Quinoline derivative as well as preparation method and application thereof

Abstract

The invention belongs to the field of medicines, and relates to a quinoline derivative as well as a preparation method and application thereof. The quinoline derivative is selected from at least one compound in a structure as shown in a formula I, or a tautomer or a mixture thereof, or a prodrug thereof, or a pharmaceutically acceptable salt, a solvate or a hydrate thereof. A DNA methyltransferase inhibitory activity test, a histone deacetylase inhibitory activity test and a tumor cell proliferation inhibitory activity test on the compound provided by the invention prove that the quinoline derivative provided by the invention is a potential antitumor drug with inhibitory activity on DNA methyltransferase and histone deacetylase. The raw materials of the compound are easy to obtain, the preparation method is simple, and experiments prove that the compound has a good anti-cancer effect and has a good application prospect in the field of design, research and development of anti-tumor drugs.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and in particular relates to a quinoline derivative and a preparation method and application thereof. BACKGROUND

[0002] Epigenetic modifications, such as DNA methylation and histone deacetylation, play a key role in regulating gene expression and are closely related to the occurrence and progression of cancer. In epigenetic regulation, abnormal activation of DNA methyltransferases (DNMTs) leads to high methylation of CpG islands, while overexpression of histone deacetylases (HDACs) leads to low acetylation of histones, both of which can lead to transcriptional silencing of tumor suppressor genes and promote the occurrence and development of tumors. Therefore, dual inhibition of DNMTs and HDACs has become a promising therapeutic strategy to reverse these epigenetic abnormalities. Although several DNMT and HDAC inhibitors have been approved for the treatment of hematological malignancies, their clinical application is still limited by efficacy and safety challenges. The approved DNMT inhibitors, such as azacitidine and decitabine, are nucleoside analogs with poor specificity, leading to significant toxic side effects, limiting their use, although non-nucleoside DNMT inhibitors have shown therapeutic promise, none have been applied to the clinic. Similarly, although five HDAC inhibitors have been approved for clinical use, their limited bioavailability and poor efficacy in solid tumors remain major challenges.

[0003] DNMTs and HDACs promote the silencing of tumor suppressor genes by synergizing DNA methylation and histone deacetylation, thereby enhancing epigenetic suppression. On the one hand, the physiological synergistic mechanism of DNMTs and HDACs allows one target inhibitor to still have the potential to silence tumor suppressor genes when used alone; on the other hand, DNMT inhibitors and HDAC inhibitors have good synergistic effects when used in combination. Compared with combination therapy, dual-target drugs have synergistic effects, with advantages such as improved pharmacokinetics, reduced drug interactions, and reduced risk of drug resistance. Therefore, designing multi-target antitumor drugs targeting DNMTs and HDACs is expected to reduce drug resistance and obtain better antitumor drugs. SUMMARY

[0004] The purpose of the present application is to provide a quinoline derivative with DNMT and HDAC inhibitory activity and a preparation method and application thereof.

[0005] In order to achieve the above object, the first aspect of the present application provides a quinoline derivative selected from at least one compound represented by the structure of formula I or a tautomer thereof or a mixture thereof, or a prodrug thereof, or a pharmaceutically acceptable salt, solvate or hydrate thereof;

[0006] R1 is a substituted or unsubstituted alkyl, alkenyl, alkynyl or aryl group;

[0007] R2 is a nitrogen-containing substituent;

[0008] One of M1 and M2 is a group derived from a substituted or unsubstituted six-membered ring, and the other is a chemical bond or a hydroxyl group; preferably, the six-membered ring is a piperidine ring, a pyrimidine ring or a benzene ring, and the substituted group is an amine group;

[0009] m is an integer of 0-5, n is an integer of 0-5, and p is 0 or 1.

[0010] According to the present application, preferably, the compound represented by the structure of formula I is a compound represented by formula 1, formula 2 or formula 3.

[0011]

[0012]

[0013] n1, n2, n3 are each independently 1, 2, 3 or 4; and R3 is an amine group.

[0014] According to the present application, preferably, in the R1 substituent,

[0015] The alkyl group is a C1-C10 alkyl group, preferably a C1-C5 alkyl group;

[0016] The alkenyl group is a C2-C10 alkenyl group, preferably a C2-C5 alkenyl group;

[0017] The alkynyl group is a C2-C10 alkynyl group, preferably a C2-C5 alkynyl group;

[0018] The aryl group is a benzene group, a five-membered aromatic ring group, a six-membered aromatic ring group; the five-membered aromatic ring group is preferably a furan group, a thiophene group, an imidazole group, a pyrrole group, a pyrazole group; the six-membered aromatic ring group is preferably a pyridine group and a pyrimidine group;

[0019] The substituted group is selected from a C1-C5 alkyl group, a C1-C5 alkoxy group, a C2-C5 alkenyl group, a C2-C5 alkynyl group, a halogen, a hydroxyl group, a nitro group, a trifluoromethyl group, a trifluoromethoxy group, an amino group, an acrylamide group, a cyano group.

[0020] According to the present application, preferably, the R2 is a substituted or unsubstituted pyrrolidinyl group, a substituted or unsubstituted piperidinyl group; preferably selected from at least one of the following groups:

[0021]

[0022] According to the present application, preferably, the pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt.

[0023] Preferably, the inorganic acid salt is selected from any one of the following inorganic acids: hydrochloric acid, sulfuric acid and phosphoric acid.

[0024] Preferably, the organic acid salt is selected from any one of the following organic acids: acetic acid, trifluoroacetic acid, malonic acid, citric acid and p-toluenesulfonic acid.

[0025] According to the present application, preferably, the quinoline derivative is selected from at least one of the following compounds:

[0026]

[0027] The second aspect of the present application provides a method for preparing the quinoline derivative, comprising the following steps:

[0028] (1) reacting 5-amino-2-methoxyphenol with excess malonic acid in a solution of phosphorus oxychloride to obtain 2,4-dichloro-6-methoxy-quinolin-7-ol; and / or,

[0029] (2) introducing an alkylene segment through a nucleophilic substitution reaction; and / or,

[0030] (3) introducing a group derived from a substituted or unsubstituted six-membered ring through a nucleophilic substitution reaction; and / or,

[0031] (4) introducing an R1 group through a Suzuki coupling reaction; and / or,

[0032] (5) introducing an R2 group through a Buchwald-Hartwig coupling reaction.

[0033] According to the present application, preferably, the compound of formula 1 sequentially introduces a piperidine ring, an alkylene segment, an R1 group and an R2 group;

[0034] Preferably, the compound of formula 2 sequentially introduces an alkylene segment, an R1 group, an imine-substituted pyrimidine ring and an R2 group;

[0035] Preferably, the compound of formula 3 sequentially introduces an alkylene segment, an R1 group, an R2 group and an amine-substituted benzene ring;

[0036] Preferably, in step (1), the reaction temperature is 80-150°C, and the reaction time is 20-30h.

[0037] Preferably, in step (4), the Suzuki coupling reaction is carried out in a mixed solvent of an organic solvent and water, using a palladium catalyst, and at least one of potassium phosphate, sodium carbonate, potassium carbonate or cesium carbonate as a base; the reaction temperature is 50-150°C, and the reaction time is 1-10h.

[0038] Preferably, in step (5), the Buchwald-Hartwig coupling reaction is carried out in an organic solvent, using a palladium catalyst, and at least one of potassium phosphate, sodium carbonate, potassium carbonate or cesium carbonate as a base, the organic solvent being preferably at least one of dimethylformamide, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile; the reaction temperature is 80-200°C, and the reaction time is 20-40h.

