A derivative of a hedera saponin and uses thereof

Abstract

The application provides a hedera saponin derivative and application thereof, and relates to the field of medicinal chemistry. Different amino acid substances are used as connecting chains to modify (R)-(+) 1,2-dithiolane-3-pentanoic acid on the C-23 position of hedera saponin, and the hydrogen on the C-28 position carboxyl is substituted by a benzyl group, and the hydroxyl on the C-3 position is oxidized into a carbonyl group, so as to obtain a hedera saponin derivative. The hedera saponin derivative prepared by the application has high anti-inflammatory activity, can inhibit the inflammatory reaction caused by lipopolysaccharide at a lower concentration, has no biological toxicity to cells, and has a wide application prospect in the field of anti-inflammatory drugs.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and more particularly to an ivy saponin derivative and its applications. Background Technology

[0002] Inflammation is a defensive process that activates the physiological and pathological immune system. It is a defensive response of the body to external stimuli, characterized by high regulation and self-limitation, and generally has a protective function. However, excessive inflammatory responses or prolonged inflammation can lead to various diseases, such as immune dysfunction, sepsis, organ failure, and even death. Currently, the main drugs for treating inflammation are nonsteroidal anti-inflammatory drugs (NSAIDs), but long-term use or overdose can induce numerous side effects such as hypertension and arteriosclerosis. Therefore, there is an urgent need to develop anti-inflammatory drugs with novel structures, good efficacy, and fewer side effects.

[0003] Lipopolysaccharide (LPS) is a unique chemical component found in the exocellular layer of Gram-negative bacteria. When bacteria die, it detaches through cell lysis and destruction, exerting its toxicity by acting on animal cells, among other things. LPS possesses both thermal and chemical stability; it cannot be inactivated by conventional autoclaving or dry heat sterilization. Inactivation requires heating at 250°C for 30 minutes. The TLR family is associated with the expression of inflammatory cytokines and plays a crucial role in innate immunity. Detached LPS induces an inflammatory response in organisms via TLR4 (Toll-like receptor 4), which is present in the cell membrane of target cells.

[0004] Hederagenin (HG) is derived from *Hedera helix*, a plant belonging to the genus *Hedera* in the family Araliaceae. It is an oleanane-type pentacyclic triterpenoid compound. Previous studies have found that HG derivatives possess certain anti-inflammatory activities, and the mechanism of action has been preliminarily elucidated (see Yu T, Cheng HR, Li XL, et al. Design and synthesis of hederagenin derivatives modulating STING / NF-κB signaling for the relief of acute liver injury in septic mice[J]. European Journal of Medicinal Chemistry, 2023, 245, 114911). Therefore, it is of great significance to disclose a method for preparing hederagenin derivatives, expand the research on the anti-inflammatory activity of HG derivatives, and explore their applications in the anti-inflammatory field. Summary of the Invention

[0005] The purpose of this invention is to provide an ivy saponin derivative and its application, to improve the anti-inflammatory activity of existing ivy saponin compounds, and to broaden their application in the preparation of anti-inflammatory drugs, so as to solve the problem that non-steroidal anti-inflammatory drugs may induce side effects such as hypertension and arteriosclerosis when used for a long time or abused in excessive amounts.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides an ivy saponin derivative, the general formula of which is shown in Formula I:

[0008]

[0009] R1 includes One of them;

[0010] R2 is

[0011] This invention also provides the application of the above-mentioned ivy saponin derivatives in the preparation of anti-inflammatory drugs or pharmaceutically acceptable carriers.

[0012] The present invention has at least the following beneficial effects:

[0013] The ivy saponin derivatives disclosed in this invention have strong anti-inflammatory activity. Lipopolysaccharide in mouse mononuclear macrophages can induce cellular inflammation, thereby producing the inflammatory factor NO. When the ivy saponin derivatives disclosed in this invention are added to the culture medium and then stimulated with LPS, the NO level is lower than that in the experimental group without ivy saponin derivatives, confirming that the ivy saponin derivatives disclosed in this invention have anti-inflammatory activity. In addition, the cytotoxicity of the above-mentioned ivy saponin derivatives was also tested, and they showed good safety. Attached Figure Description

[0014] Figure 1 The graph shows the toxicity test results of the ivy saponin derivatives prepared in Examples 1-7 of this invention on RAW264.7 cells;

[0015] Figure 2 The graph shows the test results of the hederaponin derivatives prepared in Examples 1-7 of this invention inhibiting LPS-induced inflammation.

[0016] Figure 3 The graph shows the effect of different concentrations of the ivy saponin derivatives prepared in Example 3 on NO production in cells. Detailed Implementation

[0017] This invention provides an ivy saponin derivative, the general formula of which is shown in Formula I:

[0018]

[0019] R1 includes One of them;

[0020] R2 is

[0021] The preparation method of the ivy saponin derivative is as follows:

[0022] 1) Hederogenin, inorganic base, benzyl bromide and organic solvent are mixed and reacted to obtain 28-hederogenin benzyl ester;

[0023] 2) 28-hederone benzyl ester, 4-dimethylaminopyridine, tert-butyldimethylchlorosilane were mixed with an organic solvent and reacted to obtain 23-((tert-butyldimethylsilyl)oxy)-28-hederone benzyl ester;

[0024] 3) 23-((tert-butyldimethylsilyl)oxy)-28-hederone benzyl ester, pyridine chlorochromate, and organic solvent were mixed and oxidized to obtain 3-carbonyl-23-((tert-butyldimethylsilyl)oxy)-28-hederone benzyl ester;

[0025] 4) Mix 3-carbonyl-23-((tert-butyldimethylsilyl)oxy)-28-hederone benzyl ester, acid solution and organic solvent, and react to remove tert-butyldimethylsilyl group to obtain 3-carbonyl-28-hederone benzyl ester;

[0026] 5) 3-carbonyl-28-hederonide benzyl ester, catalyst, amino acid compound and organic solvent are mixed and reacted to obtain intermediate product 1, wherein the amino acid compound is one of BOC-glycine, BOC-β-alanine, BOC-4-aminobutyric acid, BOC-5-aminovaleric acid, N-Boc-4-aminobenzoic acid, BOC-glycylglycine and BOC-L-isoleucine;

[0027] 6) Mix the intermediate product 1 obtained in step 5) with the catalyst and organic solvent, react, remove the tert-butyloxycarbonyl protecting group, and obtain intermediate product 2;

[0028] 7) The intermediate product 2, catalyst, (R)-(+)-1,2-dithiopentane-3-pentanoic acid and organic solvent were reacted to obtain the ivy saponin derivative.

