Dihydropyridine derivative as well as preparation method, intermediate, composition and application thereof

Abstract

The invention discloses a dihydropyridine derivative as well as a preparation method, an intermediate, a composition and application thereof. The structure of the dihydropyridine derivative is shown as a formula (I), wherein in the formula # imgabs0, R1 is selected from any one of-H,-CH3,-CH2CH3 and-OCH3; r < 2 > is selected from any one of-H,-CH3 and-CH2CH3; r3 is selected from any one of-H,-OH,-Br and-NH2; r4 is selected from any one of-H,-CH3,-Br,-Cl,-CF3,-OH and-NH2; r < 5 > is selected from any one of-H,-CH3,-Br,-Cl,-CF3,-OCH3 and-NH2; r6 is selected from any one of-H,-CH3,-Br,-F,-CF3 and-NH2; and R7 is selected from any one of-H,-CH3,-NO2 and-NH2. The dihydropyridine derivative has the advantages of excellent calcium channel retardation activity, remarkable P-glycoprotein inhibition effect, high bioavailability, simple and convenient synthesis process and the like, can be used as a novel antihypertensive-anticancer synergistic treatment drug, and efficiently realizes dual intervention of blood pressure regulation and tumor chemotherapy sensitization.

Description

Technical Field

[0001] The present application relates to the technical field of chemical synthesis, and in particular to a dihydropyridine derivative and a preparation method, intermediate, composition and application thereof. Background Art

[0002] The prevalence of hypertension and cancer remains high among the elderly, and co-morbidity (comorbidity) of hypertension and cancer is common in clinical practice. The coexistence of these two conditions significantly increases health risks, and their interaction may exacerbate disease progression. During cancer treatment, patients with severe hypertension may be unable to receive certain chemotherapy or immunotherapy treatments, while patients undergoing surgery are more prone to hemodynamic fluctuations.

[0003] In addition, for patients with both hypertension and cancer, clinical treatment options typically involve separate drug interventions for the two diseases. However, this treatment strategy requires patients to take multiple prescription drugs over a long period of time, which not only increases the risk of complex side effects but can also significantly weaken the effectiveness of chemotherapy due to drug resistance.

[0004] Dihydropyridines are a class of calcium channel blockers (CCBs) that selectively target vascular smooth muscle. By blocking L-type calcium channels and reducing calcium influx, they dilate blood vessels and lower blood pressure. They are widely used to treat cardiovascular diseases such as hypertension and angina pectoris. However, existing dihydropyridines are primarily targeted at treating patients with hypertension and cannot simultaneously address the needs of treating both hypertension and enhancing tumor chemotherapy sensitization.

[0005] Therefore, how to achieve efficient synchronous treatment for patients with hypertension and cancer has become an important issue that needs to be urgently addressed in the fields of clinical medicine and scientific research. Summary of the Invention

[0006] In view of this, the technical problem to be solved in this application is to address the shortcomings of existing dihydropyridine drugs in the art, which have a single function and cannot simultaneously meet the needs of hypertension treatment and tumor chemotherapy sensitization. The present application provides a new multifunctional dihydropyridine calcium channel / P-glycoprotein dual-target modulator, namely a new dihydropyridine derivative. This dihydropyridine derivative has the advantages of excellent calcium channel blocking activity, significant P-glycoprotein inhibition effect, high bioavailability, and simple synthesis process. It can be used as a new antihypertensive and anticancer synergistic therapeutic drug, effectively achieving dual intervention of blood pressure regulation and tumor chemotherapy sensitization.

[0007] To achieve the above objectives, this application provides the following technical solutions:

[0008] In a first aspect, the present application provides a dihydropyridine derivative, a pharmaceutically acceptable salt or stereoisomer thereof, wherein the structure of the dihydropyridine derivative is shown in the following formula (I):

[0009]

[0010] Wherein, R1 is selected from any one of -H, -CH3, -CH2CH3, and -OCH3;

[0011] R2 is selected from any one of -H, -CH3, -CH2CH3;

[0012] R3 is selected from any one of -H, -OH, -Br, and -NH2;

[0013] R4 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OH, -NH2;

[0014] R5 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OCH3, -NH2;

[0015] R6 is selected from any one of -H, -CH3, -Br, -F, -CF3, -NH2;

[0016] R7 is selected from any one of -H, -CH3, -NO2, and -NH2.

[0017] The compound represented by formula (I) introduces an aromatic amine substituent onto the dihydropyridine core, creating a "dihydropyridine-arylamine" hybrid structure, forming a novel dihydropyridine derivative. While retaining the inherent calcium channel blocking activity of the dihydropyridine core, this dihydropyridine derivative achieves dual inhibitory effects on L-type calcium channels and P-glycoprotein by introducing an aromatic amine side chain.

[0018] In designing the molecular structure of the compound represented by formula (I), this application focused on optimizing the electron cloud distribution of the dihydropyridine core, the spatial configuration of the aromatic amine side chain, and the flexible length of the linker group, thereby ensuring that the compound can simultaneously and efficiently bind to the two target proteins mentioned above, achieving synergistic pharmacological effects of regulating calcium influx into vascular smooth muscle and inhibiting drug efflux from tumor cells. This innovative dual-target design strategy provides a new treatment option for patients with hypertension and cancer.

[0019] The compounds shown in formula (I) designed in this application, and their pharmaceutically acceptable salts or stereoisomers, can exhibit significant dual pharmacological activity: on the one hand, they effectively lower blood pressure by inhibiting vascular smooth muscle calcium channels, and on the other hand, they significantly enhance the sensitivity of tumor cells to chemotherapy drugs by blocking the function of P-glycoprotein. It is particularly noteworthy that these molecules can not only directly inhibit P-glycoprotein-mediated drug efflux, but also enhance chemotherapy-induced cell apoptosis by regulating intracellular calcium signaling pathways, thereby achieving multiple synergistic therapeutic effects. This innovative dual-target drug design strategy breaks through the limitations of traditional single-target drugs and provides a safer and more effective treatment option for patients with hypertension and cancer.

[0020] In certain preferred embodiments, the structure of the aforementioned dihydropyridine derivative is shown in the following formula (II):

[0021]

[0022] Wherein, R1 is selected from any one of -H, -CH3, -CH2CH3, and -OCH3;

[0023] R5 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OCH3, and -NH2.

[0024] In the dihydropyridine derivative represented by the above formula (II), i.e., HAC-1, i.e., the compound represented by formula (I), R1 is selected from any one of -H, -CH3, -CH2CH3, and -OCH3, R2 is -CH3, R3 is -Br, R4 is -H, R5 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OCH3, and -NH2, R6 is -H, and R7 is -NO2.

[0025] In certain preferred embodiments, the structure of the aforementioned dihydropyridine derivative is shown in the following formula (III):

[0026]

[0027] Wherein, R2 is selected from any one of -H, -CH3, -CH2CH3;

[0028] R4 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OH, and -NH2.

