Hederagenin C-23 heterocyclic ester derivative as well as preparation method and application thereof

Abstract

The invention discloses a hederagenin C-23 heterocyclic ester derivative as well as a preparation method and application thereof, and belongs to the technical field of organic chemistry. The structure of the derivative is shown in the specification, wherein R represents pyrazole, thiazole, oxazole, furan, pyrimidine, pyrazine or indole. Pharmacological tests show that part of hederagenin C-23 heterocyclic ester derivatives (T-BZ, T-SZ, T-EZ, T-MD and TBQ) prepared by the invention have better MDR reversal activity than verapamil, can significantly increase the sensitivity (p < 0.0001) of drug-resistant KBV cells to paclitaxel, are P-gp inhibitors with more excellent MDR reversal activity, can be used in combination with common antitumor drugs, and can be used for preparing drugs for treating tumors. The good anti-tumor activity is realized.

Description

Technical Field

[0001] This invention relates to hederone derivatives, their preparation methods and applications, specifically to hederone C-23 heterocyclic ester derivatives, their preparation methods and applications, belonging to the field of organic chemistry technology. Background Technology

[0002] The incidence and mortality rates of malignant tumors worldwide are on a continuous upward trend, becoming a major public health problem that seriously threatens human health. According to the latest statistics, due to the combined effects of population aging and overall growth, it is estimated that by 2050, the number of new malignant tumor cases worldwide will exceed 33 million, and the number of deaths will exceed 18 million, making the prevention and control situation extremely challenging. Currently, among tumor treatment methods such as radiotherapy, surgery, immunotherapy, and chemotherapy, chemotherapy is the most important method for treating malignant tumors. However, its efficacy is often limited by drug resistance. Multidrug resistance (MDR) has become a key factor affecting the success of chemotherapy. MDR refers to the cross-resistance of tumor cells to multiple chemotherapeutic drugs with different structures and mechanisms of action, directly leading to more than 90% of chemotherapy failures and exacerbating the risk of tumor recurrence, metastasis, and patient death. It has become a critical problem that urgently needs to be solved in the field of tumor chemotherapy. P-glycoprotein (P-gp), as the most representative MDR-related transmembrane transporter, is the core target of current MDR inhibitor research. In tumor cells, overexpressed P-gp uses the energy released by ATP hydrolysis to pump various chemotherapeutic drugs out of the tumor cells, reducing intracellular drug concentration and leading to MDR and chemotherapy failure. Inhibiting P-gp efflux can significantly increase the sensitivity of drug-resistant tumor cells to chemotherapeutic drugs. Therefore, designing and synthesizing highly effective P-gp inhibitors is an effective strategy to overcome MDR. Currently used clinical P-gp inhibitors (such as verapamil, bilicolda, and taribineda) have significant limitations: some P-gp inhibitors have high self-toxicity or may interfere with the normal metabolic processes of other drugs, and may exacerbate toxic side effects when used in combination with chemotherapeutic drugs. As a result, all three generations of P-gp inhibitors have failed to pass clinical trials and achieve widespread application. It has been reported that various structural types of natural products and their derivatives have been proven to have MDR reversal activity, making natural products one of the important sources of P-gp inhibitors.

[0003] Hederagenin (H) is an oleanane-type pentacyclic triterpenoid compound, mainly extracted from plants of the Araliaceae, Fabaceae, and Vitaceae families. It possesses various pharmacological activities, including anti-inflammatory, antioxidant, and anti-tumor proliferation effects. Our research group previously used hederagenin as a lead compound and modified its structure at the C-3, C-23, and C-28 positions. Our studies revealed that hederagenin derivatives HBQ, HBQ-5, HBQ-P8, and H628 all exhibit certain in vitro and in vivo MDR reversal activities, and the mechanism of action has been preliminarily elucidated (H6, a novel hederagenin derivative, reverses multidrug resistance in vitro and in vivo, Yang YT, Guan DK, Lei L, et al., Toxicology & Applied Pharmacology, 2018, 341: 98-105). Therefore, structural modification of hederogens to enrich the structural types of hederogen derivatives and the development of novel hederogen P-gp inhibitors are the focus of future research. Summary of the Invention

[0004] The purpose of this invention is to provide a novel hederaponin derivative with superior MDR reversal activity, as well as a method for preparing the derivative and its applications.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: The structure of the heterocyclic ester derivative at the C-23 position of hederasein is shown below:

[0006] Where R represents c.

[0007] Preferably, the aforementioned hederonein C-23 heterocyclic ester derivative is: 3-Carbonyl-23-((1H-pyrazole-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((thiazolyl-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((oxazol-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((furan-3-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((pyrimidin-2-carbonyl)oxy)-oleanolic-12-en-28-benzyl ester; 3-Carbonyl-23-((pyrazin-2-carbonyl)oxy)-oleanolic-12-en-28-benzyl ester; Alternatively, 3-carbonyl-23-((1H-indole-2-carbonyl)oxy)-oleanolic-12-ene-28-benzyl ester.

