention has the chemical name: 8-methyl-7-(prop-1-ynyl)-2-[(((1-(2-pyrimidinyl)piperidin-4-yl)methyl)amino)methyl]quinazolin-4(3H)-one, and has the following structural formula:

[0045]

[0046] Preparation method: Based on Example 1, 4-aminomethylpiperidine in step (3) was replaced with [1-(2-pyridyl)-4-piperidyl]methylamine. Other conditions remained unchanged to obtain the target product, compound 2, in a yield of 83.6%. ESI-MS: [M+H] + 416.2; 1H NMR (300MHz, DMSO-d6) δ (ppm): 12.46 (s, 1H), 8.31 (d, J = 7.6Hz, 1H), 7.84 (d, J = 8.3Hz, 1H), 7.41 (d, J = 8.3Hz, 1H), 7.21 (d, J = 7.4Hz, 1H), 7.15-7. 08(m,2H),3.49-3.40(m,2H),1.93-2.86(m,2H),2.66-2.58(m,5H),2.3 6-2.32(m,1H),2.02-1.96(m,1H),1.74-1.66(m,2H),1.36-1.24(m,5H).

[0047] Example 4

[0048] The quinazolinone compound of the present invention has the chemical name: 8-methyl-7-(prop-1-yn-1-yl)-2-(((((1-(pyridin-2-yl)piperidin-4-yl)methyl)amino)methyl)quinazolin-4(3H)-one, and has the following structural formula:

[0049]

[0050] Preparation method: Based on Example 1, 4-aminomethylpiperidine in step (3) was replaced with (1-pyrimidin-2-ylpiperidin-4-yl)methylamine. Other conditions remained unchanged to obtain the target product, compound 2, in a yield of 85.4%. ESI-MS: [M+H] + 417.2; 1 H NMR (300MHz, DMSO-d6) δ (ppm): 12.46 (s, 1H), 8.61 (d, J = 8.2Hz, 1H), 7.84 (d, J = 8.3Hz, 1H), 7.41 (d, J = 8.3Hz, 1H), 7.29-7.21 (m, 1H), 3.49-3.40(m,2H),1.93-2.86(m,2H),2.66-2.58(m,5H),2.36-2.32(m,1H),2.00-1.96(m,1H),1.77-1.69(m,2H),1.35-1.24(m,5H).

[0051] Comparative Example 1

[0052] The compound was synthesized according to the literature A Potent and Selective PARP11 Inhibitor Suggests Coupling between Cellular Localization and Catalytic Activity (https: / / doi.org / 10.1016 / j.chembiol.2018.09.011) with the following structural formula:

[0053]

[0054] ESI-MS:[M+H] + 323.1.

[0055] Performance Testing

[0056] 1. Inhibitory activity of compounds against PARP11

[0057] Experimental materials: PARP11 Chemiluminescent Assay Kit, BPS Bioscience; DMSO, Sinopharm, Nivo, PerkinElmer.

[0058] Experimental methods:

[0059] (1) Preparation of solution and buffer:

[0060] Preparation of 10X PBS: Dissolve 720 mg of KH2PO4, 45 g of NaCl, and 5.311 g of Na2HPO4·12H2O in 500 mL of deionized water. Adjust the pH of the system to 7.4. Sterilize at 121°C for 30 min, cool, and store at 4°C until ready for use.

[0061] Preparation of 1X PBS: Dilute 10X PBS 10-fold with deionized water, i.e., add 1 part 10X PBS to 9 parts deionized water.

[0062] Wash buffer preparation: 1X PBS containing 0.05% Tween-20.

[0063] Preparation of 1X PARP buffer: (Prepare immediately before use) Dilute 10X PARP buffer 10-fold with deionized water and place on ice until ready for use.

[0064] (2) Preparation of compound working solution concentration:

[0065] According to the test requirements, the test compound was diluted with 100% DMSO to the required concentration, and then diluted 10-fold with 1X PARP buffer to prepare a 10X compound working solution.

[0066] (3) Experimental steps:

[0067] a. Thaw 5X histone mixture on ice the day before the experiment.

[0068] b. Prepare 1X histone mixture by using 1X PBS to convert 5X histone mixture into 1X histone mixture. Add 25 μL of 1X histone mixture to each well of the test plate and incubate overnight at 4°C.

[0069] c. Add 100 μL of Locking Buffer to each well of the test plate and incubate at 25°C for 90 minutes.

[0070] d. After the incubation is completed, the liquid in the test plate is dried and the plate is washed three times;

[0071] e. 2.5 μL of compound working solution was added to each well of the assay plate according to the experimental layout. Positive control wells were filled with the corresponding volume of 1X PARP buffer containing 10% DMSO, and blank wells were filled with the corresponding volume of 1X PARP buffer.