[0029] In the dihydropyridine derivative represented by the above formula (III), i.e., HAC-2, i.e., the compound represented by formula (I), R1 is -H, R2 is selected from any one of -H, -CH3, and -CH2CH3, R3 is -H, R4 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OH, and -NH2, R5 is -OCH3, R6 is -H, and R7 is -NH2.

[0030] In certain preferred embodiments, the structure of the aforementioned dihydropyridine derivative is shown in the following formula (IV):

[0031]

[0032] Wherein, R1 is any one of -H, -CH3, and -OCH3;

[0033] R6 is selected from any one of -H, -CH3, -Br, -F, -CF3, and -NH2.

[0034] The dihydropyridine derivative represented by the above formula (IV), i.e., HAC-3, is a compound represented by formula (I), wherein R1 is any one of -H, -CH3, and -OCH3, R2 is -CH3, R3 is -H, R4 is -CF3, R5 is -H, R6 is selected from any one of -H, -CH3, -Br, -F, -CF3, and -NH2, and R7 is -NO2.

[0035] In a second aspect, the present application provides a method for preparing a dihydropyridine derivative, which includes at least one of methods 1 to 3, wherein:

[0036] Method 1, for preparing the aforementioned HAC-1 compound, comprises the following steps:

[0037]

[0038] Wherein, R1 is any one of -H, -CH3, and -OCH3;

[0039] R5 is any one of -H, -CH3, -Br, -Cl, -CF3, -OCH3, -NH2;

[0040] 4-Hydroxy-3-nitrobenzaldehyde is first reacted with ammonium acetate and β-keto acid to obtain intermediate 1, which is then dissolved in aqueous sodium hydroxide solution, followed by dropwise addition of chloroacetic acid and saturated sodium hydroxide solution to obtain intermediate 2. Intermediate 2 is then dissolved in methanol, followed by addition of aqueous sodium hydroxide solution to obtain intermediate 3. The prepared intermediate 3 and the corresponding aniline compound are then dissolved in dichloromethane, followed by addition of N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine to obtain HCA-1 compound.

[0041] Method 2, for preparing the aforementioned HAC-2 compound, comprises the following steps:

[0042]

[0043] Wherein, R2 is selected from any one of -H, -CH3, -CH2CH3;

[0044] R4 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OH, -NH2;

[0045] 9-Fluorenone and 3-hydroxy-4-aminobenzylamine are first reacted with β-ketoester to prepare intermediate 4, which is then dissolved in water with sodium hydroxide and reacted with chloroacetic acid to prepare intermediate 5. Intermediate 5 and the corresponding amine are dissolved in dichloromethane, and N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine are added to react to obtain HCA-2 compound.

[0046] Method 3, for preparing the aforementioned HAC-3 compound, comprises the following steps:

[0047]

[0048] Wherein, R1 is selected from any one of -H, -CH3, -CH2CH3, and -OCH3;

[0049] R6 is selected from any one of -H, -CH3, -Br, -F, -CF3, -NH2;

[0050] 4-Hydroxy-3-nitrobenzaldehyde is first reacted with β-ketoester and ammonium acetate to prepare intermediate 1, which is then dissolved in N,N-dimethylformamide with bromopropyl alcohol and potassium carbonate in appropriate proportions to prepare intermediate 6. Intermediate 6, 2-iodoacylbenzoic acid and iodine are then dissolved in N,N-dimethylformamide to react to prepare intermediate 7. Intermediate 7 and the corresponding aniline derivative are then dissolved in dichloromethane, and N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine are added to react to obtain HCA-3 compound.

[0051] In certain preferred embodiments, the specific steps of method 1 are as follows:

[0052] 1) 4-Hydroxy-3-nitrobenzaldehyde, β-keto acid, and ammonium acetate are mixed and dissolved in ethanol in appropriate proportions and refluxed for 16-18 hours. After the reaction is complete, the mixture is cooled, filtered, and recrystallized from ethanol. The solid is filtered and dried to obtain intermediate 1; wherein the molar ratio of 4-hydroxy-3-nitrobenzaldehyde, β-keto acid, and ammonium acetate is 1-1.5:2-4:2-4;

[0053] 2) Dissolve Intermediate 1 and ethyl chloroacetate in acetonitrile, add potassium iodide and anhydrous potassium carbonate, and react at 100°C for 8 hours. After the reaction is complete, cool to room temperature. Filter, wash the solid with ethyl acetate, extract the filtrate with brine and ethyl acetate, dry the organic layer over anhydrous sodium sulfate, concentrate, and purify by column chromatography to obtain Intermediate 2; the molar ratio of Intermediate 1, ethyl chloroacetate, potassium iodide, and anhydrous potassium carbonate is 1:1-2:0.5:3.

[0054] 3) Dissolve intermediate 2 in methanol, add 6 mol / L aqueous sodium hydroxide solution, and reflux for 8 hours. After the reaction is complete, concentrate the solvent under reduced pressure, adjust the pH to 2 with 6 mol / L aqueous hydrochloric acid, extract with dichloromethane, dry over anhydrous sodium sulfate, concentrate, and recrystallize from ethanol to obtain intermediate 3.

[0055] 4) Intermediate 3 and its corresponding aniline derivative were dissolved in anhydrous dichloromethane, and N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine were added. The reaction was allowed to proceed at room temperature for 18 hours. After the reaction was complete, the mixture was filtered, and the filtrate was extracted with dichloromethane. The organic layer was washed sequentially with dilute hydrochloric acid, saturated aqueous sodium bicarbonate solution, and water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain HCA-1. The molar ratio of intermediate 3, the corresponding aniline derivative, N,N-dicyclohexylcarbodiimide, and 4-dimethylaminopyridine was 1:1-1.5:1-2:0.2.

[0056] In certain preferred embodiments, the specific steps of the aforementioned method 2 are as follows:

[0057] 1) Under N2 conditions, 3-hydroxy-4-aminobenzylamine was added to a toluene solution of 9-fluorenone, and the mixture was stirred at room temperature for 1 hour. β-keto acid and ammonium acetate were then added, and the mixture was refluxed for 20 hours. After the reaction was complete, the mixture was evaporated under reduced pressure and purified by column chromatography to obtain intermediate 4; wherein the molar ratio of 9-fluorenone, 3-hydroxy-4-aminobenzylamine, β-keto acid, and ammonium acetate was 0.5:1:1-2:1-2;

[0058] 2) Dissolve intermediate 4 and sodium hydroxide in water, and add saturated chloroacetic acid solution and saturated sodium hydroxide solution dropwise at room temperature, controlling the addition rate to maintain the pH at 9-10. Heat to 80-90°C and react for 3 hours. After the reaction is complete, adjust the pH of the mixture to 1 with hydrochloric acid, filter the mixture, wash with water, dry, and then recrystallize from petroleum ether to obtain intermediate 5; wherein the molar ratio of intermediate 4, sodium hydroxide, and chloroacetic acid is 1:1:1.5;

[0059] 3) Intermediate 5 and its corresponding aniline derivative were dissolved in anhydrous dichloromethane, and N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine were added. The reaction was allowed to proceed at room temperature for 18 hours. After the reaction was complete, the mixture was filtered, and the filtrate was extracted with dichloromethane. The organic layer was washed sequentially with dilute hydrochloric acid, saturated aqueous sodium bicarbonate, and water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain HCA-2. The molar ratio of intermediate 5, the corresponding aniline derivative, N,N-dicyclohexylcarbodiimide, and 4-dimethylaminopyridine was 1:1-1.5:1-2:0.2.