[0039] According to a specific embodiment of the present application, the synthetic route of the compound shown in formula 1-3 is as follows:

[0040]

[0041] wherein, R1, R2 and n in compound 4, compound 5, compound 6, compound 7, compound 8, compound 9, compound 10, compound 11, compound 13, compound 14, compound 15 and compound 16 are defined the same as R1, R2 and n in formula 1, formula 2 and formula 3. Compound 12, compound 13 and compound 14 respectively undergo step d, step e and step f to obtain.

[0042] The preparation method comprises the following reaction steps:

[0043] Step a), compound 1 is reacted with excess malonic acid in a solution of phosphorus oxychloride to obtain compound 2;

[0044] Steps b and c), compound 2 undergoes a nucleophilic substitution reaction with a corresponding di-tert-butyl dicarbonate-protected halogenated piperidine, and then under trifluoroacetic acid conditions to obtain compound 3;

[0045] Step d), compound 3 undergoes a nucleophilic substitution reaction with a corresponding terminal halogenated aliphatic acid ester to obtain compound 4;

[0046] Step e), compound 4 and 7 undergo a Suzuki coupling reaction with a corresponding boronic acid derivative carrying a R1 group to obtain compound 5 and 8;

[0047] Step f), compound 5 and 10 undergo a Buchwald-Hartwig coupling reaction with a corresponding amino derivative carrying a R2 group to obtain compound 6 and 11;

[0048] Step g), reacting compound 6 with hydroxylamine or hydroxylamine hydrochloride in a basic solution to obtain a compound of formula 1 ;

[0049] Step a1), subjecting compound 2 to a nucleophilic substitution reaction with a corresponding terminal halogenated di-tert-butyl dicarbonate protected alkane amino compound to obtain compound 7;

[0050] Step a2), subjecting 2-chloropyrimidine-5-carboxylic acid methyl ester to a nucleophilic substitution reaction with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine to obtain compound 2a;

[0051] Step h), deprotection of compound 8 with trifluoroacetic acid to obtain compound 9;

[0052] Step i), subjecting compound 9 to a nucleophilic substitution reaction with compound 2a to obtain compound 10;

[0053] Step j), reacting compound 11 in an acidic solution to obtain a compound of formula 2.

[0054] Step k), hydrolysis of compound 14 under basic conditions to obtain compound 15;

[0055] Step I), condensation of compound 15 with a diamino aromatic compound to obtain compound 16;

[0056] Step m), deprotection of compound 15 under acidic conditions to obtain a compound of formula 3;

[0057] In step a), the reaction temperature is 100 °C and the reaction time is 24 h;

[0058] Steps b, c), the reaction is first carried out under basic conditions, the base is preferably at least one selected from the group consisting of triethylamine, diisopropylethylamine, potassium phosphate, sodium carbonate, potassium carbonate and cesium carbonate; the reaction is carried out in an organic solvent, the organic solvent is preferably at least one selected from the group consisting of dimethylformamide, dimethylacetamide, N-methylpyrrolidone, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile. Subsequently, the reaction is carried out under acidic conditions, the acid is preferably at least one selected from the group consisting of trifluoroacetic acid, trichloroacetic acid, malonic acid, benzene sulfonic acid, methyl sulfonic acid, ethyl sulfonic acid, hydrochloric acid and sulfuric acid; the reaction is carried out in an organic solvent, the organic solvent is preferably at least one selected from the group consisting of ethyl acetate, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile;

[0059] Step d), the reaction is carried out under basic conditions, the base is preferably at least one selected from the group consisting of triethylamine, tert-butylamine, diisopropylethylamine, potassium phosphate, sodium carbonate, potassium carbonate and cesium carbonate; the reaction is carried out in an organic solvent, the organic solvent is preferably at least one selected from the group consisting of dimethylformamide, dimethylacetamide, N-methylpyrrolidone, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile;

[0060] Step e), the boronic acid derivative carrying the R1group is pinacol boronic acid derivative; the Suzuki coupling reaction is carried out in a mixed solvent of organic solvent and water, using palladium catalyst, and using at least one of potassium phosphate, sodium carbonate, potassium carbonate or cesium carbonate as base;

[0061] Step f), the amino derivative carrying the R2group undergoes Buchwald-Hartwig coupling reaction, the base is selected from at least one of potassium phosphate, sodium carbonate, potassium carbonate or cesium carbonate; using palladium catalyst, and using at least one of potassium phosphate, sodium carbonate, potassium carbonate or cesium carbonate as base, the reaction is carried out in organic solvent, which is preferably selected from at least one of dimethylformamide, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile;

[0062] Step g), the base is selected from at least one of sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide, sodium tert-butoxide and potassium tert-butoxide; the reaction is carried out in aqueous solution or organic solvent, which is preferably selected from at least one of dimethylformamide, chloroform, dichloromethane, methanol, ethanol, acetone, tetrahydrofuran and acetonitrile; the molar ratio of the compound represented by formula 1 to hydroxylamine or hydroxylamine hydrochloride, base is 1:1-10:1-20; the reaction temperature is 20-80℃, and the reaction time is 0.2-10h;

[0063] Step a1), the reaction is carried out under basic conditions, the base is preferably selected from at least one of triethylamine, tert-butylamine, diisopropylethylamine, sodium carbonate, potassium carbonate and cesium carbonate; the reaction is carried out in organic solvent, which is preferably selected from at least one of dimethylformamide, dimethylacetamide, N-methylpyrrolidone, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile;

[0064] Step a2), the reaction is carried out under basic conditions, the base is preferably selected from at least one of triethylamine, tert-butylamine, diisopropylethylamine, potassium phosphate, sodium carbonate, potassium carbonate and cesium carbonate; the reaction is carried out in organic solvent, which is preferably selected from at least one of dimethylformamide, dimethylacetamide, N-methylpyrrolidone, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile;

[0065] Step h), the reaction is carried out under acidic conditions, the acid is preferably selected from at least one of trifluoroacetic acid, trichloroacetic acid, malonic acid, benzene sulfonic acid, methyl sulfonic acid, ethyl sulfonic acid, hydrochloric acid and sulfuric acid; the reaction is carried out in organic solvent, which is preferably selected from at least one of ethyl acetate, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile;

[0066] Step i), the reaction is carried out under basic conditions, the base is preferably at least one of triethylamine, tert-butylamine, diisopropylethylamine, potassium phosphate, sodium carbonate, potassium carbonate and cesium carbonate; the reaction is carried out in an organic solvent, the organic solvent is preferably at least one of dimethylformamide, dimethylacetamide, N-methylpyrrolidone, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile, and the reaction temperature is 20-120°C;

[0067] Step j), the reaction is carried out under acidic conditions, the acid is preferably at least one of an ethanolic hydrochloric acid solution, a methanolic hydrochloric acid solution, an isopropanolic hydrochloric acid solution, an ethyl acetate hydrochloric acid solution, an aqueous hydrochloric acid solution, an aqueous trifluoroacetic acid solution;

[0068] Step k), the reaction is carried out under basic conditions, the base is at least one of sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide, sodium tert-butoxide and potassium tert-butoxide, the reaction is carried out in a mixed solvent of an organic solvent and water, the organic solvent is preferably at least one of methanol, ethanol, isopropanol, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile;

[0069] Step I), the reaction is carried out under basic conditions, the base is preferably at least one of triethylamine, tert-butylamine, diisopropylethylamine, potassium phosphate, sodium carbonate, potassium carbonate and cesium carbonate; the reaction is carried out in an organic solvent, the organic solvent is preferably at least one of dimethylformamide, dimethylacetamide, N-methylpyrrolidone, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile;

[0070] Step m), the reaction is carried out under acidic conditions, the acid is preferably at least one of trifluoroacetic acid, trichloroacetic acid, malonic acid, benzene sulfonic acid, methyl sulfonic acid, ethyl sulfonic acid, hydrochloric acid and sulfuric acid.