[0008] The preparation method of the aforementioned hederonein C-23 heterocyclic ester derivative includes the following steps: (1) Using ivy saponin as raw material, in the presence of inorganic base, benzyl bromide protects the carboxyl group at C-28 of ivy saponin; (2) Tert-butyldimethylchlorosilane protects the C-23 hydroxyl group of ivy saponin; (3) Oxidation of the C-3 hydroxyl group of ivy saponin with pyridine chlorochromate; (4) Remove tert-butyldimethylchlorosilane under hydrochloric acid and acetone conditions; (5) Under the catalysis of 4-dimethylaminopyridine and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, the C-23 hydroxyl group of iposidegenin reacts with 1H-pyrazole-4-carboxylic acid, thiazole-4-carboxylic acid, oxazole-4-carboxylic acid, furan-3-carboxylic acid, pyrimidine-2-carboxylic acid, pyrazine-2-carboxylic acid or indole-2-carboxylic acid to give the crude product; (6) The crude product was purified by column chromatography to obtain the target compound.

[0009] Preferably, the preparation method of the aforementioned hederonein C-23 heterocyclic ester derivative specifically includes the following steps: (1) Dissolve ivy saponin in N,N-dimethylformamide, add inorganic base and benzyl bromide, stir in a water bath at 50°C, and purify the product after the reaction to obtain intermediate product H1; (2) Dissolve H1 in dichloromethane, add 4-dimethylaminopyridine and tert-butyldimethylchlorosilane, stir at room temperature, and purify the product after the reaction is complete to obtain intermediate product H2; (3) Dissolve H2 in dichloromethane, add freshly prepared pyridine chlorochromate, stir at room temperature, and purify the product after the reaction is complete to obtain intermediate product H3; (4) Dissolve H3 in acetone, then add hydrochloric acid, stir at room temperature, and purify the product after the reaction is complete to obtain intermediate product H4; (5) Dissolve H4 in dichloromethane, and add dimethylaminopyridine, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and reactants in sequence. Stir at room temperature. After the reaction is completed, dilute, wash and dry the product in sequence to obtain crude product. The reactants are 1H-pyrazole-4-carboxylic acid, thiazole-4-carboxylic acid, oxazole-4-carboxylic acid, furan-3-carboxylic acid, pyrimidine-2-carboxylic acid, pyrazine-2-carboxylic acid or indole-2-carboxylic acid. (6) The crude product was purified by silica gel column chromatography to obtain the target compound.

[0010] The aforementioned application of the hederonein C-23 heterocyclic ester derivative in the preparation of P-gp inhibitors, wherein the hederonein C-23 heterocyclic ester derivative is: 3-Carbonyl-23-((1H-pyrazole-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((thiazolyl-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((oxazol-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((pyrimidin-2-carbonyl)oxy)-oleanolic-12-en-28-benzyl ester; Alternatively, 3-carbonyl-23-((pyrazin-2-carbonyl)oxy)-oleanolic-12-ene-28-benzyl ester.

[0011] The advantages of this invention are as follows: This invention introduces heterocyclic structures such as pyrazole, thiazole, oxazole, furan, pyrimidine, pyrazine, and indole into the C-23 hydroxyl group of hederogenin to prepare hederogenin C-23 heterocyclic ester derivatives. Pharmacological tests show that some of the hederogenin C-23 heterocyclic ester derivatives (T-BZ, T-SZ, T-EZ, T-MD, TBQ) prepared in this invention have better MDR reversal activity than verapamil and can significantly increase the sensitivity of drug-resistant KBV cells to paclitaxel (p<0.0001). They are P-gp inhibitors with better MDR reversal activity and can be used in combination with commonly used antitumor drugs to exert good antitumor activity. Attached Figure Description

[0012] Figure 1 This is a graph showing the results of the analysis of significant differences in the survival rate of drug-resistant KBV cells under the combined use of each compound and paclitaxel (paclitaxel-containing treatment group). **** indicates p<0.0001. Detailed Implementation

[0013] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0014] I. Structure of hederone derivatives The hederone derivative provided by this invention is a hederone derivative of a heterocyclic ester modified at the C-23 position, and its structure is shown below:

[0015] Where R represents pyrazole, thiazole, oxazole, furan, pyrimidine, pyrazine, or indole.

[0016] Some heterocyclic ester derivatives of hederonegenin at position C-23 are: 3-Carbonyl-23-((1H-pyrazole-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((thiazolyl-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((oxazol-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((furan-3-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((pyrimidin-2-carbonyl)oxy)-oleanolic-12-en-28-benzyl ester; 3-Carbonyl-23-((pyrazin-2-carbonyl)oxy)-oleanolic-12-en-28-benzyl ester; Alternatively, 3-carbonyl-23-((1H-indole-2-carbonyl)oxy)-oleanolic-12-ene-28-benzyl ester.