[0060] In certain preferred embodiments, the specific steps of the aforementioned method 3 are as follows:

[0061] 1) 4-Hydroxy-3-nitrobenzaldehyde, β-ketoester, and ammonium acetate are mixed in appropriate proportions and dissolved in ethanol. The mixture is refluxed for 16-18 hours. After the reaction is complete, the mixture is cooled, filtered, and recrystallized from ethanol. The solid is filtered and dried to obtain Intermediate 1. The molar ratio of 4-hydroxy-3-nitrobenzaldehyde, β-ketoester, and ammonium acetate is 1-1.5:2-4:2-4.

[0062] 2) Dissolve intermediate 1, bromopropanol, and potassium carbonate in N,N-dimethylformamide and react at 80-100°C for 8 hours. After the reaction is complete, cool to room temperature. After filtering, adjust the pH of the mixture to neutral with hydrochloric acid, extract with ethyl acetate, and dry the organic layer over anhydrous sodium sulfate. Concentrate and purify by column chromatography to obtain intermediate 6; the molar ratio of intermediate 1 to bromopropanol and potassium carbonate is 1:1-2:1-2.

[0063] 3) Dissolving intermediate 6, 2-iodoacetylbenzoic acid, and iodine in N,N-dimethylformamide and heating at 100°C for 18 hours. After the reaction is complete, the reaction mixture is cooled to room temperature, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and separated by column chromatography to obtain intermediate 7; wherein the molar ratio of intermediate 6, 2-iodoacetylbenzoic acid, and iodine is 0.25:2:0.4;

[0064] 4) Intermediate 7 and its corresponding aniline derivative were dissolved in anhydrous dichloromethane, and N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine were added. The reaction was allowed to proceed at room temperature for 18 hours. After the reaction was complete, the mixture was filtered and the filtrate was extracted with dichloromethane. The organic layer was washed sequentially with dilute hydrochloric acid, saturated aqueous sodium bicarbonate solution, and water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain HCA-3. The molar ratio of intermediate 7, the corresponding aniline derivative, N,N-dicyclohexylcarbodiimide, and 4-dimethylaminopyridine was 1:1-1.5:1-2:0.2.

[0065] In a third aspect, the present application provides an intermediate for preparing the aforementioned dihydropyridine derivative, wherein the intermediate is at least one of the following intermediates 1-7;

[0066]

[0067] Wherein, R1 is selected from any one of -H, -CH3, -CH2CH3, and -OCH3;

[0068] R2 is selected from any one of -H, -CH3, and -CH2CH3.

[0069] Among the above 7 intermediates, intermediate 1, intermediate 2, and intermediate 3 can be used to prepare HAC-1 compound; intermediate 4 and intermediate 5 can be used to prepare HAC-2 compound; intermediate 1, intermediate 6, and intermediate 7 can be used to prepare HAC-3 compound.

[0070] In a fourth aspect, the present application provides a pharmaceutical composition comprising the aforementioned dihydropyridine derivative, a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier, excipient or diluent.

[0071] The dihydropyridine derivatives include mixtures of the aforementioned specific dihydropyridine derivatives in any proportion.

[0072] In a fifth aspect, the present application provides the use of the aforementioned dihydropyridine derivatives, pharmaceutically acceptable salts or stereoisomers thereof, or pharmaceutical compositions thereof, in the preparation of drugs for treating or preventing hypertension combined with cancer.

[0073] Dihydropyridine derivatives, pharmaceutically acceptable salts or stereoisomers thereof, or pharmaceutical compositions thereof can be used as dual-target modulators to achieve dual therapeutic effects of lowering blood pressure and enhancing chemotherapy sensitivity by simultaneously inhibiting L-type calcium channels and blocking P-glycoprotein. The synergistic mechanism of multifunctional dihydropyridine calcium channel / P-glycoprotein dual-target modulators is as follows: Figure 1 This molecule, through its unique "dihydropyridine-arylamine" hybrid structure, selectively binds to the L-type calcium channel of vascular smooth muscle cells through the dihydropyridine core, inhibiting calcium ion influx and achieving a blood pressure-lowering effect. It also targets tumor cell P-glycoprotein through the aromatic amine side chain, blocking its drug efflux function and enhancing the intracellular accumulation and efficacy of chemotherapy drugs.

[0074] The dihydropyridine core in the dual-target modulator is the calcium channel blocking active center, which binds to the calcium channel α1 subunit through its specific spatial conformation; the aromatic amine substituent is the P-glycoprotein inhibitory functional domain, which occupies the P-glycoprotein drug binding pocket through hydrophobic interactions and π-π stacking. The implementation scheme of this application achieves precise regulation of the dual inhibitory activity of calcium channels and P-glycoprotein by systematically optimizing the type and position of the aromatic amine substituents. The dihydropyridine skeleton in the molecule is the structural basis for maintaining the calcium channel blocking activity, while the functionalized aromatic amine side chain is the key to enhancing the inhibitory efficacy of P-glycoprotein. This dual-target design enables a single molecule to act on the cardiovascular system and tumor cells simultaneously, achieving a synergistic therapeutic effect of blood pressure control and chemotherapy sensitization.

[0075] Compared with the prior art, this application has at least the following beneficial effects:

[0076] 1. This application provides a series of novel dihydropyridine derivatives, which achieve the dual functions of calcium channel blocking and P-glycoprotein inhibition through molecular structural innovation, solving the technical problem of traditional treatment requiring combined drug use.

[0077] 2 The dual-target modulator developed in this application can simultaneously intervene in the two major pathological links of hypertension and tumor resistance through a single molecule, providing an innovative solution for the treatment of patients with hypertension and cancer, and has broad application prospects in the field of cardiovascular-tumor synergistic treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0078] Figure 1 This is the intracellular mechanism of action of electron-deficient conjugated dihydropyridine drugs in this application.

[0079] Figure 2 The H NMR spectrum and C NMR spectrum of the HCA-1-1 compound prepared in this example; (a) is the H NMR spectrum, and (b) is the C NMR spectrum.

[0080] Figure 3 The UV absorption spectra of the dihydropyridine derivatives synthesized in the examples of this application at different concentrations.

[0081] Figure 4 This is a molecular docking diagram of the dihydropyridine derivative synthesized in the examples of this application and P-glycoprotein.