[0071] The third aspect of the present application provides a pharmaceutical composition comprising the quinoline derivative as an active ingredient and a pharmaceutically acceptable adjuvant;

[0072] The preparation form of the pharmaceutical composition is preferably selected from injection solution, tablet, powder, granule, capsule, oral solution, ointment, cream.

[0073] The fourth aspect of the present application provides the use of the quinoline derivative and / or the pharmaceutical composition in at least one of the following aspects:

[0074] 1) preparation of DNA methyltransferase and histone deacetylase inhibitors;

[0075] 2) preparation of eukaryotic tumor cell proliferation inhibitors;

[0076] 3) preparation of drugs for preventing and / or treating tumors.

[0077] According to the present application, preferably, the DNA methyltransferase comprises the subtypes in mammalian cells: DNMT1, DNMT3A, DNMT3B.

[0078] According to the present application, preferably, the histone deacetylase comprises the subtypes in mammalian cells: HDAC1, HDAC2, HDAC3, HDAC8, HDAC4, HDAC5, HDAC7, HDAC9, HDAC6, HDAC10, HDAC11.

[0079] According to the present application, preferably, the eukaryote is a mammal; the tumor cell is a cancer cell; the cancer cell is preferably selected from leukemia cancer cell, breast cancer cell, liver cancer cell, pancreatic cancer cell, lung cancer cell, brain cancer cell, ovarian cancer cell, uterine cancer cell, testicular cancer cell, skin cancer cell, gastric cancer cell, nasopharyngeal cancer cell, colon cancer cell, bladder cancer cell or rectal cancer cell; more preferably, the cancer cell is human chronic myelogenous leukemia cell and / or human histiocytic lymphoma cell.

[0080] According to the present application, preferably, the leukemia cancer cell is preferably human chronic myelogenous leukemia cell line K562, the lymphoma cell is preferably human histiocytic lymphoma cell U937, the lung cancer cell is preferably human lung cancer cell NCI-H520, the human brain glioma cell is preferably U251, the melanoma cancer cell is preferably A375, the glioblastoma cell is preferably human glioblastoma cell A172 and human brain astrocytic glioma cell U-118MG, the cervical cancer cell is preferably human cervical cancer cell line Hela, the nasopharyngeal cancer cell is preferably nasopharyngeal cancer cell line CNE-2, the liver cancer cell is preferably human liver cancer cell line HepG2, the breast cancer cell is preferably human breast cancer cell MCF-7 and MDA-MB-231.

[0081] According to the present application, preferably, the tumor is cancer; the cancer is leukemia, lymphoma, lung cancer, melanoma, glioblastoma, cervical cancer, nasopharyngeal cancer, liver cancer, breast cancer, brain cancer, pancreatic cancer, ovarian cancer, uterine cancer, testicular cancer, skin cancer, gastric cancer, colon cancer, bladder cancer or rectal cancer.

[0082] The compound provided by the present application is tested by DNA methyltransferase inhibition activity test, histone deacetylase inhibition activity test and tumor cell proliferation inhibition activity test, which proves that the quinoline derivative of the present application is a potential anti-tumor drug with inhibition activity on DNA methyltransferase and histone deacetylase. The compound provided by the present application has easy raw material, simple preparation method and good anti-cancer effect proved by experiments, and has good application prospect in the field of anti-tumor drug design and development.

[0083] Other features and advantages of the present application will be described in detail in the following detailed description section. DETAILED DESCRIPTION

[0084] Preferred embodiments of the present application will be described in more detail below. Although the following describes preferred embodiments of the present application, it should be understood that the present application can be implemented in various forms and should not be limited by the embodiments set forth herein.

[0085] Example 1 Preparation of compound Y1

[0086] Example 1A Compound 2 2,4-dichloro-6-methoxy-quinolin-7-ol

[0087] Into a round bottom flask, 5-amino-2-methoxyphenol (1 eq.) and malonic acid (2 eq.) were added, followed by the addition of phosphorous oxychloride. The reaction was stirred at room temperature for 4 hours under nitrogen protection, and then heated to 90 °C overnight. After removing most of the phosphorous oxychloride by concentration under reduced pressure, the resulting oil was slowly added dropwise into ice water, and 6M NaOH was added to adjust the pH to neutral. The crude product was then obtained by ethyl acetate extraction, and compound 2 was obtained by column chromatography.

[0088] Compound characterization: 1H NMR (400 MHz, Chloroform-d) δ 7.46 (s, 1H), 7.40 (s, 1H), 7.36 (s, 1H), 4.10 (s, 3H). 13 C NMR (101 MHz, CDC13) δ 150.46, 148.55, 147.91, 145.51, 142.26, 120.51, 119.66, 111.07, 101.62, 56.41.

[0089] Example 1B Compound 3 2-[4-({[(2,4-dichloro-6-methoxyquinolin-7-yl)oxy]methyl}piperidin-1-yl]acetic acid ethyl ester

[0090] Into a round bottom flask, the compound obtained in Example 1A and 1-BOC-4- bromomethylpiperidine (1.2 eq.) were added, followed by the addition of DMF to dissolve the raw materials, and then the addition of cesium carbonate (1.5 eq.) as a base. The reaction was stirred at room temperature, and TLC was used to monitor the reaction until the raw materials were completely reacted. After the reaction was completed, the reaction solution was poured into water, and the crude product was obtained by filtration after standing. The crude product was dissolved in DCM, and then trifluoroacetic acid was added. TLC was used to monitor the reaction until the raw materials were completely reacted, and then the pH was adjusted by adding a base. The crude product was separated by column chromatography to obtain compound 3.

[0091] Example 1C Compound 4 2-(4-((2,4-dichloro-6-methoxyquinolin-7-yl)oxy)methyl)piperidin-1-yl)acetic acid ethyl ester (n = 1)

[0092] The compound from Example IB was added to a flask along with ethyl bromoacetate (1.2 eq.). After the starting material was dissolved in DMF, cesium carbonate (1.5 eq.) was added as a base. The reaction was stirred at room temperature until the starting material was consumed by TLC. After the reaction was complete, the reaction was poured into water and allowed to stand. The crude product was collected by filtration. The compound 4 was isolated by column chromatography.

[0093] Example ID Compound 5 Ethyl 2-[4-({[4-chloro-6-methoxy-2-(5-methylfuran-2-yl)quinolin-7- yl]oxy}methyl)piperidin-1-yl]acetate (R1 is 2-methylfuran)

[0094] This reaction was accomplished by a Suzuki reaction. In general, the compound from Example IB (1 eq.) was dissolved in a mixture of dioxane and water. Sodium carbonate (2 eq.), palladium catalyst tetrakis(triphenylphosphine)palladium (0.1 eq.), and the corresponding pinacol boronate derivative (2-methylfuran-5- boronic acid pinacol ester) (2 eq.) were added. The reaction was stirred at 80 °C under a nitrogen atmosphere for 2 h until the starting material was consumed by TLC. The reaction was extracted with ethyl acetate and the compound 5 was isolated by column chromatography.