[0029] In this invention, the inorganic base in step 1) is sodium carbonate.

[0030] In this invention, the organic solvent in step 1) is N,N-dimethylimide.

[0031] In this invention, the molar volume ratio of ivy saponin, inorganic base, benzyl bromide and organic solvent in step 1) is 0.8-1.5 mmol: 1-3 mmol: 1-1.8 mmol: 12-15 mL.

[0032] In this invention, the reaction temperature in step 1) is 45-55°C and the reaction time is 6-10 hours.

[0033] In this invention, after the reaction in step 1) is completed, the mixture is further purified. Specifically, ethyl acetate is added to the mixture for dilution, the mixture is washed with water until the aqueous layer is neutral, and then washed with saturated brine to retain the organic layer. Anhydrous sodium sulfate is added to the organic layer for drying, followed by filtration, evaporation and concentration, column chromatography purification, solvent evaporation and other operations to obtain the purified product.

[0034] In the above purification process, the chromatography solvent used in the column chromatography purification process is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 5 to 10:1.

[0035] In this invention, the molar volume ratio of 28-hederone benzyl ester, 4-dimethylaminopyridine, tert-butyldimethylchlorosilane and organic solvent in step 2) is 0.6-1.0 mmol: 0.8-1.2 mmol: 2.0-3.0 mmol: 15-25 mL.

[0036] In this invention, the reaction temperature in step 2) is 20-30°C, and the reaction time is 4-8 hours.

[0037] In this invention, after the reaction in step 2) is completed, the mixture is further purified. Specifically, ethyl acetate is added to the mixture for dilution, acid washing is performed until acidic, and then saturated brine is used to wash and retain the organic layer. Anhydrous sodium sulfate is added to the organic layer for drying, followed by filtration, evaporation and concentration, column chromatography purification, solvent evaporation and other operations to obtain the purified product.

[0038] In the above purification process, the acid washing is performed using a 5 wt% hydrochloric acid solution.

[0039] In the above purification process, the chromatographic solvent used in the column chromatography purification is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 15 to 30:1.

[0040] In this invention, the molar volume ratio of 23-((tert-butyldimethylsilyl)oxy)-28-hederone benzyl ester, pyridine chlorochromate, and organic solvent in step 3) is 0.4–0.8 mmol: 1.0–1.6 mmol: 12–18 mL.

[0041] In this invention, the oxidation reaction temperature in step 3) is 20-30°C, and the oxidation reaction time is 6-10 hours.

[0042] In this invention, after the reaction in step 3) is completed, the mixture is further purified. Specifically, ethyl acetate is added to the mixture for dilution, the mixture is washed with water until the aqueous layer is neutral, and then washed with saturated brine to retain the organic layer. Anhydrous sodium sulfate is added to the organic layer for drying, followed by filtration, evaporation and concentration, column chromatography purification, solvent evaporation and other operations to obtain the purified product.

[0043] In the above purification process, the chromatographic solvent used in the column chromatography purification is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 20 to 35:1.

[0044] In this invention, the acid solution in step 4) is a hydrochloric acid solution, and the concentration of the acid solution is 8-15 wt%.

[0045] In this invention, the molar volume ratio of 3-carbonyl-23-((tert-butyldimethylsilyl)oxy)-28-hederone benzyl ester, acid solution and organic solvent in step 4) is 0.02-0.06 mmol: 1-3 mL: 8-15 mL.

[0046] In this invention, the reaction temperature in step 4) is 20-30°C, and the reaction time is 3-5 hours.

[0047] In this invention, after the reaction in step 4) is completed, the mixture is further purified. Specifically, ethyl acetate is added to the mixture for dilution, the mixture is washed with water until the aqueous layer is neutral, and then saturated brine is used to wash the organic layer. Anhydrous sodium sulfate is added to the organic layer for drying, followed by filtration, evaporation and concentration, column chromatography purification, solvent evaporation and other operations to obtain the purified product.

[0048] In the above purification process, the chromatographic solvent used in the column chromatography purification is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 5 to 8:1.

[0049] In this invention, the catalyst in step 5) is a mixed catalyst of 4-dimethylaminopyridine and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride;

[0050] The molar ratio of 4-dimethylaminopyridine to 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 0.8–1.2:0.8–1.2.

[0051] In this invention, the molar volume ratio of 3-carbonyl-28-hederone benzyl ester, catalyst, amino acid compound and organic solvent in step 5) is 0.3-0.8 mmol: 1.6-2.4 mmol: 0.6-1.5 mmol.

[0052] In this invention, the reaction temperature in step 5) is 20-30°C, and the reaction time is 3-5 hours.

[0053] In this invention, after the reaction in step 5) is completed, the mixture is further purified. Specifically, ethyl acetate is added to the mixture for dilution, the mixture is washed with water until the aqueous layer is neutral, and then saturated brine is used to wash the organic layer. Anhydrous sodium sulfate is added to the organic layer for drying, followed by filtration, evaporation and concentration, column chromatography purification, solvent evaporation and other operations to obtain the purified product.

[0054] In the above purification process, the chromatography solvent used in the column chromatography purification process is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 10 to 15:1.

[0055] In this invention, the catalyst in step 6) is trifluoroacetic acid.

[0056] In this invention, the molar volume ratio of intermediate product 1, catalyst and organic solvent in step 6) is 0.3-0.7 mmol: 0.5-1.5 mL: 7-14 mL.

[0057] In this invention, the reaction temperature in step 6) is 20-30°C, and the reaction time is 2-3 hours.

[0058] In this invention, after the reaction in step 6) is completed, the mixture is further purified. Specifically, ethyl acetate is added to the mixture for dilution, the mixture is washed with water until the aqueous layer is neutral, and then washed with saturated brine to retain the organic layer. Anhydrous sodium sulfate is added to the organic layer for drying, and then the mixture is filtered, concentrated by evaporation, purified by column chromatography, and the solvent is evaporated to obtain the purified product.

[0059] In the above purification process, the chromatography solvent used in the column chromatography purification process is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 8 to 10:1.

[0060] In this invention, the catalyst in step 7) is a mixed catalyst of 4-dimethylaminopyridine and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride;

[0061] The molar ratio of 4-dimethylaminopyridine to 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 0.6–1.0:0.6–1.0.