[0082] Figure 5 This is a molecular docking diagram of the dihydropyridine derivative synthesized in the examples of this application and the L-type calcium channel.

[0083] Figure 6 This is a graph showing the anticancer properties of the dihydropyridine derivatives synthesized in the examples of this application. DETAILED DESCRIPTION

[0084] In order to make the purpose, technical solutions and advantages of this application more clear, the following further describes this application in detail with reference to the embodiments. It should be understood that the specific embodiments described herein are only used to explain this application and are not intended to limit this application.

[0085] Those skilled in the art will understand that, unless otherwise stated, the terms "said," "the," "the aforementioned," and "above" used in this application may include plural forms. It should be further understood that the term "comprising" used in the specification of this application refers to the presence of the stated features, steps, or operations, but does not exclude the presence or addition of one or more other features, steps, or operations.

[0086] The "room temperature" in this application text may fluctuate in different seasons and is usually between 15 and 30°C.

[0087] Those skilled in the art will understand that if no specific experimental steps or conditions are specified in the examples, the experiments can be carried out according to the conventional experimental steps or conditions described in the literature in the field; if the raw materials or instruments and equipment used do not specify the manufacturer, they are all conventional products that can be purchased commercially.

[0088] Those skilled in the art will understand that, unless otherwise specified herein, when numerical ranges are given in the examples, both endpoints of each numerical range and any value between the two endpoints may be used. Unless otherwise defined, all technical and scientific terms used in this application are consistent with the prior art as understood by those skilled in the art and the description of this application. Any prior art methods, equipment, and materials similar or equivalent to those described in the examples of this application may also be used to implement this application.

[0089] As used herein, the term "pharmaceutically acceptable salt" refers to relatively non-toxic acid addition salts or base addition salts of the compounds of the present invention.

[0090] As used herein, the term "stereoisomer" refers to a stereoisomer of a dihydropyridine derivative in which an asymmetric or chiral center is present. Stereoisomers are designated as (R) or (S) depending on the configuration of substituents around the chiral carbon atom. The embodiments described herein specifically include various stereoisomers and mixtures thereof. Stereoisomers include racemates, enantiomers, diastereomers, and mixtures of enantiomers or diastereomers. In some embodiments, each stereoisomer of a compound is synthesized from commercial raw materials containing an asymmetric or chiral center, or by preparing a racemic mixture and then resolving it. Resolving methods include, for example: (1) combining the mixture of enantiomers with a chiral auxiliary, separating the diastereomeric mixture by recrystallization or chromatography, and releasing the optically pure product from the auxiliary; or (2) directly separating the mixture of optical enantiomers on a chiral chromatographic column.

[0091] In the present application, the dihydropyridine derivative represented by formula (I), its pharmaceutically acceptable salt or stereoisomer can be administered to the subject by any convenient route of administration, which can be systemic / peripheral or at the desired site of action, including but not limited to oral administration (e.g., ingestion), topical administration (including, for example, transdermal, intranasal, ocular, oral and sublingual), pulmonary administration (e.g., inhalation or insufflation of an aerosol through the mouth or nose), rectal administration, vaginal administration, parenteral administration (e.g., injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal administration, intratracheal administration, subcutaneous administration, intraarticular administration, subarachnoid administration and intrasternal administration), and implantation of a depot (e.g., subcutaneous or intramuscular implantation).

[0092] In the present application, the subject of treatment of the dihydropyridine derivative, its pharmaceutically acceptable salt or stereoisomer or pharmaceutical composition can be an animal, a vertebrate, a mammal, a rodent (e.g., a guinea pig, a hamster, a rat, a mouse), a murine (e.g., a mouse), a canine (e.g., a dog), a primate, anthropoid (e.g., a monkey or an ape), a monkey (e.g., a marmoset, a baboon), an ape (e.g., a gorilla, a chimpanzee, an orangutan, a gibbon) or a human.

[0093] In the present application, the pharmaceutical composition comprises the dihydropyridine derivative represented by formula (I) herein, a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier, excipient, or diluent, etc. The dihydropyridine derivative represented by formula (I), a pharmaceutically acceptable salt or stereoisomer thereof, can be mixed with a pharmaceutically acceptable carrier, excipient, diluent, etc. according to standard pharmaceutical procedures to prepare a corresponding pharmaceutical composition, which can be administered to mammals, including humans.

[0094] In the present application, the carrier, excipient and diluent refer to inactive ingredients in the pharmaceutical composition that do not cause significant irritation to the organism and do not interfere with the biological activity of the administered dihydropyridine derivative, specifically including but not limited to water, lactose, glucose, fructose, sucrose, sorbitol, mannitol, polyethylene glycol, propylene glycol, starch, rubber, gelatin, alginate, calcium silicate, calcium phosphate, cellulose, aqueous syrup, methylcellulose, polyvinylpyrrolidone, alkyl parahydroxybenzosorbate, talc, magnesium stearate, stearic acid, glycerin, sesame oil, olive oil, soybean oil and other oils and mixtures thereof.

[0095] In the present application, the dihydropyridine derivatives shown in formula (I), their pharmaceutically acceptable salts or stereoisomers, or pharmaceutical compositions thereof can be prepared into formulations for administration to mammals, including humans. The formulations can be conveniently provided in unit dosage form and are preferably prepared by methods well known in the pharmaceutical field. Such methods include the step of combining the compound of the present application with a carrier constituting one or more auxiliary ingredients. Typically, the dihydropyridine compound shown in formula (I) of the present application is closely combined with a liquid carrier or a finely divided solid carrier or two carriers to prepare the formulation, and then the product is shaped as needed. The formulation can be in the form of a liquid, solution, suspension, emulsion, elixir, syrup, tablet, lozenge, granule, powder, capsule, cachet, pill, ampoule, suppository, vaginal suppository, ointment, gel, paste, cream, spray, aerosol, foam, lotion, oil, bolus, confectionery or aerosol.

[0096] In the application, the cancer includes but is not limited to breast cancer, ovarian cancer, endometrial cancer, cervical cancer, lung cancer, prostate cancer, pancreatic cancer, blood cancer, stomach cancer, gallbladder cancer, liver cancer, head and neck cancer, esophageal cancer, kidney cancer, brain cancer, leukemia, colon cancer, intestinal tumor, glioblastoma, lymphoma or melanoma.

[0097] The technical solution of this application and the technical effects achieved are described in detail below through more specific embodiments.

[0098] Example 1

[0099] This embodiment provides a novel method for preparing a dihydropyridine derivative, and its synthetic route is as follows:

[0100] The specific steps are as follows:

[0101] 1) To a 250 mL round-bottom flask, 4-hydroxy-3-nitrobenzaldehyde (15 mmol, 2.5 g), ethyl acetoacetate (21 mmol, 2.73 g), and ammonium acetate (21 mmol, 1.6 g) were dissolved in 160 mL of ethanol and refluxed for 18 h. After the reaction was complete, the mixture was cooled, filtered, and recrystallized from ethanol. The solid was filtered and dried to obtain Intermediate 1-1.