[0095] Example IF Compound 6 Ethyl 2-[4-({[6-methoxy-2-(5-methylfuran-2-yl)-4-((1-methylpiperidin-4- yl)amino)quinolin-7-yl]oxy}methyl)piperidin-1-yl]acetate (R2 is N-methylpiperidine)

[0096] This reaction was accomplished by a Buchwald-Hartwig reaction. The compound from Example ID (1 eq.) and 1-methyl-4-aminopiperidine (10 eq.) were dissolved in dioxane. Pd2(dba)3 (0.2 eq.) was used as the catalyst and BINAP (0.4 eq.) was used as the ligand. Cesium carbonate (5 eq.) was added and the reaction was stirred at 100 °C under a nitrogen atmosphere for about 24 h. The reaction was extracted with ethyl acetate and the compound 6 was isolated by column chromatography.

[0097] Example IG Compound Y1 N-hydroxy-2-[4-({[6-methoxy-2-(5-methylfuran-2-yl)-4-((1-methylpiperidin-4- yl)amino)quinolin-7-yl]oxy}methyl)piperidin-1-yl]acetamide

[0098] The compound obtained in Example 1F (1 eq.) was first added to a round bottom flask, and an appropriate amount of methanol or dichloromethane was added to fully dissolve it. A hydroxylamine aqueous solution (15.17 M, 5 eq.) and a sodium methoxide methanol solution (5 M, 10 eq.) were sequentially added, and the reaction was stirred at room temperature. TLC monitoring was performed until the starting material was completely reacted. The reaction solution was removed under reduced pressure, and an appropriate amount of water was added to dissolve the solid. A 3 M hydrochloric acid was added dropwise to adjust the pH to neutral, and a white precipitate was generated at this time. Filtration and washing of the filter cake with diethyl ether, n-hexane, and the like yielded the target compound Y1.

[0099] Compound characterization: 1 H NMR (400 MHz, DMSO-d6) δ 7.57 (s, 1H), 7.18 (s, 1H), 7.04 (d, J = 3.2 Hz, 1H), 6.75 (d, J = 8.6 Hz, 2H), 6.26 (d, J = 3.1 Hz, 1H), 3.94-3.92 (m, 2H), 3.92 (s, 3H), 3.77-3.65 (m, 2H), 3.28 (s, 1H), 3.11 (d, J = 11.1 Hz, 2H), 2.88 (s, 2H), 2.85 (s, 1H), 2.45 (s, 3H), 2.39 (s, 3H), 2.07 (q, J = 8.6, 4.8 Hz, 4H), 1.88-1.74 (m, 5H), 1.46-1.32 (m, 2H), 1.23 (s, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 166.42, 152.94, 151.57, 149.11, 148.15, 147.23, 144.98, 112.32, 110.03, 108.99, 108.83, 102.27, 93.02, 72.96, 60.06, 56.77, 54.09, 53.41, 48.60, 44.97, 35.20, 30.58, 28.90, 21.23, 14.56, 14.12. HRMS (ESI): calcd for m / z C 29 H 39 N5O5, [M+H]+538.3024, found: 538.3017.

[0100] Preparation of compound Y2 in Example 2

[0101] Compound preparation method according to Example 1. Compound characterization: 1H NMR (400 MHz, DMSO-d6) δ 10.54 (s, 1H), 7.62 (s, 1H), 7.21 (s, 1H), 7.08 (d, J = 3.2 Hz, 1H), 6.95 - 6.85 (m, 1H), 6.75 (s, 1H), 6.27 (d, J = 3.2 Hz, 1H), 3.95 (s, 2H), 3.92 (s, 3H), 3.84 - 3.74 (m, 2H), 3.23 (d, J = 11.3 Hz, 2H), 2.99 (d, J = 10.8 Hz, 2H), 2.77 (t, J = 10.9 Hz, 2H), 2.66 (d, J = 7.6 Hz, 2H), 2.54 (s, 2H), 2.39 (s, 3H), 2.24 (t, J = 7.2 Hz, 2H), 2.19 - 2.16 (m, 1H), 2.10 - 2.07 (m, 2H), 1.99 - 1.91 (m, 2H), 1.82 - 1.79 (m, 2H), 1.43 - 1.33 (m, 2H), 1.21 (s, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 168.08, 153.07, 152.69, 151.52, 149.19, 148.13, 147.02, 144.71, 112.27, 110.32, 108.90, 108.74, 102.28, 92.99, 72.65, 56.77, 54.13, 53.61, 52.67, 48.10, 44.22, 34.97, 30.12, 29.92, 29.49, 28.31, 22.57, 14.14. HRMS (ESI): calcd for m / z C 30 H 41 N5O5, [M+H]+ 552.3180, found: 552.3175.

[0102] Preparation of compound Y3 of Example 3

[0103] The compounds were prepared according to the procedure of Example 1. The compounds were characterized: 1H NMR (400 MHz, DMSO-d6) δ 10.38 (s, 1H), 7.52 (s, 1H), 7.13 (s, 1H), 6.98 (d, J = 3.2 Hz, 1H), 6.70 (s, 1H), 6.57 (d, J = 7.7 Hz, 1H), 6.23 (s, 1H), 3.96 - 3.87 (m, 2H), 3.40 (s, 3H), 2.93 - 2.80 (m, 2H), 2.38 (s, 3H), 2.25 - 2.22 (m, 4H), 2.08 (t, J = 11.7 Hz, 3H), 2.04 - 1.93 (m, 5H), 1.91 - 1.85 (m, 3H), 1.78 - 1.75 (m, 2H), 1.72 - 1.68 (m, 2H), 1.67 - 1.62 (m, 2H), 1.33 (t, J = 11.9 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 169.56, 166.55, 153.49, 152.63, 151.34, 148.98, 147.98, 147.66, 145.56, 112.39, 109.53, 109.32, 108.73, 102.14, 92.92, 72.93, 58.12, 56.69, 55.04, 53.36, 49.61, 46.52, 35.74, 31.81, 30.82, 29.14, 25.55, 23.15, 19.94, 14.11. HRMS (ESI): calcd for m / z C 31 H 43 N5O5, [M+H] + 566.3337, found: 566.3340.

[0104] Preparation of compound Y4 of Example 4

[0105] Compound 7 tert-butyl (3-((2,4-dichloro-6-methoxyquinolin-7- yl)oxy)propyl)carbamate of Example 4A

[0106] Into a round bottom flask was added compound 2 from Example 1A and tert-butyl (3-((2,4-dichloro-6-methoxyquinolin-7-yl)oxy)propyl)carbamate (1.2 eq.), the starting material was dissolved in DMF, then cesium carbonate (1.5 eq.) was added as a base, the reaction was stirred at room temperature, TLC was used to monitor the reaction until the starting material was completely consumed. After the reaction was completed, the reaction solution was poured into water, extracted with ethyl acetate, and the crude product was separated by column chromatography to obtain compound 7.

[0107] Example 4B Compound 8 tert-Butyl (3-((2,4-dichloro-6-methoxyquinolin-7-yl)oxy)propyl)carbamate (R1is 2-methylfuran)

[0108] Reference Example 1D

[0109] Example 4C Compound 9 3-[(4-chloro-2-(5-methylfuran-2-yl)-6-methoxyquinolin-7- yl)oxy]propan-1 -amine

[0110] The compound 8 from Example 4B (1 eq.) was added to a round bottom flask, dissolved in dichloromethane, followed by trifluoroacetic acid solution, stirred at room temperature, monitored by TLC until the starting material was consumed. The reaction mixture was adjusted to neutral or weak basic by adding appropriate amount of water, extracted by dichloromethane, the crude product was purified by column chromatography to give compound 9.