[0072] f. After the enzyme is completely dissolved, dilute the enzyme stock solution to 6 ng / μL with 1X PARP buffer;

[0073] g. Add 10 μL of enzyme solution to each well of the test plate. Add the corresponding volume of 1X PARP buffer to the blank control wells, resulting in a total enzyme volume of 60 ng per well. Note: This step must be performed on ice.

[0074] h. Add 12.5 μL of master mixture (12.5 μL master mixture includes 1.25 μL 10X PARP buffer, 1.25 μL Opti-PARP 10X Assay mixture and 10 μL water) to each well of the test plate; seal the test plate and incubate at 25°C for 60 minutes;

[0075] i. After incubation, dry the liquid in the test plate and repeat the wash three times;

[0076] j. Dilute the Streptavidin-HRP in the kit 50-fold with blocking buffer, add 25 μL per well to the test plate, and incubate at 25°C for 30 minutes;

[0077] k. After incubation, dry the liquid in the test plate and repeat the wash three times;

[0078] 1. Mix ELISA ECL Substrate A and ELISA ECL Substrate B in the kit in a 1:1 ratio, add 50 μL of the mixture to each well of the test plate, and immediately perform a luminescence assay using Nivo and read the luminescence value (RLU);

[0079] m. Enzyme rate calculation: % Enzyme Activity = (RLU (Sample) - RLU (Blank)) / (RLU (Pos.Ctrl) - RLU (Blank)) × 100%; Enzyme inhibition rate = 1 - % Enzyme Activity. The specific results are shown in Table 1 below.

[0080] Table 1. Data on the inhibitory activity of the test compounds on PARP11

[0081] Compound number <![CDATA[IC 50 (nM)]]> Example 1 89 Example 2 400 Example 3 67 Example 4 88 Comparative Example 1 203 Talazoparib 471

[0082] As shown in Table 1, all the tested compounds of the present invention showed good enzyme inhibitory activity against PARP11, and the enzyme inhibition IC 50 The values all reached the nanomolar concentration level.

[0083] 2. The promoting effect of compounds on interferon release

[0084] In the presence of the STING agonist DMXAA, RAW264.7 was tested for the induction of interferon-β levels by a PARP11 inhibitor (Compound 1). RAW264.7 cells grown to the logarithmic growth phase were plated in 96-well plates and incubated overnight at 37°C in a 5% CO2 incubator until adherence. The cells were co-treated with a dose-titrated PARP11 inhibitor and 50 μg / mL DMXAA for 24 hours and the supernatant was collected and processed by ELISA (R&D, Mouse IFN-beta DuoSet Elisa) according to the kit instructions. The results are shown in the table below. Figure 1 .

[0085] Depend on Figure 1 It can be seen that compound 1 of the present invention can significantly promote the release of interferon β, and thus can be used to treat multiple sclerosis, chronic hepatitis B or tumors.

Claims

1. A quinazolinone compound, characterized in that Having the structure of formula (I), the quinazolinone compound includes its isomers, pharmaceutically acceptable salts or mixtures thereof; Wherein, X is -NH-, R 1 It is a substituted six-membered nitrogen-containing heterocyclic ring, and the substituents are alkyl groups and six-membered nitrogen-containing heterocyclic rings.

2. The quinazolinone compound according to claim 1, wherein The substituted six-membered nitrogen-containing heterocycle is a six-membered heterocycle containing one nitrogen atom, and the substituent is on the nitrogen atom.

3. The quinazolinone compound according to claim 2, characterized in that The substituent on the nitrogen atom is H, methyl, pyrimidine or pyridine.

4. The quinazolinone compound according to claim 2, characterized in that The R 1 for 5. The quinazolinone compound according to claim 1, characterized in that The pharmaceutically acceptable salt is a salt formed by the quinazolinone compound and an acid, wherein the acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid or ferulic acid.

6. A method for preparing the quinazolinone compound according to any one of claims 1 to 5, characterized in that: Compound 1-5 and compound 1-6 undergo substitution reaction in an organic solvent to obtain the target compound I. The synthetic route is as follows:

7. The method for preparing the quinazolinone compound according to claim 3, wherein The temperature of the substitution reaction is 70-90°C.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the quinazolinone compound according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier.

9. Use of the quinazolinone compound according to any one of claims 1 to 5 or the pharmaceutical composition according to claim 8 in the preparation of PARP11 inhibitor drugs.

10. Use of the quinazolinone compound according to any one of claims 1 to 5 or the pharmaceutical composition according to claim 8 in the preparation of a drug for treating multiple sclerosis, chronic hepatitis B or tumors.

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