[0102] 2) To a 250 mL round-bottom flask, add intermediate 1-1 (10.9 mmol, 4.2 g) and ethyl chloroacetate (21.8 mmol, 2.67 g), then add 150 mL of acetonitrile to completely dissolve, add potassium iodide (5.4 mmol, 896 mg) and anhydrous potassium carbonate (32 mmol, 4.41 g), and heat to 100°C for 8 h. After the reaction is complete, cool to room temperature. Filter, wash the solid with ethyl acetate, extract the filtrate with brine and ethyl acetate, dry the organic layer over anhydrous sodium sulfate, concentrate, and purify by column chromatography to obtain intermediate 2-1;

[0103] 3) To a 100 mL round-bottom flask, add intermediate 2-1 (10.9 mmol, 5.5 g), then add 50 mL of methanol to dissolve it completely. Then add 10 mL of 6 mol / L aqueous sodium hydroxide solution and reflux for 8 h. After the reaction is complete, concentrate the solvent under reduced pressure, adjust the pH to 2 with 6 mol / L aqueous hydrochloric acid, extract with dichloromethane, dry over anhydrous sodium sulfate, concentrate, and recrystallize from ethanol to obtain intermediate 3-1.

[0104] 4) To a 100 mL round-bottom flask, intermediate 3-1 (5.34 mmol, 2.5 g) and o-bromoaniline (9.6 mmol, 1.65 g) were added, followed by complete dissolution in 60 mL of anhydrous dichloromethane. N,N-dicyclohexylcarbodiimide (6.4 mmol, 1.32 g) and 4-dimethylaminopyridine (0.267 mmol, 32 mg) were added and allowed to react at room temperature for 18 h. After the reaction was complete, the mixture was filtered and the filtrate was extracted with dichloromethane. The organic layer was washed sequentially with dilute hydrochloric acid, saturated aqueous sodium bicarbonate solution, and water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain HCA-1-1.

[0105] The HCA-1-1 compound prepared in Example 1 is a specific form of the HCA-1 compound, that is, in the HCA-1 compound, R1 is -H and R5 is -H.

[0106] Figure 2 The H NMR spectrum and C NMR spectrum of the dihydropyridine derivative (HCA-1-1 compound) prepared in Example 1; (a) is the H NMR spectrum, and (b) is the C NMR spectrum.

[0107] The characterization data of HCA-1-1 compound by hydrogen nuclear magnetic resonance spectrum are as follows: 1H NMR (400 MHz, DMSO-d6) δ9.40 (s, 1H), 8.93 (s, 1H), 7.84 (dd, J = 8.1, 1.6 Hz, 1H), 7.71-7.61 (m, 2H), 7.47-7.35 (m, 2H), 7.35-7.22 (m, 1H), 7.15 (td, J = 7.7, 1.6 Hz, 1H), 5.25 (s, 1H), 4.94 (s, 2H), 4.86 (s, 1H), 3.99 (qq, J = 10.9, 7.1 Hz, 4H), 2.27 (s, 6H), 1.14 (t, J = 7.1 Hz, 6H).

[0108] The carbon NMR spectrum characterization data of HCA-1-1 compound are as follows: 13C NMR (101 MHz, DMSO-d6) δ 167.06, 166.34, 149.30, 146.53, 142.41, 138.94, 135.62, 134.07, 133.16, 128.69, 127.42, 125.63, 124.34, 116.67, 115.92, 101.68, 68.35, 59.65, 38.88, 18.69, 14.57. This confirms that it is the target product HCA-1-1.

[0109] The dihydropyridine derivative HCA-1-1 prepared in Example 1 was tested for its UV-visible absorption spectrum, docking with calcium channels, docking with P-glycoprotein and analysis, and anticancer properties.

[0110] 1. UV-visible absorption spectrum:

[0111] Using MeOH as the solvent, a 5 mM stock solution of the dihydropyridine derivative HCA-1-1 prepared in Example 1 was prepared and diluted sequentially to 3.3 μM, 5.0 μM, 6.7 μM, 8.3 μM, 10.0 μM, 11.7 μM, 13.3 μM, 15.0 μM, 16.7 μM, and 18.3 μM solutions. After vortex mixing, the absorbance of the target solution was measured in the wavelength range of 190-300 nm to obtain the characteristic UV-visible absorption peak.

[0112] The test results are as follows Figure 3 As shown, solutions of dihydropyridine derivatives with different concentrations have obvious absorption peaks at 203nm and 237nm, and the absorbance increases linearly with increasing concentration (in accordance with the Lambert-Beer law), which is consistent with the photophysical characteristics of the conjugated structure of dihydropyridine molecules, proving that the synthesis of the target dihydropyridine compounds and the preparation of concentration gradient solutions are successful.

[0113] 2. Docking with P-glycoprotein:

[0114] First, the P-glycoprotein crystal structure was obtained from the Protein Data Bank (PDB: 3G60). After removing water molecules and cocrystal ligands using AutoDock Tools software, hydrogenation and charge calculation were performed. After the compound structure was optimized, hydrogenation, charge calculation, and flexible bond setting were performed.

[0115] The docking results of dihydropyridine derivatives with P-glycoprotein showed that the interaction between them and P-glycoprotein at the binding site was as follows: Figure 4 As shown, the minimum binding energy is -8.03 kcal / mol. The drug molecule is embedded in the cavity of P-glycoprotein and connects to the TYR-303 and TYR-949 amino acid residues on P-glycoprotein through hydrophobic interactions. Its binding site is x = 19.3, y = 52.68, and z = -0.1. This demonstrates that the dihydropyridine derivative prepared in Example 1 can effectively target the drug binding pocket of P-glycoprotein and form a stable complex with P-glycoprotein, providing a structural basis for the development of novel P-glycoprotein inhibitors. Molecular docking studies provide important insights into the mechanism of action of these compounds and subsequent structural optimization.

[0116] 3. Docking with calcium channels:

[0117] The crystal structure of the L-type calcium channel Cav1.2 was first obtained from the Protein Data Bank (PDB: 8HMB). After removing water molecules and cocrystal ligands using AutoDock Tools software, hydrogenation and charge calculations were performed. After optimizing the compound structure, hydrogenation, charge calculations, and flexible bond configuration were performed.

[0118] The docking results of dihydropyridine derivatives with L-type calcium channel Cav1.2 show that their interaction with L-type calcium channel Cav1.2 at the binding site is as follows Figure 5 As shown, the minimum binding energy is -7.91 Kcal / mol. The drug molecule is embedded in the cavity of the L-type calcium channel Cav1.2 and connects to the SER-1132 amino acid residue on the L-type calcium channel Cav1.2 through hydrophobic interaction. Its binding site is x = 167.03, y = 164.69, and z = 154.11. This shows that the dihydropyridine derivative molecule prepared in Example 1 can effectively target the drug binding site of the L-type calcium channel and can form a stable complex with the L-type calcium channel, providing a structural basis for the development of new L-type calcium channel inhibitors. Molecular docking studies provide an important basis for understanding the mechanism of action of these compounds and subsequent structural optimization.