[0062] In this invention, the molar volume ratio of intermediate product 2, catalyst, (R)-(+)-1,2-dithiopentane-3-pentanoic acid and organic solvent in step 7) is 0.3-0.5 mmol: 1.2-2.0 mmol: 0.6-1.2 mmol: 5-12 mL.

[0063] In this invention, the reaction temperature in step 7) is 20-30°C, and the reaction time is 3-5 hours.

[0064] In this invention, the organic solvents in steps 2) to 7) are independently selected from one or more of dichloromethane and acetone.

[0065] In this invention, after the reaction in step 7) is completed, the mixture is further purified. Specifically, ethyl acetate is added to the mixture for dilution, the mixture is washed with water until the aqueous layer is neutral, and then washed with saturated brine to retain the organic layer. Anhydrous sodium sulfate is added to the organic layer for drying, followed by filtration, evaporation and concentration, column chromatography purification, solvent evaporation and other operations to obtain the purified product.

[0066] In the above purification process, the chromatography solvent used in the column chromatography purification process is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 10 to 15:1.

[0067] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0068] Example 1

[0069] 3-Carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)glycyl)oxy)-28-hederonium benzyl ester, prepared by the following method:

[0070] 1) Dissolve 1.0 mmol of hederaponin in 15.0 mL of N,N-dimethylformamide, add 2.1 mmol of potassium carbonate and 1.3 mmol of benzyl bromide, and stir at 50 °C for 8 h. After the reaction is complete, dilute the reaction solution with 25.0 mL of ethyl acetate, wash three times with water and twice with saturated brine, dry the organic layer with anhydrous sodium sulfate, filter, remove the solvent under reduced pressure, and purify the product by silica gel column chromatography (using a 10:1 volume ratio of petroleum ether and ethyl acetate as the chromatographic solvent). Heat the receiving solution containing the target substance to evaporate the solvent, and obtain 470.0 mg of white solid 28-hederaponin benzyl ester (yield 83.38%).

[0071] 2) Dissolve 0.8 mmol of 28-hederonide benzyl ester prepared in step 1) in 20.0 mL of dichloromethane, add 1.0 mmol of 4-dimethylaminopyridine and 2.4 mmol of tert-butyldimethylchlorosilane, and stir at room temperature (25 °C) for 6 h. After the reaction is complete, evaporate the dichloromethane, add 20 mL of ethyl acetate to dilute, and wash three times with 5% HCl until the aqueous layer is acidic. Wash the organic layer with saturated brine until the aqueous layer is neutral. Separate the layers to obtain the organic layer, add anhydrous sodium sulfate to the organic layer for drying, filter, concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a mixture of petroleum ether and ethyl acetate with a volume ratio of 25:1). Heat the receiving liquid containing the target substance to evaporate the solvent, and obtain 383.0 mg of white solid 23-((tert-butyldimethylsilyl)oxy)-28-hederonide benzyl ester (yield 70.72%).

[0072] 3) Dissolve 0.6 mmol of 23-((tert-butyldimethylsilyl)oxy)-28-hederonium benzyl ester prepared in step 2) above in 15.0 mL of dichloromethane, add 1.3 mmol of freshly prepared pyridine chlorochromate, and stir at room temperature (25 °C) for 8 h. After the reaction is complete, remove the dichloromethane by evaporation, dilute with 20 mL of ethyl acetate, wash 3 times with water, and then wash 2 times with saturated brine until neutral. Separate the liquid to obtain the organic layer, add anhydrous sodium sulfate to the organic layer and dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a mixture of petroleum ether and ethyl acetate with a volume ratio of 30:1). Heat the receiving liquid containing the target substance to evaporate and remove the solvent, to obtain 319.0 mg of white solid 3-carbonyl-23-((tert-butyldimethylsilyl)oxy)-28-hederonium benzyl ester (yield 78.77%).

[0073] 4) Dissolve 0.4 mmol of 3-carbonyl-23-((tert-butyldimethylsilyl)oxy)-28-hederonium benzyl ester prepared in step 3) above in 10.0 mL of acetone, add 2.0 mL of 10 wt% HCl solution, and stir at room temperature (25 °C) for 4 h. After the reaction is complete, evaporate the solvent, add 20 mL of ethyl acetate to dilute, wash with water 3 times until the lower aqueous solution is neutral, then wash with saturated brine 2 times, separate to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a 5:1 volume ratio of petroleum ether and ethyl acetate). Heat the receiving liquid containing the target substance to evaporate the solvent, and obtain 225.0 mg of white solid 3-carbonyl-28-hederonium benzyl ester (yield 89.0%).

[0074] 5) Dissolve 0.5 mmol of 3-carbonyl-28-hederone benzyl ester prepared in step 4) in 8.0 mL of dichloromethane, add 1 mmol DMAP (4-dimethylaminopyridine), 1 mmol EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide), and 0.8 mmol Boc-glycine, and stir at room temperature (23 °C) for 4 h. After the reaction was completed, the solvent was evaporated, 20 mL of ethyl acetate was added for dilution, and the mixture was washed three times with water until the lower aqueous solution was neutral. Then, it was washed three times with saturated saline solution. The organic layer was separated, and anhydrous sodium sulfate was added to the organic layer for drying. After filtration, evaporation and concentration, the product was purified by silica gel column chromatography (the chromatographic solvent was a mixture of petroleum ether and ethyl acetate at a volume ratio of 10:1). The receiving solution containing the target substance was heated and evaporated to remove the solvent, yielding 330.0 mg of white solid 3-carbonyl-23-(((tert-butoxycarbonyl)glycyl)oxy)-28-hederonium benzyl glycine ester (yield 92.0%).

[0075] 6) Dissolve 0.5 mmol of 3-carbonyl-23-(((tert-butoxycarbonyl)glycyl)oxy)-28-hederogen benzyl ester glycine ester prepared in step 5) above in 10.0 mL of dichloromethane, add 1.0 mL of trifluoroacetic acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, add 20 mL of ethyl acetate to dilute, wash with water 4 times until the lower aqueous solution is neutral, wash with saturated brine 2 times, separate to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a 10:1 volume ratio of petroleum ether and ethyl acetate). Heat the receiving liquid containing the target substance to evaporate the solvent, and obtain 280.0 mg of white solid 3-carbonyl-23-glycyloxy-28-hederogen benzyl ester (yield 90.0%).