[0111] Example 4D Compound 2a 2-chloro-N-[(tetrahydro-2H-pyran-2-yl)oxy]pyrimidine-5- carboxamide

[0112] Methyl 2-chloropyrimidine-5-carboxylate (1 eq.) and O-(tetrahydro-2H-pyran-2- yl)hydroxylamine (1.5 eq.) were added to a round bottom flask, dissolved in DMF, followed by HOBt (1.5 eq.), EDCI (1.5 eq.), DIPEA (2 eq.), stirred at room temperature, monitored by TLC until the starting material was consumed. The reaction mixture was extracted by ethyl acetate, the crude product was purified by column chromatography to give compound 2a.

[0113] Example 4E Compound 10 2-[(3-((4-chloro-2-(5-methylfuran-2-yl)-6-methoxyquinolin-7- yl)oxy)propyl)amino]-N-[(tetrahydro-2H-pyran-2-yl)oxy]pyrimidine-5-carboxamide

[0114] Compound 9 from Example 4C (1 eq.) and compound 10 from Example 2D (1.2 eq.) were added to a round bottom flask, dissolved in CH3CN, followed by potassium carbonate (2.0 eq.), stirred at 80 °C, monitored by TLC until the starting material was consumed. The reaction mixture was extracted by ethyl acetate, the crude product was purified by column chromatography to give compound 10.

[0115] Example 4F Compound 11 2-[(3-((6-methoxy-2-(5-methylfuran-2-yl)-4-((1- methylpiperidin-4-yl)amino)quinolin-7-yl)oxy)propyl)amino]-N-[(tetrahydro-2H-pyran-2- yl)oxy]pyrimidine-5-carboxamide

[0116] Reference Example 1F

[0117] Example 4G Compound Y4 N-hydroxy-2-[(3-((6-methoxy-2-(5-methylfuran-2-yl)-4-((1- methylpiperidin-4-yl)amino)quinolin-7-yl)oxy)propyl)amino]pyrimidine-5-carboxamide

[0118] To a solution of Example 4F compound 11 (1 eq.) was added 2M HC1 / EA. The mixture was stirred at room temperature, TLC monitored until the starting material was completely reacted. The reaction solution was concentrated, dissolved in a small amount of methanol. Crystallization, washed with ether, product Y4 was obtained.

[0119] Compound characterization: 1 H NMR (400 MHz, Methanol-d4) δ 8.79 (s, 2H), 7.82 (s, 1H), 7.71 (s, 1H), 7.52 (s, 1H), 7.14 (s, 1H), 6.46 (s, 1H), 4.37 (t, J = 5.5 Hz, 3H), 4.04 (s, 3H), 3.83 (t, J = 6.3 Hz, 2H), 3.71 (d, J = 11.2 Hz, 2H), 3.44 - 3.36 (m, 2H), 2.98 (s, 3H), 2.53 (s, 3H), 2.43 - 2.36 (m, 2H), 2.35 - 2.23 (m, 4H), 1.33 - 1.30 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 163.14, 157.37, 153.97, 153.42, 149.30, 144.48, 139.86, 135.24, 130.12, 117.39, 115.29, 110.64, 110.52, 103.52, 101.58, 67.19, 57.33, 52.94, 48.31, 45.91, 43.16, 28.79, 14.24, 8.95. HRMS (ESI): calcd for m / z C 29 H 35 N7O5, [M+H] + 562.2772, found: 562.2767.

[0120] Preparation of Example 5 Compound Y5

[0121] Compound preparation method refers to Example 4. Compound characterization: 1H NMR (400 MHz, DMSO-d6) δ 13.74 (s, 1H), 11.06 (s, 1H), 8.67 (d, J = 8.5 Hz, 3H), 8.23 (s, 1H), 8.10 (d, J = 3.5 Hz, 1H), 7.96 (s, 1H), 7.92 (s, 1H), 6.99 (s, 1H), 6.50 (d, J = 3.3 Hz, 1H), 4.45 - 4.20 (m, 3H), 4.11 (t, J = 6.5 Hz, 2H), 3.96 (s, 3H), 3.54 (s, 1H), 3.43 (t, J = 7.0 Hz, 2H), 3.33 - 3.24 (m, 2H), 2.77 (s, 1H), 2.75 (s, 1H), 2.47 (s, 3H), 2.31 - 2.22 (m, 2H), 2.17 - 2.14 (m, 2H), 1.88 (p, J = 6.8 Hz, 2H), 1.73 (p, J = 7.0 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 162.05, 161.89, 157.44, 157.21, 153.80, 153.33, 149.18, 144.45, 139.77, 135.28, 117.56, 115.11, 110.55, 110.42, 103.52, 101.55, 92.30, 68.95, 57.32, 52.86, 48.31, 43.14, 40.93, 28.78, 26.17, 25.74, 14.20. HRMS (ESI): calcd for m / z C 30 H 37 N7O5, [M+H] + 576.2929, found: 576.2921.

[0122] Preparation of compound Y6 of Example 6

[0123] The compound was prepared according to the procedure described in Example 4. Characterization of the compound: White solid, 15% yield. 1HNMR (400 MHz, DMSO-d6) δ 13.75 (s, 1H), 11.08 (s, 1H), 8.67 (d, J = 6.6 Hz, 3H), 8.19 (s, 1H), 8.10 (d, J = 3.4 Hz, 1H), 7.96 (s, 1H), 7.93 (s, 1H), 6.99 (d, J = 5.8 Hz, 1H), 6.50 (d, J = 3.3 Hz, 1H), 4.49 - 4.18 (m, 3H), 4.08 (t, J = 6.5 Hz, 2H), 3.96 (s, 3H), 3.51 (s, 1H), 3.37 (t, J = 6.9 Hz, 2H), 3.28 (d, J = 11.7 Hz, 2H), 2.76 (s, 1H), 2.75 (s, 1H), 2.46 (s, 3H), 2.26 (q, J = 11.3 Hz, 2H), 2.17 - 2.14 (m, 2H), 1.84 (p, J = 6.9 Hz, 2H), 1.61 (q, J = 7.3 Hz, 2H), 1.52 - 1.45 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 161.98, 161.66, 157.39, 157.19, 153.78, 153.29, 149.14, 144.44, 139.72, 135.26, 117.56, 114.97, 110.49, 110.43, 103.41, 101.47, 92.26, 69.13, 57.28, 52.84, 48.28, 43.12, 41.14, 28.84, 28.77, 28.41, 23.33, 14.22. HRMS (ESI): calcd for m / z C 31 H 39 N7O5, [M+H] + 590.3085, found: 590.3076.

[0124] Preparation of compound Y7 of Example 7

[0125] Preparation of compound 12 5-[(2,4-dichloro-6-methoxyquinolin-7-yl)oxy]pentanoic acid ethyl ester

[0126] Preparation of compound 13 5-[(4-chloro-6-methoxy-2-(5-methylfuran-2-yl)quinolin-7-yl)oxy]pentanoic acid ethyl ester

[0127] Preparation of compound 14

[0128] Example 7C Compound 14 5-[(6-methoxy-2-(5-methylfuran-2-yl)-4-((1-methylpiperidin-4- yl)amino)quinolin-7-yl)oxy]pentanoic acid ethyl ester

[0129] Method of compound preparation Reference Example 1F

[0130] Example 7D Compound 15 5-[(6-methoxy-2-(5-methylfuran-2-yl)-4-((1-methylpiperidin-4- yl)amino)quinolin-7-yl)oxy]pentanoic acid

[0131] Example 7D Compound 14 (1 eq.) and 5M NaOH solution in MeOH / H20 were reacted at room temperature, monitored by TLC until the starting material was completely reacted. The pH of the mixture was adjusted to neutral or weakly acidic with 1M HCI (aq.) and concentrated under reduced pressure. Then, the mixture was cooled to 4°C and filtered to obtain crude product 15.