[0119] 4. Anti-cancer properties:

[0120] To test the anticancer activity of the dihydropyridine derivative HCA-1-1 prepared in Example 1, cisplatin was used as a control group and cisplatin + compound HCA-1 combination was used as an experimental group to test its inhibitory activity against different cancer cell lines (4T1, MDA-MB-231, HeLa, HepG2, SW480).

[0121] The test results are as follows Figure 6 As shown in the figure, it can be seen that HCA-1-1 can significantly reduce the sensitivity of different cancer cells to cisplatin, and the IC50 values ​​of cisplatin are reduced by 26.5%, 39.1%, 41.9%, 29.1% and 47.6%, respectively, indicating that HCA-1-1 has good anti-cancer sensitization activity.

[0122] Example 2

[0123] This embodiment provides a novel method for preparing a dihydropyridine derivative, and its synthetic route is as follows:

[0124] The specific steps are as follows:

[0125] Under N2 conditions, 9-fluorenone (10.9 mmol, 1.9 g) and 3-hydroxy-4-aminobenzylamine (13.08 mmol, 1.8 g) were added to a 250 mL three-necked flask, and 100 mL of toluene was added to completely dissolve the mixture. After reacting at room temperature for 1 h, ethyl acetoacetate (21.8 mmol, 2.84 g) and ammonium acetate (21.8 mmol, 1.68 mg) were added and refluxed for 20 h. After the reaction was complete, the mixture was evaporated under reduced pressure and purified by column chromatography to obtain Intermediate I.

[0126] To a 100 mL round-bottom flask, add Intermediate I (5.28 mmol, 1.9 g) and sodium hydroxide (5.28 mmol, 211 mg). Add 30 mL of water to completely dissolve the mixture. Saturated chloroacetic acid solution (7.92 mmol, 7.92 mL) and 10 mL of saturated sodium hydroxide solution are then added dropwise at room temperature, controlling the addition rate to maintain the pH between 9 and 10. Heat to 80-90°C and react for 3 hours. After the reaction is complete, adjust the mixture to pH 1 with hydrochloric acid, filter the mixture, wash with water, and dry, then recrystallize from petroleum ether to obtain Intermediate II.

[0127] To a 250 mL round-bottom flask, intermediate II (5.28 mmol, 2.21 g) and 2-methoxy-5-aminophenol (6.34 mmol, 875 mg) were added. 100 mL of anhydrous dichloromethane was then added to completely dissolve the mixture. N,N-dicyclohexylcarbodiimide (6.34 mmol, 1.3 g) and 4-dimethylaminopyridine (0.264 mmol, 38 mg) were then added and allowed to react at room temperature for 18 hours. After the reaction was complete, the mixture was filtered and the filtrate was extracted with dichloromethane. The organic layer was washed sequentially with dilute hydrochloric acid, saturated aqueous sodium bicarbonate solution, and water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain HCA-2-1.

[0128] The HCA-2-1 compound prepared in Example 2 is a specific form of the HCA-2 compound, that is, in the HCA-2 compound, R2 is -CH2CH3 and R5 is -OH.

[0129] The H NMR characterization data of the dihydropyridine derivative (HCA-2-1 compound) prepared in Example 2 are as follows: 1H NMR (500 MHz, Chloroform-d) δ 9.34 (s, 1H), 8.56 (s, 1H), 7.34 (s, 1H), 7.76 (d, J = 1.5 Hz, 1H), 7.09 (dd, J = 7.5, 1.5 Hz, 1H), 6.88-6.81 (m, 3H), 6.79(d,J=7.3Hz,1H),4.88(q,J=1.4Hz,1H),4.79(s,2H),4.53(d,J=7.0Hz,1H),4.32(d,J=7.0 Hz, 1H), 4.00 (q, J = 8.0Hz, 4H), 3.76 (s, 2H), 2.27 (dd, J = 2.2, 0.9Hz, 6H), 1.21 (t, J = 8.0Hz, 6H).

[0130] The characterization data of HCA-2-1 compound are as follows: 13C NMR (125MHz, Chloroform-d) δ168.74, 168.65, 167.53, 167.31, 166.98, 166.97, 166.96, 147.36, 147.12, 147.08, 147.07, 145.98, 145.95, 145.79, 145.77, 145.74, 145.72, 144.93, 144.91, 144.87, 144.89, 137.09, 137.37, 136.50, 136.45, 135.84, 119.84, 119.82, 119.79 This confirmed that the target product was HCA-2-1.

[0131] Example 3

[0132] This embodiment provides a novel method for preparing a dihydropyridine derivative, and its synthetic route is as follows:

[0133] The specific steps are as follows:

[0134] 1) To a 250 mL round-bottom flask, 4-hydroxy-3-nitrobenzaldehyde (15 mmol, 2.5 g), ethyl propionyl acetate (21 mmol, 3.02 g), and ammonium acetate (21 mmol, 1.6 g) were added, and 180 mL of ethanol was added to completely dissolve the mixture. The reaction was refluxed for 18 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, recrystallized from ethanol, and filtered and dried to obtain Intermediate A;

[0135] 2) To a 250 mL round-bottom flask, Intermediate A (10.9 mmol, 4.2 g), bromopropyl alcohol (21.8 mmol, 3 g), and potassium carbonate (21.8 mmol, 3.02 g) were added, followed by complete dissolution in 40 mL of N,N-dimethylformamide. The mixture was heated to 100°C and reacted for 8 h. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the pH of the mixture was adjusted to neutral with hydrochloric acid. The mixture was extracted with ethyl acetate, and the organic layer was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain Intermediate B.

[0136] 3) To a 250 mL round-bottom flask, intermediate B (2.36 mmol, 5.29 g), 2-iodoacylbenzoic acid (18.88 mmol, 5.29 g), and iodine (3.78 mmol, 958 mg) were added, and 15 mL of N,N-dimethylformamide was added to completely dissolve the mixture. The mixture was heated to 100° C. and reacted for 18 h. After the reaction was complete, the reaction mixture was cooled to room temperature, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and separated by column chromatography to obtain intermediate C.

[0137] 4) To a 250 mL round-bottom flask, intermediate C (5.34 mmol, 2.46 g) and 3-amino-5-fluorotrifluoromethylbenzene (6.41 mmol, 1.15 g) were added, followed by complete dissolution with anhydrous dichloromethane. N,N-dicyclohexylcarbodiimide (6.41 mmol, 1.3 g) and 4-dimethylaminopyridine (0.32 mmol, 33 mg) were then added and allowed to react at room temperature for 18 h. After completion of the reaction, the mixture was filtered and the filtrate was extracted with dichloromethane. The organic layer was washed sequentially with dilute hydrochloric acid, saturated aqueous sodium bicarbonate solution, and water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain HCA-3.