[0017] II. Preparation methods of ivy saponin derivatives

[0018] The preparation method of the above-mentioned ivy saponin derivatives provided by the present invention includes the following steps: (1) Using ivy saponin as raw material, in the presence of inorganic base, benzyl bromide protects the carboxyl group at C-28 of ivy saponin; (2) Tert-butyldimethylchlorosilane protects the C-23 hydroxyl group of ivy saponin; (3) Pyridine chlorochromate oxidizes the C-3 hydroxyl group of ivy saponin; (4) Remove tert-butyldimethylchlorosilane under hydrochloric acid and acetone conditions; (5) Under the catalysis of 4-dimethylaminopyridine (DMAP) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), the C-23 hydroxyl group of iposidegenin reacts with 1H-pyrazole-4-carboxylic acid, thiazole-4-carboxylic acid, oxazole-4-carboxylic acid, furan-3-carboxylic acid, pyrimidine-2-carboxylic acid, pyrazine-2-carboxylic acid or indole-2-carboxylic acid to give the crude product; (6) The crude product was purified by column chromatography to obtain the target compound. Example 1

[0019] The raw material, ivy saponin (1.0 mmol), was dissolved in N,N-dimethylformamide (DMF, 15.0 mL), and potassium carbonate (2.1 mmol) and benzyl bromide (1.3 mmol) (carboxyl protecting agent) were added. The mixture was stirred in a water bath at 50 °C for 8 h. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with ethyl acetate (20.0 mL), washed three times with water, and then washed twice with saturated brine. The solution was then dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum distillation. Finally, the solution was subjected to silica gel column chromatography (V... 石油醚 V 乙酸乙酯 =10:1), and each elution component was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white solid (yield 83.0%), denoted as H1.

[0020] H1 (0.8 mmol) was dissolved in dichloromethane (20.0 mL), and 4-dimethylaminopyridine (DMAP, 1.0 mmol), a catalyst and acid-binding agent, and tert-butyldimethylchlorosilane (TBSCl, 2.4 mmol), a (C-23) hydroxyl protecting agent, were added. The mixture was stirred at room temperature for 6 h. After the reaction was complete, the solvent was first evaporated, then diluted with ethyl acetate (20.0 mL), followed by washing with 5% (w / v) hydrochloric acid until acidic, then washing with saturated brine until neutral, drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure. Finally, the solution was analyzed by silica gel column chromatography (V... 石油醚 V 乙酸乙酯 =30:1), and each elution component was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white solid (yield 70.0%), denoted as H2.

[0021] H2 (0.6 mmol) was dissolved in dichloromethane (15.0 mL), and freshly prepared pyridine chlorochromate (1.3 mmol) as the (C-3 position) hydroxyl oxidant was added. The mixture was stirred at room temperature for 8 h. After the reaction was complete, the solvent was evaporated, and the solution was diluted with ethyl acetate (20.0 mL). The solution was then washed three times with water, followed by washing with saturated brine until neutral. The solution was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and finally analyzed by silica gel column chromatography (V). 石油醚 V 乙酸乙酯 =35:1), and each elution component was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white solid (yield 84.0%), denoted as H3.

[0022] H3 (2.23 mmol) was dissolved in acetone (30.0 mL), and then 10% (w / v) hydrochloric acid (7.0 mL) was added. The mixture was stirred at room temperature for 10 h. After the reaction was complete, the solution was concentrated under reduced pressure, diluted with ethyl acetate (20.0 mL), washed twice with water, then twice with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and finally analyzed by silica gel column chromatography (V). 石油醚 V 乙酸乙酯 =6:1), and each elution component was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white solid (yield 84.5%), denoted as H4.

[0023] H4 (0.089 mmol) was dissolved in dichloromethane (5.0 mL), and the catalyst DMAP (0.27 mmol), condensing agent EDCI (0.45 mmol), and reactant 1H-pyrazole-4-carboxylic acid (0.45 mmol) were added sequentially. The mixture was stirred at room temperature for 3 h. After the reaction was complete, the solution was diluted with ethyl acetate (15.0 mL), and then washed twice each with water, 10% (w / v) hydrochloric acid, and saturated brine. The solution was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to silica gel column chromatography (V... 石油醚 V 乙酸乙酯 =6:1), and each elution fraction was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white oily substance (yield 82.62%), denoted as T-BZ (target compound).

[0024] T-BZ 1 H NMR, 13 The following are the C NMR and HRMS (ESI) data: 1H NMR (400MHz, CDCl3) δ 7.81 (d, J=2.3Hz, 1H, H-pyrazole), 7.34 (d, J=3.6Hz, 5H, H-Ar), 6.71 (d, J=2.3Hz, 1H, H-pyrazole), 5.32-5.29 (m, 1H, H-12), 5.12-5.02 (m, 2H, H-CH2Ar), 4.43-4.28 (m, 2H, H-23), 2.94-2.87 (m, 1H, H-1 8), 2.61-2.39 (m, 2H, H-CH2), 1.92 (dt, J=11.4, 3.9Hz, 4H, H-CH2), 1.75-1.12 (m, 17H, H-CH, H-CH2), 1.08 (s, 3H, CH3), 1.04 (d, J=3.2Hz, 6H, 2×CH3), 0.91 (s, 3H, CH3), 0.88 (s, 3H, CH3), 0.66 (s, 3H, CH3).