[0076] 7) Dissolve 0.4 mmol of 3-carbonyl-23-glycyloxy-28-hederone benzyl ester prepared in step 6) in 8.0 mL of dichloromethane, add 0.8 mmol DMAP, 0.8 mmol EDCI, and 0.8 mmol (R)-(+)-1,2-dithiopentane-3-pentanoic acid, and stir at room temperature (25 °C) for 5 h. After the reaction was completed, the solvent was evaporated, 20 mL of ethyl acetate was added for dilution, and the mixture was washed three times with water until the lower aqueous solution was neutral. Then, it was washed once with saturated brine. The organic layer was separated, and anhydrous sodium sulfate was added to the organic layer for drying. After filtration, evaporation and concentration, the product was purified by silica gel column chromatography (the chromatographic solvent was a mixture of petroleum ether and ethyl acetate at a volume ratio of 15:1). The receiving solution containing the target substance was heated and evaporated to remove the solvent, yielding 295.0 mg of a pale yellow solid {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)glycyl)oxy)-28-hederonium saponin benzyl ester} (yield 92.0%).

[0077] The above product {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)glycyl)oxy)-28-hederonium benzyl ester} was analyzed by 1H NMR spectroscopy, and the results are as follows:

[0078] 1 H NMR (400MHz, CDCl3) δ7.33(s,5H,H-Ar),6.05–6.01(m,1H,H-NH),5.32–5.29(m,1H,H-H12),5.11–5.02(m,2H,H-CH2Ar),4.20(d,J=1 0.7Hz,1H,H-H23a),4.06(d,J=10.7Hz,1H,H-H23b),4.05–3.91(m,2H,H-CH2N),3.56(s,1H,H-CHS),3.19–3.10(m,2H,H-CH2S),2.91 (dd,J=13.7,4.5Hz,1H,H-18),2.54–2.41(m,2H,H-1),2.36(t,J=7.4Hz,2H,CH2),2.24(t,J=7.5Hz,2H,CH2),1.92–1.89(m,2H,CH2) ,1.70–1.28(m,24H,CH2),1.13(s,3H,CH3),1.02(s,3H,CH3),1.01(s,3H,CH3),0.91(s,3H,CH3),0.89(s,3H,CH3),0.65(s,3H,CH3).

[0079] Example 2

[0080] 3-Carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)alanyl)oxy)-28-hederonium benzyl ester, prepared by the following method:

[0081] Steps 1) to 4) are the same as steps 1) to 4) in Example 1, to prepare 3-carbonyl-28-hederonide benzyl ester.

[0082] 5) Dissolve 0.65 mmol of 3-carbonyl-28-hederonide benzyl ester in 8.0 mL of dichloromethane, add 1 mmol DMAP, 1 mmol EDCI, and 0.8 mmol Boc-β-alanine, and stir at room temperature (24 °C) for 3 h. After the reaction is complete, evaporate the solvent, dilute with 20 mL of ethyl acetate, wash with water 4 times until the lower aqueous solution is neutral, wash once with saturated brine, separate to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a mixture of petroleum ether and ethyl acetate with a volume ratio of 8:1). Heat the receiving solution containing the target substance to evaporate the solvent, and obtain 430.5 mg of white solid 3-carbonyl-23-((3-((tert-butoxycarbonyl)amino)propionyl)oxy)-28-hederonide benzyl ester glycine ester (yield 90.6%).

[0083] 6) Dissolve 0.4 mmol of 3-carbonyl-23-((3-((tert-butoxycarbonyl)amino)propionyl)oxy)-28-hederogen benzyl ester glycine ester obtained in step 5) above in 10.0 mL of dichloromethane, add 1.0 mL of trifluoroacetic acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, add 20 mL of ethyl acetate to dilute, wash with water 3 times until the lower aqueous solution is neutral, wash with saturated brine 2 times, separate to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a 10:1 volume ratio of petroleum ether and ethyl acetate). Heat the receiving liquid containing the target substance to evaporate the solvent to obtain 220.0 mg of white solid 3-carbonyl-23-((3-aminopropionyl)oxy)-28-hederogen benzyl ester (yield 87.0%).

[0084] 7) Dissolve 0.6 mmol of 3-carbonyl-23-((3-aminopropionyl)oxy)-28-hederone benzyl ester prepared in step 6) above in 8.0 mL of dichloromethane, add 0.8 mmol DMAP, 0.8 mmol EDCI and 0.8 mmol (R)-(+)-1,2-dithiopentane-3-pentanoic acid and stir at room temperature (25 °C) for 4.5 h. After the reaction was completed, the solvent was evaporated, 20 mL of ethyl acetate was added for dilution, and the mixture was washed four times with water until the lower aqueous solution was neutral. Then, it was washed once with saturated brine. The organic layer was separated, and anhydrous sodium sulfate was added to the organic layer for drying. After filtration, evaporation and concentration, the product was purified by silica gel column chromatography (the chromatographic solvent was a mixture of petroleum ether and ethyl acetate at a volume ratio of 10:1). The receiving liquid containing the target substance was heated and evaporated to remove the solvent, yielding 450.0 mg of a pale yellow solid {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)alanyl)oxy)-28-hederonium saponin benzyl ester} (yield 91.6%).

[0085] The above product {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)alanyl)oxy)-28-hederonium benzyl ester} was analyzed by 1H NMR spectroscopy, and the results are as follows:

[0086] 1 H NMR (400MHz, CDCl3) δ7.33(s,5H,H-Ar),6.30(s,1H,H-NH),5.32–5.29(m,1H,H-12),5.11–5.02(m,2H,H-CH2Ar), 4.12–4.04(m,2H,H-23),3.57(s,1H,H-SCH),3.48(q,J=6.2Hz,2H,H-CH2N),3.11(s,2H,H-SCH2),2.91(dd,J=13. 8,4.5Hz,1H,H-18),2.51–2.43(m,4H,CH2),2.19(d,J=7.5Hz,2H,H-1),1.94–1.88(m,4H,CH2),1.71–1.26(m,24H ,CH2),1.13(s,3H,CH3),1.04(s,3H,CH3),1.01(s,3H,CH3),0.92(s,3H,CH3),0.90(s,3H,CH3),0.65(s,3H,CH3).

[0087] Example 3

[0088] 3-Carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)butanyl)oxy)-28-hederonium benzyl ester, prepared by the following method:

[0089] Steps 1) to 4) are the same as steps 1) to 4) in Example 1, to prepare 3-carbonyl-28-hederonide benzyl ester.