[0138] The HCA-3-1 compound prepared in Example 3 is a specific form of the HCA-3 compound, that is, in the HCA-3 compound, R1 is -CH2CH3 and R6 is -F.

[0139] The H NMR characterization data of the dihydropyridine derivative (HCA-3-1 compound) prepared in Example 3 are as follows: 1H NMR (500 MHz, Chloroform-d) δ 9.47 (s, 1H), 9.93 (d, J = 6.3 Hz,, 1H), 8.67 (t, J = 1.5 Hz, 1H), 7.94-7.89 (m, 1H), 7.49-7.43 (m, 2H), 7.22-7.17 (m, 3H), 5.06-5.20 (m, 1H), 4.81(d,J=12.4Hz,1H),4.71(d,J=12.3Hz,1H),4.17(qd,J=7.9,4.0Hz,2H),4.10-3.92(m,2H),2.75(dqd,J= 12.3, 8.0, 0.9Hz, 1H), 2.68 (dqd, J=12.5, 8.0, 0.9Hz, 1H), 1.22 (td, J=8.0, 0.7Hz, 6H), 1.09 (t, J=8.0Hz, 3H).

[0140] The characterization data of the carbon nuclear magnetic resonance spectrum of the HCA-3-1 compound are as follows: 13C NMR (125MHz, Chloroform-d) δ168.95, 168.55, 167.36, 167.33, 167.32, 167.29, 166.95, 166.92, 163.89, 163.87, 163.85, 161.87, 161.86, 161.84, 150.57, 150.54, 150.51, 150.41, 150.17, 1 39.56,139.55,139.50,139.48,138.94,138.06,135.21,135.64,135.47,135.45,135.44,135.41,135.40,135.39,132.21,132.18,132.33,132.45,132.44,132.08,127.05,127.04,127.03, 127.02,127.00,126.99,126.97,126.97,124.87,124.56,122.67,122.02,116.43,116.41,116.39,116.36,116.34,113.24,113.22,113.19,113.16,113.14,113.11,109.27,109.25,109.2 2,109.20,109.11,109.09,109.06,109.04,100.49,100.15,100.13,68.35,68.31,62.16,62.14,62.11,60.53,60.50,36.23,36.20,36.17,36.13,36.10,23.35,23.33,14.44,14.41,14.28,

[0141] 14.25, 12.89, 12.87. This confirmed that it was indeed the target product HCA-3-1.

[0142] The present application has been described in detail above. Specific examples have been used herein to illustrate the principles and implementation methods of the present application. The description of the above embodiments is intended only to facilitate understanding of the present application and its core concepts. It should be noted that, without departing from the principles of the present application, a number of improvements and modifications may be made to the present application by a person skilled in the art, and such improvements and modifications shall fall within the scope of protection of the claims of the present application.

Claims

1. A dihydropyridine derivative, a pharmaceutically acceptable salt or a stereoisomer thereof, characterized in that: The structure of the dihydropyridine derivative is shown in the following formula (I): Wherein, R1 is selected from any one of -H, -CH3, -CH2CH3, and -OCH3; R2 is selected from any one of -H, -CH3, -CH2CH3; R3 is selected from any one of -H, -OH, -Br, and -NH2; R4 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OH, -NH2; R5 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OCH3, -NH2; R6 is selected from any one of -H, -CH3, -Br, -F, -CF3, -NH2; R7 is selected from any one of -H, -CH3, -NO2, and -NH2.

2. The dihydropyridine derivative, pharmaceutically acceptable salt or stereoisomer thereof according to claim 1, characterized in that: The structure of the dihydropyridine derivative is shown in the following formula (II): Wherein, R1 is selected from any one of -H, -CH3, -CH2CH3, and -OCH3; R5 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OCH3, and -NH2.

3. The dihydropyridine derivative, pharmaceutically acceptable salt or stereoisomer thereof according to claim 1, characterized in that: The structure of the dihydropyridine derivative is shown in the following formula (III): Wherein, R2 is selected from any one of -H, -CH3, -CH2CH3; R4 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OH, and -NH2.

4. The dihydropyridine derivative, pharmaceutically acceptable salt or stereoisomer thereof according to claim 1, characterized in that: The structure of the dihydropyridine derivative is shown in the following formula (IV): Wherein, R1 is any one of -H, -CH3, and -OCH3; R6 is selected from any one of -H, -CH3, -Br, -F, -CF3, and -NH2.

5. A method for preparing a dihydropyridine derivative, characterized in that: Including at least one of methods 1 to 3, wherein: Method 1 comprises the following steps: Wherein, R1 is any one of -H, -CH3, and -OCH3; R5 is any one of -H, -CH3, -Br, -Cl, -CF3, -OCH3, -NH2; 4-Hydroxy-3-nitrobenzaldehyde first reacts with ammonium acetate and β-keto acid to produce intermediate 1, which is then dissolved in aqueous sodium hydroxide solution, followed by dropwise addition of chloroacetic acid and a saturated sodium hydroxide solution to produce intermediate 2. Intermediate 2 is then dissolved in methanol, followed by addition of aqueous sodium hydroxide solution to produce intermediate 3. The prepared intermediate 3 and the corresponding aniline compound are then dissolved in dichloromethane, followed by addition of N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine to produce HCA-1. Method 2 includes the following steps: Wherein, R2 is selected from any one of -H, -CH3, -CH2CH3; R4 is selected from any one of -H, -CH3, -Br, -Cl, -CF3, -OH, -NH2; 9-Fluorenone and 3-hydroxy-4-aminobenzylamine are first reacted with β-ketoester to prepare intermediate 4, which is then dissolved in water with sodium hydroxide and reacted with chloroacetic acid to prepare intermediate 5. Intermediate 5 and the corresponding amine are dissolved in dichloromethane, and N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine are added to react to obtain HCA-2 compound. Method 3 comprises the following steps: Wherein, R1 is selected from any one of -H, -CH3, -CH2CH3, and -OCH3; R6 is selected from any one of -H, -CH3, -Br, -F, -CF3, -NH2; 4-Hydroxy-3-nitrobenzaldehyde is first reacted with β-ketoester and ammonium acetate to prepare intermediate 1, which is then dissolved in N,N-dimethylformamide with bromopropyl alcohol and potassium carbonate in appropriate proportions to prepare intermediate 6. Intermediate 6, 2-iodoacylbenzoic acid and iodine are then dissolved in N,N-dimethylformamide to react to prepare intermediate 7. Intermediate 7 and the corresponding aniline derivative are then dissolved in dichloromethane, and N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine are added to react to obtain HCA-3 compound.