[0132] Example 7E Compound 16 N-tert-butoxycarbonyl-2-[5-((6-methoxy-2-(5-methylfuran-2-yl)-4-((1- methylpiperidin-4-yl)amino)quinolin-7-yl)oxy)pentanamido]aniline

[0133] Example 7D Compound 15 (1 eq.) and tert-butyl (2-aminophenyl)carbamate (1.1 eq.), HATU (1.5 eq.), NEt3(2 eq.) were charged in a round bottom flask, dissolved and stirred at room temperature, monitored by TLC until the starting material was completely reacted. Compound 16 was obtained after column chromatography of the crude.

[0134] Example 7F Compound Y7 N-(2-aminophenyl)-5-[(6-methoxy-2-(5-methylfuran-2-yl)-4-((1- methylpiperidin-4-yl)amino)quinolin-7-yl)oxy]pentanamide

[0135] Example 7F Compound 16 (1 eq.) was dissolved in dichloromethane in a round bottom flask, followed by the addition of trifluoroacetic acid, stirred at room temperature, monitored by TLC until the starting material was completely reacted. The pH of the mixture was adjusted to weakly basic with 2M NaOH (aq.) and extracted with DCM. Compound Y7 was obtained after column chromatography of the crude.

[0136] Compound characterization: 1H NMR (600 MHz, DMSO-d6) δ 9.64 (s, 2H), 9.25 (s, 1H), 8.52 (d, J = 7.7 Hz, 1H), 7.88 (s, 1H), 7.81 (d, J = 3.5 Hz, 1H), 7.70 (s, 1H), 7.03 (s, 1H), 6.89 (t, J = 7.4 Hz, 1H), 6.73 (d, J = 7.8 Hz, 1H), 6.56-6.52 (m, 2H), 4.25 (d, J = 9.8 Hz, 1H), 4.17 (t, J = 6.5 Hz, 2H), 3.97 (s, 3H), 3.59 (d, J = 12.1 Hz, 2H), 3.27-3.22 (m, 2H), 2.82 (s, 3H), 2.49 (s, 3H), 2.44 (t, J = 7.4 Hz, 2H), 2.20 (d, J = 13.5 Hz, 2H), 2.10-2.02 (m, 2H), 1.89 (p, J = 7.0 Hz, 2H), 1.82-1.75 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 171.41, 158.77 (q, J = 31.3 Hz), 157.40, 154.02, 153.37, 149.33, 144.52, 141.97, 139.97, 126.17, 125.74, 124.13, 120.60, 118.62, 116.84, 116.64, 114.65, 110.49, 103.20, 92.53, 69.00, 57.06, 53.01, 48.19, 43.20, 35.76, 28.97, 28.33, 22.31, 14.16. HRMS (ESI): calcd for m / z C 32 H 39 N5O4, [M+H] + 558.3075, found: 558.3065.

[0137] Preparation of compound Y8 of Example 8

[0138] The compounds were prepared according to the procedure of Example 7. The compounds were characterized: 1H NMR (600 MHz, DMSO-d6) δ 9.18 (s, 1H), 7.68 (s, 1H), 7.36 (s, 1H), 7.29 (s, 1H), 7.17 (d, J = 7.8 Hz, 1H), 6.88 (t, J = 7.7 Hz, 1H), 6.84 (s, 1H), 6.71 (d, J = 8.0 Hz, 1H), 6.53 (t, J = 7.5 Hz, 1H), 6.35 (s, 1H), 4.85 (s, 1H), 4.12-4.10 (m, 2H), 4.01-3.96 (m, 2H), 3.94 (s, 3H), 2.72 (s, 3H), 2.42 (s, 3H), 2.37 (t, J = 7.0 Hz, 2H), 2.19-2.12 (m, 2H), 2.01-1.99 (m, 2H), 1.85 (t, J = 7.5 Hz, 2H), 1.69 (t, J = 7.7 Hz, 2H), 1.51 (t, J = 7.8 Hz, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 172.25, 159.10 (q, J = 31.7 Hz),. 155.60, 152.85, 151.22, 148.76, 142.37, 126.57, 126.03, 123.76, 120.40, 118.43, 116.98, 116.54, 116.45, 113.89, 111.27, 109.72, 102.41, 92.89, 68.83, 56.86, 53.06, 47.37, 43.12, 39.98, 36.04, 28.49, 25.55, 25.47, 14.02. HRMS (ESI): calcd for m / z C 33 H 41 N5O4, [M+H] + 572.3231, found: 572.3223.

[0139] Preparation of compound Y9 of Example 9

[0140] The compounds were prepared according to the procedure of Example 7. The compounds were characterized: 1H NMR (600 MHz, DMSO-d6) δ 9.76 (s, 2H), 9.21 (s, 1H), 8.61 (s, 1H), 7.97 (s, 1H), 7.89 (d, J = 3.5 Hz, 1H), 7.78 (s, 1H), 7.18 (dd, J = 7.8, 1.5 Hz, 1H), 7.04 (s, 1H), 6.92-6.88 (m, 1H), 6.75-6.72 (m, 1H), 6.57-6.53 (m, 2H), 4.27 (d, J = 8.1 Hz, 1H), 4.15 (t, J = 6.6 Hz, 2H), 3.99 (s, 3H), 3.57 (d, J = 12.0 Hz, 2H), 3.28-3.20 (m, 3H), 2.80 (s, 3H), 2.51 (s, 3H), 2.37 (t, J = 7.4 Hz, 2H), 2.22-2.18 (m, 4H), 1.87 (q, J = 7.3 Hz, 2H), 1.66 (p, J = 7.4 Hz, 2H), 1.51 (p, J = 7.4 Hz, 2H), 1.44 (q, J = 7.7 Hz, 2H). 13 CNMR (151 MHz, DMSO-d6) δ 171.63, 158.49 (q, J = 31.0 Hz), 157.37, 154.04, 149.35, 142.08, 126.11, 125.72, 124.20, 118.73, 117.17, 116.81, 116.74, 116.49, 114.75, 110.58, 110.52, 103.41, 92.49, 69.23, 57.24, 52.99, 48.28, 43.19, 36.15, 28.87, 28.84, 28.59, 25.72, 25.70, 14.22. HRMS (ESI): calcd for m / z C 34 H 43 N5O4, [M+H] + 586.3388, found: 586.3382.