[0025] 13 C NMR (101MHz, CDCl3) δ 214.54, 177.54, 161.81, 143.95, 141.32, 136.53, 132.69, 128.55 (2C), 128 .14 ​​(2C), 128.07, 122.25, 107.55, 67.95, 66.07, 50.60, 48.60, 46.90, 46.79 , 45.89, 41.94, 41.65, 39.34, 37.97, 36.46, 35.04, 33.98, 33.20, 32.42, 32.15, 30.80, 27.65, 25.67, 23.73, 23.60, 23.15, 19.71, 17.69, 17.03, 14.99.

[0026] HR-MS (ESI): m / z C 41 H 54 N₂O₅[M+Na] + The calculated value is 677.3925, and the theoretical value is 677.3909.

[0027] T-BZ is: 3-carbonyl-23-((1H-pyrazole-4-carbonyl)oxy)-oleanolic-12-ene-28-benzyl ester. Example 2

[0028] H4 was prepared using ivy saponin as a raw material according to the method in Example 1.

[0029] H4 (0.089 mmol) was dissolved in dichloromethane (5.0 mL), and the catalyst DMAP (0.27 mmol), condensing agent EDCI (0.45 mmol), and reactant thiazole-4-carboxylic acid (0.45 mmol) were added sequentially. The mixture was stirred at room temperature for 3 h. After the reaction was complete, the solution was diluted with ethyl acetate (15.0 mL), and then washed twice each with water, 10% (w / v) hydrochloric acid, and saturated brine. The solution was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to silica gel column chromatography (V... 石油醚 V 乙酸乙酯 =6:1), and each elution fraction was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white oily substance (yield 84.30%), denoted as T-SZ (target compound).

[0030] T-SZ 1 H NMR, 13 The following are the C NMR and HRMS (ESI) data: 1 H NMR (400MHz, CDCl3) δ 8.82 (d, J=2.1Hz, 1H, H-thiazole), 8.10 (d, J=2.1Hz, 1H, H-thiazole), 7.33 (s, 5H, H-Ar), 5.30 ( t, J=3.7Hz, 1H, H-12), 5.11-5.02 (m, 2H, H-CH2Ar), 4.35 (d, J=0.9Hz, 2H, H-23), 2.94-2.86 (m, 1H, H-18), 2.61-2.45 (m, 2H, H-CH2), 1.97-1.88 (m, 4H, H-CH2), 1.73-1.14 (m, 16H, H-CH2), 1.09 (s, 3 H, CH3), 1.05 (s, 3H, CH3), 1.03 (s, 3H, CH3), 0.91 (s, 3H, CH3), 0.88 (s, 3H, CH3), 0.66 (s, 3H, CH3).

[0031] 13C NMR (101MHz, CDCl3) δ 214.54, 177.49, 160.85, 153.57, 147.93, 143.96, 136.51, 128.55 (2C), 128 .15 (2C), 128.07, 127.08, 122.26, 68.53, 66.07, 50.57, 48.82, 46.88, 46.76 , 45.94, 41.96, 41.65, 39.33, 38.08, 36.51, 35.01, 33.97, 33.19, 32.40, 32.12, 30.80, 27.67, 25.69, 23.72, 23.61, 23.14, 19.79, 17.53, 16.99, 15.10.

[0032] HR-MS (ESI): m / z C 41 H 53 NO5S[M+Na] + The calculated value is 694.3537, and the theoretical value is 694.3524.

[0033] T-SZ is: 3-carbonyl-23-((thiazol-4-carbonyl)oxy)-oleanolic-12-ene-28-benzyl ester. Example 3

[0034] H4 was prepared using ivy saponin as a raw material according to the method in Example 1.

[0035] H4 (0.089 mmol) was dissolved in dichloromethane (5.0 mL), and the catalyst DMAP (0.27 mmol), condensing agent EDCI (0.45 mmol), and reactant oxazole-4-carboxylic acid (0.45 mmol) were added sequentially. The mixture was stirred at room temperature for 3 h. After the reaction was complete, the solution was diluted with ethyl acetate (15.0 mL), and then washed twice each with water, 10% (w / v) hydrochloric acid, and saturated brine. The solution was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to silica gel column chromatography (V... 石油醚 V 乙酸乙酯 =6:1), and each elution fraction was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white oily substance (yield 86.55%), denoted as T-EZ (target compound).

[0036] T-EZ 1 H NMR, 13 The following are the C NMR and HRMS (ESI) data: 1H NMR (400MHz, CDCl3) δ 8.15 (d, J=1.0Hz, 1H, H-oxazole), 7.90 (d, J=1.0Hz, 1H, H-oxazole), 7.33 (s, 5H, H-Ar), 5.30 (t , J=3.7Hz, 1H, H-12), 5.12-5.02 (m, 2H, H-CH2Ar), 4.31 (d, J=1.7Hz, 2H, H-23), 2.91 (dd, J=14.0 , 4.6Hz, 1H, H-18), 2.59-2.43 (m, 2H, H-CH2), 1.99-1.87 (m, 4H, H-CH2), 1.70-1.15 (m, 16H, H-CH 2), 1.07 (s, 6H, 2×CH3), 1.03 (s, 3H, CH3), 0.91 (s, 3H, CH3), 0.88 (s, 3H, CH3), 0.66 (s, 3H, CH3).