6. The preparation method according to claim 5, characterized in that The specific steps of the method 1 are as follows: 1) 4-hydroxy-3-nitrobenzaldehyde, β-ketoacid, and ammonium acetate are mixed in appropriate proportions and dissolved in ethanol, and refluxed for 16-18 hours. After the reaction is complete, the mixture is cooled, filtered, and recrystallized from ethanol. The solid is filtered and dried to obtain intermediate 1, wherein the molar ratio of 4-hydroxy-3-nitrobenzaldehyde, β-ketoacid, and ammonium acetate is 1-1.5:2-4:2-4. 2) Dissolve intermediate 1 and ethyl chloroacetate in acetonitrile, add potassium iodide and anhydrous potassium carbonate, and react at 100°C for 8 hours. After the reaction is complete, cool to room temperature, filter, wash the solid with ethyl acetate, extract the filtrate with brine and ethyl acetate, dry the organic layer over anhydrous sodium sulfate, concentrate, and purify by column chromatography to obtain intermediate 2; wherein the molar ratio of intermediate 1, ethyl chloroacetate, potassium iodide, and anhydrous potassium carbonate is 1:1-2:0.5:3; 3) Dissolve intermediate 2 in methanol, add 6 mol / L sodium hydroxide aqueous solution, and reflux for 8 hours; after the reaction is complete, concentrate the solvent under reduced pressure, adjust the pH to 2 with 6 mol / L hydrochloric acid aqueous solution, extract with dichloromethane, dry over anhydrous sodium sulfate, concentrate, and recrystallize from ethanol to obtain intermediate 3; 4) Dissolving intermediate 3 and its corresponding aniline derivative in anhydrous dichloromethane, adding N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine, and reacting at room temperature for 18 hours; after the reaction is complete, filtering, extracting the filtrate with dichloromethane, and washing the organic layer with dilute hydrochloric acid, saturated sodium bicarbonate aqueous solution, and water, drying over anhydrous sodium sulfate, concentrating, and purifying by column chromatography to obtain HCA-1 compound; wherein the molar ratio of intermediate 3, the corresponding aniline derivative, N,N-dicyclohexylcarbodiimide, and 4-dimethylaminopyridine is 1:1-1.5:1-2:0.

2.

7. The preparation method according to claim 5, characterized in that The specific steps of method 2 are as follows: 1) Under N2 conditions, 3-hydroxy-4-aminobenzylamine was added to a toluene solution of 9-fluorenone, and the mixture was stirred at room temperature for 1 hour. β-keto acid and ammonium acetate were then added, and the mixture was refluxed for 20 hours. After the reaction was complete, the mixture was evaporated under reduced pressure and purified by column chromatography to obtain intermediate 4; wherein the molar ratio of 9-fluorenone, 3-hydroxy-4-aminobenzylamine, β-keto acid, and ammonium acetate was 0.5:1:1-2:1-2; 2) Dissolve intermediate 4 and sodium hydroxide in water, and add saturated chloroacetic acid solution and saturated sodium hydroxide solution dropwise in sequence at room temperature, controlling the addition rate to maintain the pH value at 9-10; heat to 80-90°C and react for 3 hours; after the reaction is complete, adjust the pH of the mixture to 1 with hydrochloric acid, filter the mixture, wash with water, dry, and then recrystallize from petroleum ether to obtain intermediate 5; wherein the molar ratio of intermediate 4, sodium hydroxide, and chloroacetic acid is 1:1:1.5; 3) Dissolving intermediate 5 and its corresponding aniline derivative in anhydrous dichloromethane, adding N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine, and reacting at room temperature for 18 hours; after the reaction is complete, filtering, extracting the filtrate with dichloromethane, and washing the organic layer with dilute hydrochloric acid, saturated sodium bicarbonate aqueous solution, and water, drying over anhydrous sodium sulfate, concentrating, and purifying by column chromatography to obtain HCA-2 compound; wherein the molar ratio of intermediate 5, the corresponding aniline derivative, N,N-dicyclohexylcarbodiimide, and 4-dimethylaminopyridine is 1:1-1.5:1-2:0.

2.

8. The preparation method according to claim 5, characterized in that The specific steps of method 3 are as follows: 1) 4-hydroxy-3-nitrobenzaldehyde, β-ketoester, and ammonium acetate are mixed in appropriate proportions and dissolved in ethanol, and refluxed for 16-18 hours. After the reaction is complete, the mixture is cooled, filtered, and recrystallized from ethanol. The solid is filtered and dried to obtain intermediate 1, wherein the molar ratio of 4-hydroxy-3-nitrobenzaldehyde, β-ketoacid, and ammonium acetate is 1-1.5:2-4:2-4; 2) Dissolve intermediate 1, bromopropanol, and potassium carbonate in N,N-dimethylformamide and react at 80-100°C for 8 hours. After the reaction is complete, cool to room temperature. After filtering, adjust the pH of the mixture to neutral with hydrochloric acid, extract with ethyl acetate, and dry the organic layer over anhydrous sodium sulfate. Concentrate and purify by column chromatography to obtain intermediate 6. The molar ratio of intermediate 1 to bromopropanol and potassium carbonate is 1:1-2:1-2. 3) Dissolving intermediate 6, 2-iodoacetylbenzoic acid, and iodine in N,N-dimethylformamide and heating at 100°C for 18 hours. After the reaction is complete, the reaction mixture is cooled to room temperature, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and separated by column chromatography to obtain intermediate 7; wherein the molar ratio of intermediate 6, 2-iodoacetylbenzoic acid, and iodine is 0.25:2:0.4; 4) Dissolving intermediate 7 and its corresponding aniline derivative in anhydrous dichloromethane, adding N,N-dicyclohexylcarbodiimide and 4-dimethylaminopyridine, and reacting at room temperature for 18 hours; after the reaction is complete, filtering, extracting the filtrate with dichloromethane, and washing the organic layer with dilute hydrochloric acid, saturated sodium bicarbonate aqueous solution, and water, drying over anhydrous sodium sulfate, concentrating, and purifying by column chromatography to obtain HCA-3; wherein the molar ratio of intermediate 7, the corresponding aniline derivative, N,N-dicyclohexylcarbodiimide, and 4-dimethylaminopyridine is 1:1-1.5:1-2:0.

2.

9. An intermediate for preparing the dihydropyridine derivative, a pharmaceutically acceptable salt or stereoisomer thereof according to any one of claims 2 to 4, characterized in that: The intermediate is at least one of the following intermediates 1-7; Wherein, R1 is selected from any one of -H, -CH3, -CH2CH3, and -OCH3; R2 is selected from any one of -H, -CH3, and -CH2CH3.

10. A pharmaceutical composition, characterized in that The invention comprises the dihydropyridine derivative according to any one of claims 1 to 4, a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier, excipient or diluent.

11. Use of the dihydropyridine derivative according to any one of claims 1 to 4, a pharmaceutically acceptable salt or stereoisomer thereof, or the pharmaceutical composition according to claim 10 in the preparation of a drug for treating or preventing hypertension combined with cancer.

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