[0090] 5) Dissolve 0.65 mmol of 3-carbonyl-28-hederamine benzyl ester in 8.0 mL of dichloromethane, add 1 mmol DMAP, 1 mmol EDCI, and 0.8 mmol Boc-4-aminobutyric acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, dilute with 20 mL of ethyl acetate, wash with water 4 times until the lower aqueous solution is neutral, wash twice with saturated brine, separate the liquid to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a mixture of petroleum ether and ethyl acetate with a volume ratio of 8:1). Heat the receiving liquid containing the target substance to evaporate the solvent, and obtain 420.0 mg of white solid 3-carbonyl-23-((4-((tert-butoxycarbonyl)amino)butyryl)oxy)-28-hederamine benzyl ester butyric acid ester (yield 86.7%).

[0091] 6) Dissolve 0.4 mmol of 3-carbonyl-23-((4-((tert-butoxycarbonyl)amino)butyryl)oxy)-28-hederogen benzyl ester butyric acid ester prepared in step 5) in 10.0 mL of dichloromethane, add 1.0 mL of trifluoroacetic acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, add 20 mL of ethyl acetate to dilute, wash three times with water until the lower aqueous solution is neutral, wash twice with saturated brine, separate the liquid to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a 10:1 volume ratio of petroleum ether and ethyl acetate). Heat the receiving liquid containing the target substance to evaporate the solvent, and obtain 220.0 mg of white solid 3-carbonyl-23-((4-aminobutyryl)oxy)-28-hederogen benzyl ester (yield 85.0%).

[0092] 7) Dissolve 0.6 mmol of 3-carbonyl-23-((4-aminobutyryl)oxy)-28-hederone benzyl ester prepared in step 6) above in 8.0 mL of dichloromethane, add 0.8 mmol DMAP, 0.8 mmol EDCI and 0.8 mmol (R)-(+)-1,2-dithiopentane-3-pentanoic acid and stir at room temperature (25 °C) for 4.5 h. After the reaction was completed, the solvent was evaporated, 20 mL of ethyl acetate was added for dilution, and the mixture was washed four times with water until the lower aqueous solution was neutral. Then, it was washed once with saturated brine. The organic layer was separated, and anhydrous sodium sulfate was added to the organic layer for drying. After filtration, evaporation and concentration, the product was purified by silica gel column chromatography (the chromatographic solvent was a mixture of petroleum ether and ethyl acetate at a volume ratio of 10:1). The receiving liquid containing the target substance was heated and evaporated to remove the solvent, yielding 450.0 mg of a pale yellow solid {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)butanyl)oxy)-28-hederonium saponin benzyl ester} (yield 91.6%).

[0093] The above product {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)butanyl)oxy)-28-hederonium benzyl ester} was analyzed by 1H NMR spectroscopy, and the results are as follows:

[0094] 1 H NMR (400MHz, CDCl3) δ7.34(s,5H,H-Ar),5.82(d,J=5.8Hz,1H,H-NH),5.31(s,1H,H-12),5.12–5.03(m,2H,H-CH 2Ar),4.06(s,2H,H-23),3.62–3.51(m,1H,H-SCH),3.32–3.22(m,2H,H-SCH2),3.12(s,2H,H-CH2N),2.95–2.89( m,1H,H-18),2.53–2.40(m,2H,H-1),2.31(d,J=7.1Hz,2H,CH2),2.17(d,J=7.5Hz,2H,CH2),1.95–1.34(m,30H,C H2),1.25(s,3H,CH3),1.12(s,3H,CH3),1.02(s,3H,CH3),0.92(s,3H,CH3),0.90(s,3H,CH3),0.65(s,3H,CH3).

[0095] Example 4

[0096] 3-Carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)pentanoyl)oxy)-28-hederonium benzyl ester, prepared by the following method:

[0097] Steps 1) to 4) are the same as steps 1) to 4) in Example 1, to prepare 3-carbonyl-28-hederonide benzyl ester.

[0098] 5) Dissolve 0.65 mmol of 3-carbonyl-28-hederonide benzyl ester in 8.0 mL of dichloromethane, add 1 mmol DMAP, 1 mmol EDCI, and 0.8 mmol Boc-4-aminovaleric acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, dilute with 20 mL of ethyl acetate, wash with water 4 times until the lower aqueous solution is neutral, wash twice with saturated brine, separate the liquid to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a mixture of petroleum ether and ethyl acetate with a volume ratio of 8:1). Heat the receiving liquid containing the target substance to evaporate the solvent, and obtain 460.0 mg of white solid 3-carbonyl-23-((5-((tert-butoxycarbonyl)amino)valeryl)oxy)-28-hederonide benzyl ester pentanoic acid ester (yield 93.3%).

[0099] 6) Dissolve 0.4 mmol of 3-carbonyl-23-((5-((tert-butoxycarbonyl)amino)valerate)oxy)-28-hederogen benzyl ester pentanoic acid ester prepared in step 5) above in 10.0 mL of dichloromethane, add 1.0 mL of trifluoroacetic acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, add 20 mL of ethyl acetate to dilute, wash with water 3 times until the lower aqueous solution is neutral, then wash with saturated brine 2 times, separate to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a 10:1 volume ratio of petroleum ether and ethyl acetate). Heat the receiving liquid containing the target substance to evaporate the solvent to obtain 240.0 mg of white solid 3-carbonyl-23-((4-aminovalerate)oxy)-28-hederogen benzyl ester (yield 91.2%).

[0100] 7) Dissolve 0.6 mmol of 3-carbonyl-23-((4-aminopentanoyl)oxy)-28-hederone benzyl ester prepared in step 6) above in 8.0 mL of dichloromethane, add 0.8 mmol DMAP, 0.8 mmol EDCI and 0.8 mmol (R)-(+)-1,2-dithiopentane-3-pentanoic acid and stir at room temperature (25 °C) for 4.5 h. After the reaction was completed, the solvent was evaporated, 20 mL of ethyl acetate was added for dilution, and the mixture was washed three times with water until the lower aqueous solution was neutral. Then, it was washed twice with saturated brine. The organic layer was separated, and anhydrous sodium sulfate was added to the organic layer for drying. After filtration, evaporation and concentration, the product was purified by silica gel column chromatography (the chromatographic solvent was a mixture of petroleum ether and ethyl acetate at a volume ratio of 10:1). The receiving liquid containing the target substance was heated and evaporated to remove the solvent, yielding 470.0 mg of a pale yellow solid {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)pentanoyl)oxy)-28-hederonium saponin benzyl ester} (yield 92.5%).