[0037] 13 C NMR (101MHz, CDCl3) δ 214.43, 177.50, 160.63, 151.63, 143.96, 143.92, 136.52, 133.12, 128.55 ( 2C), 128.15 (2C), 128.07, 122.26, 68.25, 66.08, 50.50, 48.78, 46.89, 46.76 , 45.94, 41.95, 41.65, 39.33, 38.02, 36.50, 34.99, 33.97, 33.20, 32.41, 32.12, 30.80, 27.68, 25.72, 23.72, 23.60, 23.15, 19.76, 17.53, 17.00, 15.07.

[0038] HR-MS (ESI): m / z C 41 H 53 NO6[M+Na] + The calculated value is 678.3765, and the theoretical value is 678.3752.

[0039] T-EZ is: 3-carbonyl-23-((oxazol-4-carbonyl)oxy)-oleanolic-12-ene-28-benzyl ester. Example 4

[0040] H4 was prepared using ivy saponin as a raw material according to the method in Example 1.

[0041] H4 (0.089 mmol) was dissolved in dichloromethane (5.0 mL), and DMAP (0.27 mmol) catalyst, EDCI (0.45 mmol) condensing agent, and furan-3-carboxylic acid (0.45 mmol) reactant were added sequentially. The mixture was stirred at room temperature for 3 h. After the reaction was complete, the solution was diluted with ethyl acetate (15.0 mL), and then washed twice each with water, 10% (w / v) hydrochloric acid, and saturated brine. The solution was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to silica gel column chromatography (V... 石油醚 V 乙酸乙酯 =6:1), and each elution fraction was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white oily substance (yield 81.95%), denoted as T-FN (target compound).

[0042] T-FN 1 H NMR, 13 The following are the C NMR and HRMS (ESI) data: 1 H NMR (400MHz, CDCl3) δ 7.93 (d, J=0.8Hz, 1H, H-oxole), 7.40 (t, J=1.8Hz, 1H, H-oxole), 7.34 (s, 5H, H-Ar), 6.67 (dd, J=1.9, 0.8Hz, 1H , H-oxole), 5.31 (t, J=3.7Hz, 1H, H-12), 5.12-5.02 (m, 2H, H-CH2Ar), 4.29-4.20 (m, 2H, H-23), 2.95-2.87 (m, 1H , H-18), 2.60-2.38 (m, 2H, H-CH2), 1.97-1.89 (m, 4H, H-CH2), 1.72-1.53 ​​(m, 8H, H-CH2), 1.44-1.17 (m, 8H, H-CH 2), 1.07 (s, 3H, CH3), 1.05 (s, 3H, CH3), 1.04 (s, 3H, CH3), 0.91 (s, 3H, CH3), 0.88 (s, 3H, CH3), 0.66 (s, 3H, CH3).

[0043] 13C NMR (101MHz, CDCl3) δ 214.46, 177.50, 162.72, 147.67, 143.96, 136.52, 128.55 (2C), 128.15 (2C), 1 28.07, 122.28, 119.34, 109.77, 67.63, 66.08, 50.56, 48.65, 46.90, 46.81, 45 .93, 41.95, 41.66, 39.34, 38.00, 36.45, 35.02, 33.98, 33.20, 32.41, 32.17, 30.81, 29.81, 27.68, 25.66, 23.72, 23.61, 23.15, 19.70, 17.66, 17.04, 14.96.

[0044] HR-MS (ESI): m / z C 42 H 54 O6[M+Na] + The calculated value is 677.3813, and the theoretical value is 677.3806.

[0045] T-FN is: 3-carbonyl-23-((furan-3-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester. Example 5

[0046] H4 was prepared using ivy saponin as a raw material according to the method in Example 1.

[0047] H4 (0.089 mmol) was dissolved in dichloromethane (5.0 mL), and DMAP (0.27 mmol) catalyst, EDCI (0.45 mmol) condensing agent, and pyrimidine-2-carboxylic acid (0.45 mmol) reactant were added sequentially. The mixture was stirred at room temperature for 3 h. After the reaction was complete, the solution was diluted with ethyl acetate (15.0 mL), and then washed twice each with water, 10% (w / v) hydrochloric acid, and saturated brine. The solution was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to silica gel column chromatography (V... 石油醚 V 乙酸乙酯 =6:1), and each elution fraction was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white oily substance (yield 85.47%), denoted as T-MD, 85.47% (target compound).