[0141] Preparation of compound Y10 of Example 10

[0142] The compound was prepared according to the procedure of Example 7. The compound was characterized: 1H NMR (600 MHz, DMSO-d6) δ 9.19 (s, 1H), 7.70 (s, 1H), 7.38 (s, 1H), 7.33 (s, 1H), 7.17 (dd, J = 7.8, 1.5 Hz, 1H), 6.88 (td, J = 7.6, 1.5 Hz, 1H), 6.84 (s, 1H), 6.71 (dd, J = 8.0, 1.4 Hz, 1H), 6.53 (td, J = 7.5, 1.4 Hz, 1H), 6.35 (d, J = 2.8 Hz, 1H), 4.87 (s, 1H), 4.09 (t, J = 6.6 Hz, 2H), 3.99 (s, 1H), 3.94 (s, 3H), 3.44 - 3.40 (m, 3H), 3.11 (s, 1H), 2.72 (s, 3H), 2.42 (s, 3H), 2.34 (t, J = 7.4 Hz, 2H), 2.16 - 2.14 (m, 2H), 2.05 - 2.00 (m, 2H), 1.80 (p, J = 6.8 Hz, 2H), 1.62 (p, J = 7.4 Hz, 2H), 1.46 (q, J = 7.4 Hz, 2H), 1.42 - 1.34 (m, 4H). 13 C NMR (151 MHz, DMSO-d6) δ 171.65, 158.72 (q, J = 32.4 Hz), 154.36, 152.38, 150.40, 148.53, 142.30, 126.06, 125.67, 124.08, 120.69, 118.70, 116.72, 116.57, 116.33, 114.73, 111.70, 109.39, 102.57, 92.87, 68.71, 56.86, 53.04, 47.69, 43.26, 40.51, 36.21, 29.11, 29.01, 28.87, 25.92, 25.76, 14.15. HRMS (ESI): calcd for m / z C 35 H 45 N5O4, [M+H] + 600.3544, found: 600.3537.

[0143] Preparation of compound Y11 of Example 11

[0144] The compounds were prepared according to the procedure of Example 7. The compounds were characterized: 1H NMR(400MHz,DMSO-d6)δ9.20(s,1H),7.89(s,1H),7.66(s,1H),7.30(s,2H),7.17(dd,J=7.9,1.5Hz,1H ),6.92-6.85(m,2H),6.74-6.69(m,2H),6.53(td,J=7.5,1.5Hz,1H),4.89(s,1H),4.11(t,J=6.5Hz,2H) ,4.02-3.89(m,2H),3.93(s,3H),3.45(s,3H),3.17-3.14(m,1H),2.74(s,3H),2.37(t,J=7.3Hz,2H),2. 16-2.19(m,2H),2.05-1.92(m,2H),1.84(p,J=6.9Hz,2H),1.69(p,J=7.4Hz,2H),1.52(q,J=7.9Hz,2H). 13 C NMR(101MHz,DMSO-d6)δ171.59,158.78(q,J=31.1Hz),152.19,150.01,148.60,144.65,142.32,126.09,125.72,124.06,122.18,119.2 0,116.58,116.32,116.22,113.24,112.88,112.09,102.40,93.40,68.59,56.80,47.55,36.19,28.77,25.73,25.56.HRMS(ESI):calcd for m / zC 32 H 40 N5O4, [M+H] + 558.3075, found: 558.3077.

[0145] Example 12

[0146] DNA methyltransferase inhibitory activity assay

[0147] S-adenosylmethionine (S-Adenosylmethionine) was labeled with an isotope. 3 H-SAM was used to test the inhibitory activity of the compound against DNA methyltransferase (DNMT1), with CM-272 as a positive control. The compound and a specific amount of the corresponding DNA methyltransferase isoform were pre-mixed and incubated at room temperature for 15 min. Then, a synthetically produced biotin-labeled oligonucleotide substrate was added. 3H-SAM was reacted at room temperature for 4 h. Then the reaction was transferred to a streptavidin-coated high-throughput plate and reacted at room temperature for 1 h. Radioactivity was measured by liquid scintillation counting. The inhibitory activity of the compound on the enzyme was obtained by comparing with the negative control group (no inhibitor group), and the results are shown in Table 1.

[0148] Histone deacetylase inhibitory activity test

[0149] HDAC1, one of the subtypes of the histone deacetylase family, was used as the research object to test the inhibitory activity of the compound on histone deacetylase. Ten concentration gradients were set for each compound, and three replicates were used. The HDAC inhibitor SAHA was used as a positive control. First, the compound was dissolved in the reaction buffer, then a certain volume of buffer solution containing histone deacetylase was added, and the reaction was incubated at room temperature for 15 min. Then, trypsin and acetylated peptide buffer solution were added as reaction substrates to start the deacetylation reaction, and the compound concentration and enzyme content were set to the desired value. After gentle mixing for 60 s, the reaction was incubated at room temperature, and the kinetic parameters were recorded within 1 h at a certain excitation and emission wavelength. The inhibitory activity of the compound on the enzyme was obtained by comparing with the negative control group (no inhibitor group), and the half-inhibitory concentration (IC 50 ) of the compound on histone deacetylase was calculated, and the results are shown in Table 1.

[0150] Table 1 Enzyme inhibitory activity test of compounds Y1-Y11

[0151]

[0152]

[0153] Example 13

[0154] MTT method for testing cell proliferation inhibitory activity

[0155] The inhibitory effect of the compound on breast cancer cell lines MDA-MB-231 and MDA-MB-468 was tested, and positive compounds Decitabine (DNMT inhibitor), CM-272 (DNMT and G9a dual-target inhibitor) and SAHA (HDAC inhibitor) were used as positive controls. The specific steps of the test include:

[0156] (a) Sample preparation: The compound was prepared as a 10 mM DMSO solution, and the sample solution was diluted to the corresponding concentration with a DMSO gradient

[0157] (b) Plating: Take the logarithmic phase cells, count them by hemocytometer, and inoculate the tumor cells into a 96-well plate at a density of 6-8 x 10 3 cells per well, with a medium volume of 99 μL per well;

[0158] (c) Drug addition: 12h after plating (after cell attachment), 1 μΐ of test compound solution was added to each well to achieve the final concentration of the set value, and 4 replicates were set for each concentration. Meanwhile, three positive drug groups (HDAC inhibitor SAHA or DNMT inhibitor Decitabine, DNMT and G9a dual-target inhibitor CM-272 were added respectively) and one blank group (1 μΐ of DMSO was added) were set;

[0159] (d) MTT treatment: 72h after incubation of the compound with the cells, MTT solution (5 mg / mL, 10 μΐ / well) was added to the experimental and control groups, and the 96-well plate was incubated in the incubator for another 4h;

[0160] (e) Post-treatment and OD value testing: the 96-well plate was centrifuged and the culture medium was removed, DMSO (100 μΐ / well) was added, and the 96-well plate was shaken on a microshaker for 5 min, and then the OD value at 490 nm was tested using an enzyme labeler. Finally, the inhibition rate of the compound on the proliferation of tumor cells at the corresponding concentration was calculated according to the OD value.

[0161] Test results (Table 2)

[0162] Table 2: Test of the inhibitory activity of the compound on the proliferation of tumor cells

[0163]

[0164] From the results of the above DNA methyltransferase inhibition activity test, histone deacetylase inhibition activity test and tumor cell proliferation inhibition activity test, it can be seen that the quinoline derivative of the present application has good inhibition activity on DNA methyltransferase and histone deacetylase, and can effectively inhibit the proliferation of tumor cells, and can be used as a lead compound for the development of anti-tumor drugs.

[0165] The above has described various embodiments of the present application, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and changes are obvious to those skilled in the art without departing from the scope and spirit of the described embodiments.

Claims

1. A quinoline derivative, characterized in that, The quinoline derivative is selected from at least one compound shown in the structure of formula I or a tautomer thereof or a mixture thereof, or a prodrug thereof, or a pharmaceutically acceptable salt, solvate or hydrate thereof; R1 is a substituted or unsubstituted alkyl, alkenyl, alkynyl or aryl group; R2 is a nitrogen-containing substituent; One of M1 and M2 is a group derived from a substituted or unsubstituted six-membered ring, and the other is a chemical bond or a hydroxyl group; preferably, the six-membered ring is a piperidine ring, a pyrimidine ring or a benzene ring, and the substituted group is an amine group; m is an integer from 0 to 5, n is an integer from 0 to 5, and p is 0 or 1.