[0101] The above product {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)butanyl)oxy)-28-hederonium benzyl ester} was analyzed by 1H NMR spectroscopy, and the results are as follows:

[0102] 1 H NMR (400MHz, CDCl3) δ7.34(s,5H,H-Ar),5.70(s,1H,H-NH),5.32(t,J=3.6Hz,1H,H-H12),5.14–5.02(m,2H,H-CH2Ar),4.07(d, J=3.9Hz,2H,H-H23),3.57(dd,J=8.2,6.2Hz,1H,H-CHS),3.24(d,J=6.2Hz,2H,H-CH2N),3.20–3.07(m,2H,H-CH2S),2.92(dd,J =13.9,4.5Hz,1H,H-18),2.45(dd,J=12.8,6.3Hz,2H,H-1),2.31(t,J=7.0Hz,2H,CH2),2.18(t,J=7.5Hz,2H,CH2),1.98–1.88( m,4H,CH2),1.71–1.25(m,28H,CH2),1.13(s,3H,CH3),1.02(s,6H,CH3),0.92(s,3H,CH3),0.90(s,3H,CH3),0.66(s,3H,CH3).

[0103] Example 5

[0104] 3-Carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)hexanoyl)oxy)-28-hederonium benzyl ester, prepared by the following method:

[0105] Steps 1) to 4) are the same as steps 1) to 4) in Example 1, to prepare 3-carbonyl-28-hederonide benzyl ester.

[0106] 5) Dissolve 0.65 mmol of 3-carbonyl-28-hederonium benzyl ester in 8.0 mL of dichloromethane, add 1 mmol DMAP, 1 mmol EDCI, and 0.8 mmol Boc-6-aminohexanoic acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, dilute with 20 mL of ethyl acetate, wash three times with water until the lower aqueous solution is neutral, wash once with saturated brine, separate to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a 10:1 volume ratio of petroleum ether and ethyl acetate). Heat the receiving solution containing the target substance to evaporate the solvent, and obtain 460.0 mg of white solid 3-carbonyl-23-((6-((tert-butoxycarbonyl)amino)hexanoyl)oxy)-28-hederonium benzyl ester hexanoic acid ester (yield 91.6%).

[0107] 6) Dissolve 0.4 mmol of 3-carbonyl-23-((6-((tert-butoxycarbonyl)amino)hexanoyl)oxy)-28-hederonium benzyl ester hexanoic acid ester prepared in step 5) above in 10.0 mL of dichloromethane, add 1.0 mL of trifluoroacetic acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, add 20 mL of ethyl acetate to dilute, wash with water 3 times until the lower aqueous solution is neutral, then wash with saturated brine 2 times, separate to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a 10:1 volume ratio of petroleum ether and ethyl acetate). Heat the receiving liquid containing the target substance to evaporate the solvent to obtain 235.0 mg of white solid 3-carbonyl-23-((4-aminopentanoyl)oxy)-28-hederonium benzyl ester (yield 87.3%).

[0108] 7) Dissolve 0.6 mmol of 3-carbonyl-23-((4-aminopentanoyl)oxy)-28-hederone benzyl ester prepared in step 6) above in 8.0 mL of dichloromethane, add 0.8 mmol DMAP, 0.8 mmol EDCI and 0.8 mmol (R)-(+)-1,2-dithiopentane-3-pentanoic acid and stir at room temperature (25 °C) for 4.5 h. After the reaction was completed, the solvent was evaporated, 20 mL of ethyl acetate was added for dilution, and the mixture was washed three times with water until the lower aqueous solution was neutral. Then, it was washed twice with saturated brine. The organic layer was separated, and anhydrous sodium sulfate was added to the organic layer for drying. After filtration, evaporation and concentration, the product was purified by silica gel column chromatography (the chromatographic solvent was a mixture of petroleum ether and ethyl acetate at a volume ratio of 10:1). The receiving liquid containing the target substance was heated and evaporated to remove the solvent, yielding 476.0 mg of a pale yellow solid {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)hexanoyl)oxy)-28-hederonium saponin benzyl ester} (yield 92.2%).

[0109] The above product {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)hexanoyl)oxy)-28-hederonium benzyl ester} was analyzed by 1H NMR spectroscopy, and the results are as follows:

[0110] 1 H NMR (400MHz, CDCl3) δ7.34(s,5H,H-Ar),5.76(s,1H,H-NH),5.32(t,J=3.6Hz,1H,H-H12),5.12–5.03(m,2H,H-CH2Ar),4. 06(s,2H,H-H23),3.56(dd,J=8.3,6.3Hz,1H,H-SCH),3.28–3.19(m,2H,H-SCH2),3.18–3.07(m,2H,H-CH2N),2.95–2.89( m,1H,H-18),2.50–2.40(m,2H,H-1),2.28(t,J=7.2Hz,2H,CH2),2.17(t,J=7.3Hz,2H,CH2),1.97–1.88(m,4H,CH2),1.72 –1.24(m,30H,CH2),1.12(s,3H,CH3),1.01(d,J=2.0Hz,6H,CH3×2),0.92(s,3H,CH3),0.90(s,3H,CH3),0.66(s,3H,CH3).

[0111] Example 6

[0112] 3-Carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)glycylglycyl)oxy)-28-hederonium benzyl ester, prepared by the following method:

[0113] Steps 1) to 4) are the same as steps 1) to 4) in Example 1, to prepare 3-carbonyl-28-hederonide benzyl ester.

[0114] 5) Dissolve 0.65 mmol of 3-carbonyl-28-hederamenoate benzyl ester in 8.0 mL of dichloromethane, add 1 mmol DMAP, 1 mmol EDCI, and 0.75 mmol Boc-6-aminohexanoic acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, dilute with 20 mL of ethyl acetate, wash three times with water until the lower aqueous solution is neutral, wash once with saturated brine, separate the liquid to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a 10:1 volume ratio of petroleum ether and ethyl acetate). Heat the receiving liquid containing the target substance to evaporate the solvent, and obtain 475.0 mg of white solid 3-carbonyl-23-((6-((tert-butoxycarbonyl)glycylglycyl)oxy)-28-hederamenoate benzyl hexanoate ester (yield 94.4%).