[0048] T-MD 1 H NMR, 13 The following are the C NMR and HRMS (ESI) data: 1H NMR (400MHz, CDCl3) δ 8.87 (d, J=4.8Hz, 2H, H-pyrimidine), 7.43 (t, J=4.9Hz, 1H, H-pyrimidine), 7.35-7.29 (m, 5H, H-Ar), 5.31 (t, J=3.7Hz, 1H, H-12), 5.11-5.02 (m, 2H, H-CH2Ar), 4.40 (d, J=1.2Hz, 2H, H-23), 2.94-2.88 (m, 1H, H-18), 2 .61-2.49 (m, 2H, H-CH2), 1.91 (dt, J=17.8, 8.9, 5.3Hz, 5H, H-CH, H-CH2), 1.75-1.15 (m, 15H, H-CH, H-CH2), 1.10 (s, 3H, CH3), 1.07 (s, 3H, CH3), 1.03 (s, 3H, CH3), 0.91 (s, 3H, CH3), 0.88 (s, 3H, CH3), 0.66 (s, 3H, CH3).

[0049] 13 C NMR (101MHz, CDCl3) δ214.78, 177.48, 163.44, 157.83 (2C), 157.00, 143.94, 136.5 0, 128.53 (2C), 128.14 (2C), 128.06, 123.06, 122.33, 69.48, 66.06, 50.50, 48.27, 46.88, 46.57, 45.94, 41.98, 41.65, 39.34, 37.88, 36.49, 35.13, 33.96, 33.19, 32.39, 32.04, 30.79, 27.67, 25.69, 23.71, 23.61, 23.14, 19.75, 17.49, 16.96, 15.09.

[0050] HR-MS (ESI): m / z C 42 H 54 N₂O₅[M+Na] + The calculated value is 689.3925, and the theoretical value is 689.3908.

[0051] T-MD is: 3-carbonyl-23-((pyrimidin-2-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester. Example 6

[0052] H4 was prepared using ivy saponin as a raw material according to the method in Example 1.

[0053] H4 (0.089 mmol) was dissolved in dichloromethane (5.0 mL), and the catalyst DMAP (0.27 mmol), condensing agent EDCI (0.45 mmol), and reactant pyrazine-2-carboxylic acid (0.45 mmol) were added sequentially. The mixture was stirred at room temperature for 3 h. After the reaction was complete, the solution was diluted with ethyl acetate (15.0 mL), and then washed twice each with water, 10% (w / v) hydrochloric acid, and saturated brine. The solution was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to silica gel column chromatography (V... 石油醚 V 乙酸乙酯 =6:1), and each elution fraction was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white oily substance (yield 83.25%), denoted as T-BQ (target compound).

[0054] T-BQ 1 H NMR, 13 The following are the C NMR and HRMS (ESI) data: 1 H NMR (400MHz, CDCl3) δ 9.17 (d, J=1.5Hz, 1H, H-pyrazine), 8.74-8.69 (m, 2H, H-pyrazine), 7.33 (s, 5H, H-Ar), 5.3 1 (t, J=3.6Hz, 1H, H-12), 5.11-5.01 (m, 2H, H-CH2Ar), 4.49-4.33 (m, 2H, H-23), 2.94-2.87 (m , 1H, H-18), 2.64-2.44 (m, 2H, H-CH2), 2.00-1.89 (m, 4H, H-CH2), 1.77-1.18 (m, 16H, H-CH2), 1.11 (s, 3H, CH3), 1.06 (s, 6H, 2×CH3), 0.91 (s, 3H, CH3), 0.88 (s, 3H, CH3), 0.66 (s, 3H, CH3).

[0055] 13C NMR (101MHz, CDCl3) δ 214.20, 177.49, 163.56, 147.68, 146.02, 144.85, 143.92, 143.62, 136.51, 12 8.54 (2C), 128.15 (2C), 128.07, 122.25, 68.96, 66.07, 50.59, 48.84, 46.87, 4 6.85, 45.90, 41.94, 41.63, 39.35, 38.10, 36.54, 34.96, 33.96, 33.19, 32.40, 32.17, 30.80, 27.68, 25.68, 23.71, 23.59, 23.13, 19.71, 17.58, 17.02, 15.06.

[0056] HR-MS (ESI): m / z C 42 H 54 N₂O₅[M+Na] + The calculated value is 689.3925, and the theoretical value is 689.3911.

[0057] T-BQ is: 3-carbonyl-23-((pyrazin-2-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester. Example 7

[0058] H4 was prepared using ivy saponin as a raw material according to the method in Example 1.

[0059] H4 (0.089 mmol) was dissolved in dichloromethane (5 mL), and DMAP (0.27 mmol) catalyst, EDCI (0.45 mmol) condensing agent, and indole-2-carboxylic acid (0.45 mmol) reactant were added sequentially. The mixture was stirred at room temperature for 3 h. After the reaction was complete, the solution was diluted with ethyl acetate (15.0 mL), and then washed twice each with water, 10% (w / v) hydrochloric acid, and saturated brine. The solution was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to silica gel column chromatography (V... 石油醚 V 乙酸乙酯 =6:1), and each elution fraction was monitored by thin-layer chromatography. The eluent containing the product was collected, concentrated under reduced pressure, and dried in a vacuum drying oven at 50°C for 12 h to obtain a white oily substance (yield 70.90%), denoted as T-YD (target compound).