2. The quinoline derivative according to claim 1, wherein, The compound shown in the structure of formula I is a compound shown in formula 1, formula 2 or formula 3; n1, n2 and n3 are each independently 1, 2, 3 or 4; and R3 is an amine group.

3. The quinoline derivative according to claim 1 or 2, wherein, In the R1 substituent, The alkyl group is a C1-C10 alkyl group, preferably a C1-C5 alkyl group; The alkenyl group is a C2-C10 alkenyl group, preferably a C2-C5 alkenyl group; The alkynyl group is a C2-C10 alkynyl group, preferably a C2-C5 alkynyl group; The aryl group is a phenyl group, a five-membered aromatic ring group, or a six-membered aromatic ring group; the five-membered aromatic ring group is preferably a furan group, a thiophene group, an imidazole group, a pyrrole group or a pyrazole group; and the six-membered aromatic ring group is preferably a pyridine group or a pyrimidine group; The substituted group is selected from a C1-C5 alkyl group, a C1-C5 alkoxy group, a C2-C5 alkenyl group, a C2-C5 alkynyl group, a halogen, a hydroxyl group, a nitro group, a trifluoromethyl group, a trifluoromethoxy group, an amino group, an acrylamide group or a cyano group; R2 is a substituted or unsubstituted pyrrolidine group or a substituted or unsubstituted piperidine group; preferably, it is selected from at least one of the following groups:

4. The quinoline derivative according to claim 1, wherein, The pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt; Preferably, the inorganic acid salt is selected from any one of the following inorganic acids: hydrochloric acid, sulfuric acid and phosphoric acid; Preferably, the organic acid salt is selected from any one of the following organic acids: acetic acid, trifluoroacetic acid, malonic acid, citric acid and p-toluenesulfonic acid.

5. The quinoline derivative according to any one of claims 1 to 4, wherein, The quinoline derivative is selected from at least one of the following compounds:

6. A process for the preparation of a quinoline derivative according to any one of claims 1-5, characterized in that, It comprises the following steps: (1) reacting 5-amino-2-methoxyphenol with excess malonic acid in a solution of phosphorus oxychloride to obtain 2,4-dichloro-6-methoxy-quinolin-7-ol; and / or, (2) introducing an alkylene segment through a nucleophilic substitution reaction; and / or, (3) introducing a group derived from a substituted or unsubstituted six-membered ring through a nucleophilic substitution reaction; and / or, (4) introducing the R1 group through a Suzuki coupling reaction; and / or, (5) introducing the R2 group through a Buchwald-Hartwig coupling reaction.

7. The process for the preparation of quinoline derivatives according to claim 6 wherein, The compound shown in formula 1 sequentially introduces a piperidine ring, an alkylene segment, an R1 group and an R2 group; Preferably, the compound shown in formula 2 sequentially introduces an alkylene segment, an R1 group, an imine-substituted pyrimidine ring and an R2 group; Preferably, the compound shown in formula 3 sequentially introduces an alkylene segment, an R1 group, an R2 group and an amine-substituted benzene ring; Preferably, in step (1), the reaction temperature is 80-150°C, and the reaction time is 20-30 h. Preferably, in step (4), the Suzuki coupling reaction is carried out in a mixed solvent of an organic solvent and water, using a palladium catalyst and at least one of potassium phosphate, sodium carbonate, potassium carbonate or cesium carbonate as a base; the reaction temperature is 50-150°C, and the reaction time is 1-10h; Preferably, in step (5), the Buchwald-Hartwig coupling reaction is carried out in an organic solvent, using a palladium catalyst and at least one of potassium phosphate, sodium carbonate, potassium carbonate or cesium carbonate as a base, the organic solvent is preferably at least one of dimethylformamide, chloroform, dichloromethane, acetone, tetrahydrofuran and acetonitrile; the reaction temperature is 80-200°C, and the reaction time is 20-40h.

8. A pharmaceutical composition comprising the quinoline derivative according to any one of claims 1-5 as an active ingredient and a pharmaceutically acceptable adjuvant. The pharmaceutical composition is preferably in a form selected from the group consisting of an injection, a tablet, a powder, a granule, a capsule, an oral solution, a paste, a cream.

9. Use of the quinoline derivative according to any one of claims 1-5 and / or the pharmaceutical composition according to claim 8 in at least one of the following aspects: 1) preparation of a DNA methyltransferase and histone deacetylase inhibitor; 2) preparation of a eukaryotic tumor cell proliferation inhibitor; 3) preparation of a drug for preventing and / or treating a tumor.

10. The use according to claim 9, wherein the DNA methyltransferase comprises a subtype selected from the group consisting of DNMT1, DNMT3A and DNMT3B in a mammalian cell; the histone deacetylase comprises a subtype selected from the group consisting of HDAC1, HDAC2, HDAC3, HDAC8, HDAC4, HDAC5, HDAC7, HDAC9, HDAC6, HDAC10 and HDAC11 in a mammalian cell; the eukaryote is a mammal; the tumor cell is a cancer cell; the cancer cell is preferably selected from the group consisting of a leukemia cancer cell, a breast cancer cell, a liver cancer cell, a pancreatic cancer cell, a lung cancer cell, a brain cancer cell, an ovarian cancer cell, an uterine cancer cell, a testicular cancer cell, a skin cancer cell, a stomach cancer cell, a nasopharyngeal cancer cell, a colon cancer cell, a bladder cancer cell or a rectal cancer cell; more preferably, the cancer cell is a human chronic myelogenous leukemia cell and / or a human histiocytic lymphoma cell; the leukemia cancer cell is preferably a human chronic myelogenous leukemia cell line K562, the lymphoma cell is preferably a human histiocytic lymphoma cell U937, the lung cancer cell is preferably a human lung cancer cell NCI-H520, the human brain glioma cell is preferably U251, the melanoma cancer cell is preferably A375, the glioblastoma cell is preferably a human glioblastoma cell A172 and a human brain astrocytic glioblastoma cell U-118MG, the cervical cancer cell is preferably a human cervical cancer cell line Hela, the nasopharyngeal cancer cell is preferably a nasopharyngeal cancer cell line CNE-2, the liver cancer cell is preferably a human liver cancer cell line HepG2, the breast cancer cell is preferably a human breast cancer cell MCF-7 and MDA-MB-231; The tumor is a carcinoma; the carcinoma is a leukemia, lymphoma, lung cancer, melanoma, glioblastoma, cervical cancer, nasopharyngeal cancer, liver cancer, breast cancer, brain cancer, pancreatic cancer, ovarian cancer, uterine cancer, testicular cancer, skin cancer, stomach cancer, colon cancer, bladder cancer, or rectal cancer. The tumor is a carcinoma; the carcinoma is a leukemia, lymphoma, lung cancer, melanoma, glioblastoma, cervical cancer, nasopharyngeal cancer, liver cancer, breast cancer, brain cancer, pancreatic cancer, ovarian cancer, uterine cancer, testicular cancer, skin cancer, stomach cancer, colon cancer, bladder cancer, or rectal cancer. The tumor is a carcinoma; the carcinoma is a leukemia, lymphoma, lung cancer, melanoma, glioblastoma, cervical cancer, nasopharyngeal cancer, liver cancer, breast cancer, brain cancer, pancreatic cancer, ovarian cancer, uterine cancer, testicular cancer, skin cancer, stomach cancer, colon cancer, bladder cancer, or rectal cancer. The tumor is a carcinoma; the carcinoma is a leukemia, lymphoma, lung cancer, melanoma,

Images