[0115] 6) Dissolve 0.4 mmol of 3-carbonyl-23-((6-((tert-butoxycarbonyl)glycylglycyl)oxy)-28-hederogen benzyl ester hexanoic acid ester prepared in step 5) above in 10.0 mL of dichloromethane, add 1.0 mL of trifluoroacetic acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, add 20 mL of ethyl acetate to dilute, wash with water 3 times until the lower aqueous solution is neutral, wash with saturated brine 2 times, separate the liquid to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a 10:1 volume ratio of petroleum ether and ethyl acetate). Heat the receiving liquid containing the target substance to evaporate the solvent to obtain 235.0 mg of white solid 3-carbonyl-23-((glycylglycyl)oxy)-28-hederogen benzyl ester (yield 87.0%).

[0116] 7) Dissolve 0.6 mmol of 3-carbonyl-23-((glycylglycyl)oxy)-28-hederone benzyl ester prepared in step 6) above in 8.0 mL of dichloromethane, add 0.8 mmol DMAP, 0.8 mmol EDCI, and 0.8 mmol (R)-(+)-1,2-dithiopentane-3-pentanoic acid, and stir at room temperature (25 °C) for 4.5 h. After the reaction was completed, the solvent was evaporated, 20 mL of ethyl acetate was added for dilution, and the mixture was washed three times with water until the lower aqueous solution was neutral. Then, it was washed twice with saturated brine. The organic layer was separated, and anhydrous sodium sulfate was added to the organic layer for drying. After filtration, evaporation and concentration, the product was purified by silica gel column chromatography (the chromatographic solvent was a mixture of petroleum ether and ethyl acetate at a volume ratio of 10:1). The receiving solution containing the target substance was heated and evaporated to remove the solvent, yielding 470.0 mg of a pale yellow solid {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)glycylglycyl)oxy)-28-hederonium saponin benzyl ester} (yield 90.9%).

[0117] The above product {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)glycylglycyl)oxy)-28-hederonium benzyl ester} was analyzed by 1H NMR spectroscopy, and the results are as follows:

[0118] 1 H NMR (400MHz, CDCl3) δ7.33(s,5H,H-Ar),6.75(t,J=5.4Hz,1H,H-NHa),6.46(t,J=5.3Hz,1H,H-NHb),5.32–5.29(m,1H,H-12),5.12–5.02(m, 2H,H-CH2Ar),4.21–4.05(m,2H,H-23),4.03–3.98(m,2H,H-CH2Na),3.98–3.93(m,2H,H-CH2Nb),3.60–3.53(m,1H,H-SCH),3.18–3.08(m,2H, H-SCH2),2.91(dd,J=13.6,4.4Hz,1H,H-18),2.54–2.41(m,2H,H-1),2.36(t,J=7.4Hz,2H,CH2),2.27(t,J=7.5Hz,2H,CH2),1.94–1.86(m,4 H,CH2),1.70–1.21(m,24H,CH2),1.14(s,3H,CH3),1.02(s,3H,CH3),1.01(s,3H,CH3),0.92(s,3H,CH3),0.90(s,3H,CH3),0.65(s,3H,CH3).

[0119] Example 7

[0120] 3-Carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)isoleucyl)oxy)-28-hederonium benzyl ester, prepared by the following method:

[0121] Steps 1) to 4) are the same as steps 1) to 4) in Example 1, to prepare 3-carbonyl-28-hederonide benzyl ester.

[0122] 5) Dissolve 0.65 mmol of 3-carbonyl-28-hederonium benzyl ester in 8.0 mL of dichloromethane, add 1 mmol DMAP, 1 mmol EDCI, and 0.75 mmol Boc-L-isoleucine, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, dilute with 20 mL of ethyl acetate, wash 5 times with water until the lower aqueous solution is neutral, wash once with saturated brine, separate the liquid to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a 10:1 volume ratio of petroleum ether and ethyl acetate). Heat the receiving liquid containing the target substance to evaporate the solvent, and obtain 480.0 mg of white solid 3-carbonyl-23-((6-((tert-butoxycarbonyl)-L-isoleucyl)oxy)-28-hederonium benzyl ester hexanoic acid ester (yield 95.6%).

[0123] 6) Dissolve 0.4 mmol of 3-carbonyl-23-((6-((tert-butoxycarbonyl)-L-isoleucyl)oxy)-28-hederogen benzyl ester hexanoic acid ester prepared in step 5) above in 10.0 mL of dichloromethane, add 1.0 mL of trifluoroacetic acid, and stir at room temperature (25 °C) for 3 h. After the reaction is complete, evaporate the solvent, add 20 mL of ethyl acetate to dilute, wash with water 3 times until the lower aqueous solution is neutral, wash with saturated brine 2 times, separate the liquid to obtain the organic layer, add anhydrous sodium sulfate to the organic layer to dry, filter, evaporate and concentrate, and purify the product by silica gel column chromatography (the chromatographic solvent is a mixture of petroleum ether and ethyl acetate with a volume ratio of 10:1). Heat the receiving liquid containing the target substance to evaporate the solvent to obtain 235.0 mg of white solid 3-carbonyl-23-((L-isoleucyl))oxy)-28-hederogen benzyl ester (yield 87.0%).

[0124] 7) Dissolve 0.6 mmol of 3-carbonyl-23-((L-isoleucyl))oxy)-28-hederone benzyl ester prepared in step 6) above in 8.0 mL of dichloromethane, add 0.8 mmol DMAP, 0.8 mmol EDCI and 0.8 mmol (R)-(+)-1,2-dithiopentane-3-pentanoic acid and stir at room temperature (25 °C) for 4.5 h. After the reaction was completed, the solvent was evaporated, 20 mL of ethyl acetate was added for dilution, and the mixture was washed three times with water until the lower aqueous solution was neutral. Then, it was washed twice with saturated saline solution. The organic layer was separated, and anhydrous sodium sulfate was added to the organic layer for drying. After filtration, evaporation and concentration, the product was purified by silica gel column chromatography (the chromatographic solvent was a mixture of petroleum ether and ethyl acetate at a volume ratio of 10:1). The receiving solution containing the target substance was heated and evaporated to remove the solvent, yielding 480.0 mg of a pale yellow solid {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)isoleucyl)oxy)-28-hederonium saponin benzyl ester} (yield 93.0%).