[0060] T-YD 1 H NMR, 13 The following are the C NMR and HRMS (ESI) data: 1H NMR (400MHz, CDCl3) δ 9.04 (s, 1H, H-NH), 7.67 (dd, J=8.1, 1.0Hz, 1H, H-azaindole), 7.42 (dq, J=8.4, 0.9Hz, 1H, H-azaindole), 7.34 (s, 5H, H-Ar), 7.18-7.10 (m, 2H, H-azaindole), 5.31 (d, J=3.6Hz, 1H, H-12), 5.13-5.04 (m, 2H, H-CH2Ar), 4.44 -4.25 (m, 2H, H-23), 2.91 (dd, J=13.6, 4.7Hz, 1H, H-18), 2.67-2.45 (m, 2H, H-1), 1.99-1.27 (m, 21H, H-CH, H-CH 2), 1.10 (s, 3H, CH3), 1.07 (s, 3H, CH3), 1.06 (s, 3H, CH3), 0.92 (s, 3H, CH3), 0.89 (s, 3H, CH3), 0.68 (s, 3H, CH3).

[0061] 13 C NMR (101MHz, CDCl3) δ 214.66, 177.53, 161.73, 143.96, 137.11, 136.52, 128.55, 128.14, 128.06, 127.52, 127.10, 125.59, 122.69, 122.27, 120.98, 112.09, 108.70, 68.07, 66.07, 50.66, 48. 71, 46.89, 46.76, 45.90, 41.95, 41.64, 39.35, 38.01, 36.50, 35.09, 33.96, 33.19, 32.41, 32.15, 30.79, 27.68, 25.68, 23.72, 23.61, 23.15, 19.76, 17.66, 17.03, 15.02.

[0062] HR-MS (ESI): m / z C 46 H 57 NO5[M+Na] + The calculated value is 726.4134, and the theoretical value is 726.4107.

[0063] T-YD is: 3-carbonyl-23-((1H-indole-2-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester. III. Pharmacological Activities of Hederogen Derivatives

[0064] Drug-resistant KBV cells in logarithmic growth phase were digested with trypsin at a concentration of 0.25% (w / v) and then prepared into a solution with a concentration of 2×10⁻⁶. 4 A single-cell suspension of cells per mL.

[0065] The prepared single-cell suspension was seeded at a rate of 2000 cells / well in a 96-well plate (100 μL of single-cell suspension was added to each well) and incubated at 37°C for 24 h.

[0066] After culture, in the paclitaxel-containing group, each well was added with 50 μL of the test compound (verapamil, T-BZ, T-SZ, T-EZ, T-FN, T-MD, T-BQ, or T-YD, all dissolved in dimethyl sulfoxide (DMSO)) solution and 50 μL of paclitaxel solution. The final concentration of the test compound was 10 μM, and the final concentration of paclitaxel was 100 nM. In the paclitaxel-free group, each well was added with 50 μL of the test compound (verapamil, T-BZ, T-SZ, T-EZ, T-FN, T-MD, T-BQ, or T-YD, all dissolved in DMSO) solution and 50 μL of complete culture medium. The final concentration of the test compound was 10 μM. In the control group, each well was added with 100 μL of complete culture medium. DMSO solution, with a final concentration of DMSO <0.1% (v / v), was used. A blank control group (with only 100 μL of complete culture medium in each well, and no cells) was set up to remove the background. Each group had 3 parallel wells and was cultured at 37°C for 72 h.

[0067] After the culture was completed, the supernatant was discarded, and 100 μL of complete medium containing 0.5 mg / mL thiazolyl blue (MTT) was added to each well of each group. The culture was continued at 37°C for 4 h.

[0068] After the culture was completed, the supernatant was discarded, and 150 μL of DMSO was added to each well of each group to dissolve the MTT formazan precipitate. The mixture was then shaken and mixed with a micro-shaker. The optical density (OD) was then measured using a microplate reader at a reference wavelength of 450 nm and a detection wavelength of 570 nm.

[0069] The following formula was used to calculate the survival rate (%) of resistant KBV cells under the action of each compound alone and in combination with paclitaxel: Cell viability (%) = (mean OD value of the treatment group / mean OD value of the control group) × 100% The calculated survival rates (%) of resistant KBV cells under the treatment of each compound alone (in the group without paclitaxel) and in combination with paclitaxel (in the group containing paclitaxel) are shown in Table 1. The results of the analysis of significant differences in the survival rates of resistant KBV cells under the treatment of each compound in combination with paclitaxel (in the group containing paclitaxel) are shown in Table 1. Figure 1 .

[0070] Table 1. Calculation results of drug-resistant KBV cell survival rate (%) in the treatment groups

[0071] From Table 1 and Figure 1 It can be seen that the compounds (T-BZ, T-SZ, T-EZ, T-MD, and T-BQ) prepared in Examples 1, 2, 3, 5, and 6 all have MDR reversal activity, can significantly increase the sensitivity of drug-resistant KBV cells to the anti-tumor drug paclitaxel, and their MDR reversal activity is significantly better than that of the equivalent dose of the positive control drug verapamil (p<0.0001), making them superior P-gp inhibitors to verapamil.

[0072] In summary, the hederone saponin C-23 heterocyclic ester derivatives (especially T-BZ, T-SZ, T-EZ, T-MD and T-BQ) provided by this invention can be used in combination with the commonly used antitumor drug paclitaxel to exert good antitumor activity.

[0073] It should be noted that the above embodiments are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those skilled in the art can make other variations or modifications based on the above description. It is impossible to exhaustively list all possible implementations here. All obvious variations or modifications derived from the technical solutions of this invention are still within the scope of protection of this invention.

Claims

1. A C-23 heterocyclic ester derivative of a saponinogen of Hedera helix, characterized in that, The structure of the C-23 heterocyclic ester derivative of the saponin genin is as follows:

2. Wherein, R represents pyrazole, thiazole, oxazole, furan, pyrimidine, pyrazine or indole.

3. The C-23 heterocyclic ester derivatives of hederagenin according to claim 1, characterized by, The C-23 heterocyclic ester derivative of the saponin genin is: 3-carbonyl-23-((1H-pyrazole-4-carbonyl)oxy)-olean-12-ene-28-oic acid benzyl ester; 3-carbonyl-23-((thiazole-4-carbonyl)oxy)-olean-12-ene-28-oic acid benzyl ester; 3-carbonyl-23-((oxazole-4-carbonyl)oxy)-olean-12-ene-28-oic acid benzyl ester; 3-carbonyl-23-((furan-3-carbonyl)oxy)-olean-12-ene-28-oic acid benzyl ester; 3-carbonyl-23-((pyrimidine-2-carbonyl)oxy)-olean-12-ene-28-oic acid benzyl ester; 3-carbonyl-23-((pyrazine-2-carbonyl)oxy)-olean-12-ene-28-oic acid benzyl ester; Or, 3-carbonyl-23-((1H-indole-2-carbonyl)oxy)-olean-12-ene-28-oic acid benzyl ester.

4. The process for the preparation of the C-23 heterocyclic ester derivatives of hopogenin according to claim 1, characterized by, Comprising the following steps: (1) Using saponin genin as raw material, in the presence of inorganic base, bromobenzyl protects the C-28 carboxyl of saponin genin; (2) Tert-butyl dimethyl chlorosilane protects the C-23 hydroxyl of saponin genin; (3) Pyridine chlorochromate oxidizes the C-3 hydroxyl of saponin genin; (4) Under the condition of hydrochloric acid and acetone, remove tert-butyl dimethyl chlorosilane; (5) Under the catalysis of 4-dimethylaminopyridine and 1-ethyl-(3-dimethylaminopropyl) carbonyldiimidazole hydrochloride, the C-23 hydroxyl of saponin genin reacts with 1H-pyrazole-4-carboxylic acid, thiazole-4-carboxylic acid, oxazole-4-carboxylic acid, furan-3-carboxylic acid, pyrimidine-2-carboxylic acid, pyrazine-2-carboxylic acid or indole-2-carboxylic acid to obtain a crude product; (6) The crude product is purified by column chromatography to obtain the target compound.

5. The process for the preparation of C-23 heterocyclic ester derivatives of hederagenin according to claim 3, characterized by, Specifically comprising the following steps: (1) Dissolve the saponin genin in N, N-dimethylformamide, add inorganic base and bromobenzyl, stir in a 50°C water bath, purify the product after the reaction is completed, and obtain intermediate product H1; (2) Dissolve H1 in dichloromethane, add 4-dimethylaminopyridine and tert-butyl dimethyl chlorosilane, stir at room temperature, purify the product after the reaction is completed, and obtain intermediate product H2; (3) Dissolve H2 in dichloromethane, add freshly prepared pyridine chlorochromate, stir at room temperature, purify the product after the reaction is completed, and obtain intermediate product H3; (4) Dissolve H3 in acetone, then add hydrochloric acid, stir at room temperature, purify the product after the reaction is completed, and obtain intermediate product H4; (5) Dissolve H4 in dichloromethane, and add dimethylaminopyridine, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and reactants in sequence. Stir at room temperature. After the reaction is completed, dilute, wash and dry the product in sequence to obtain crude product. The reactants are 1H-pyrazole-4-carboxylic acid, thiazole-4-carboxylic acid, oxazole-4-carboxylic acid, furan-3-carboxylic acid, pyrimidine-2-carboxylic acid, pyrazine-2-carboxylic acid or indole-2-carboxylic acid. (6) The crude product was purified by silica gel column chromatography to obtain the target compound.

6. The use of the hederone aglycone C-23 heterocyclic ester derivative of claim 2 in the preparation of P-gp inhibitors, wherein the hederone aglycone C-23 heterocyclic ester derivative is: 3-Carbonyl-23-((1H-pyrazole-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((thiazolyl-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((oxazol-4-carbonyl)oxy)-oleanolic-12-ene-28-carboxylic acid benzyl ester; 3-Carbonyl-23-((pyrimidin-2-carbonyl)oxy)-oleanolic-12-en-28-benzyl ester; Alternatively, 3-carbonyl-23-((pyrazin-2-carbonyl)oxy)-oleanolic-12-ene-28-benzyl ester.

Images