[0125] The above product {3-carbonyl-23-(((5-(((R)-1,2-dithiocyclopentan-3-yl)pentanoyl)isoleucyl)oxy)-28-hederonium benzyl ester} was analyzed by 1H NMR spectroscopy, and the results are as follows:

[0126] 1H NMR (400MHz, CDCl3) δ7.34(s,5H,H-Ar),5.99(d,J=8.4Hz,1H,H-NH),5.31(d,J=3.6Hz,1H,H-12),5.12–5.03(m,2H,H-CH2Ar),4.58(dd,J=8.4,4.4Hz,1 H, CHN),4.33(d,J=10.7Hz,1H,H-23a),3.91(d,J=10.7Hz,1H,H-23b),3.61– 3.51(m,1H,H-SCH),3.21–3.07(m,2H,H-SCH2),2.92(dd,J=14.3,4.5Hz,1H, H-18),2.48(ddd,J=26.3,12.1,6.3Hz,2H,CH2),2.39–2.30(m,1H,H-1a),2. 23(t,J=7.4Hz,2H,CH2),2.01(dd,J=13.7,4.1Hz,1H,H-1b),1.92(d,J=6.2H z,3H,H-CH,H-CH2),1.72–1.25(m,26H,CH2),1.14(s,3H,CH3),1.05(s,3H,C H3),1.00(s,3H,CH3),0.92(s,3H,CH3),0.90(s,3H,CH3),0.66(s,3H,CH3).

[0127] The activities of several ivy saponin derivatives prepared in Examples 1-7 of this invention were tested. The specific test methods and results are as follows:

[0128] Experimental Example 1

[0129] The cytotoxicity of several hederone derivatives prepared in Examples 1-7 was tested, and the specific testing methods are as follows:

[0130] RAW264.7 cells (mouse macrophages) in the logarithmic growth phase were harvested, and the cell density was adjusted to 5 × 10⁶ cells per 100 μL. 4A suspension of cells was seeded into 96-well plates and incubated in a cell culture incubator (37°C, 5% CO2) for 24 h. Seven hedyotis diffusin derivatives prepared in Examples 1-7 were added to the cells, with a concentration of 40 μM / mL in the cell culture medium for each group. The cells were incubated in a cell culture incubator (37°C, 5% CO2) for 24 h. Each group was set up with 5 replicates. After 24 h, cell viability was assessed using the MTT assay. 20 μL of 5 mg / mL MTT solution was added to each well, and the cells were incubated in the incubator (37°C, 5% CO2) for another 4 h. The plate was removed from the incubator to terminate the reaction, and the culture medium was carefully aspirated from the wells. 150 μL of DMSO was added to each well, and the plates were shaken at low speed for 10 min to fully dissolve the crystals. The OD value was measured at 570 nm. The amount of MTT crystals formed was directly proportional to the number of viable cells. Cell viability was assessed as follows: Figure 1 As shown. In Examples 1-5 and Example 7, the cell viability of RAW264.7 cells at 40 μM was all above 90%. Compared with the blank control group, Examples 1-5 and Example 7 were safe and non-toxic to RAW264.7 cells at 40 μM.

[0131] Experiment Example 2

[0132] The anti-inflammatory effects of several ivy saponin derivatives prepared in Examples 1-7 were tested, and the specific testing methods are as follows:

[0133] 1.2 Griess method for detecting NO levels

[0134] In vivo, nitrogen (NO) is produced from L-arginine (L-Arg) catalyzed by nitric oxide synthases (NOS). After NOS production, it can act on its own cells or diffuse to neighboring cells, binding to target receptors such as transcription factors and protein kinases, thereby exerting a series of regulatory effects. At sites of inflammation, NO acts on vascular smooth muscle cells, increasing their cGMP levels, causing vasodilation and increased permeability, facilitating the delivery of inflammatory mediators and pain-inducing substances to the site of action. It also increases the infiltration of monocytes into the inflammatory site during the inflammatory response. In the pathogenesis of immune-mediated inflammation, elevated NO levels can affect the pathological process through multiple pathways. For example, at high concentrations, NO activates NF-κB, inducing the production of pro-inflammatory cytokines such as TNF-α and IL-1. In various inflammatory or immune-mediated animal models, significantly increased NO concentrations and inhibition of NO synthesis can improve typical inflammatory symptoms.

[0135] RAW264.7 cells in the logarithmic growth phase were harvested, and the cell density was adjusted to 5 × 10⁶ cells per 100 μL. 4A suspension of cells was seeded into 96-well plates and incubated in a cell culture incubator (37°C, 5% CO2) for 24 h. Various compounds with cell viability close to 90% (hederone derivatives prepared in Examples 1-5 and Example 7) were selected. The hederone derivatives were diluted to the required concentration using culture medium and added to the cell culture medium in the corresponding wells, resulting in a concentration of 20 μM / mL. The same volume of blank culture medium was added to the wells of the Control and Model groups. The cells were incubated at CO2 for 2 h. Except for the Control group, all other groups were stimulated with 20 μL of LPS (1 μg / mL) and incubated at CO2 for 24 h. NO levels were detected using the Griess method. After 24 hours of cell culture, the supernatant was transferred to a new 96-well plate, 100 μL per well. An equal volume of Griess A and Griess B (Griess A solution: 0.5 g p-aminobenzenesulfonic acid dissolved in 150 mL of 10 wt% dilute acetic acid; Griess B solution: 0.1 g α-naphthylamine, 20 mL distilled water, and 150 mL of 10 wt% dilute acetic acid) was mixed, and 100 μL of the mixture was added to each well. The plate was then shaken for 10 min. OD values ​​were measured at 540 nm. Results are shown below. Figure 2 As shown. (Compared to the Control group:) ### P<0.001, compared with the Model group: *** P<0.001).

[0136] Experimental Example 3

[0137] The effects of Example 3 and the STING-specific inhibitor C-176 on NO production in RAW264.7 cells were investigated using the following methods:

[0138] RAW264.7 cells in the logarithmic growth phase were harvested, and the cell density was adjusted to 5 × 10⁶ cells per 100 μL. 4A suspension of cells was seeded into 96-well plates and incubated in a cell culture incubator (37°C, 5% CO2) for 24 h. Example 3 was diluted to the required concentration with culture medium, and C-176 was diluted to 1 μM with culture medium and added to the cell culture medium in the corresponding wells of the plates. The same volume of blank culture medium was added to the wells of the Control and Model groups. The cells were incubated at CO2 temperature for 2 h. Except for the Control group, all other groups were stimulated with 20 μL of LPS (1 μg / mL) and incubated at CO2 temperature for 24 h. NO levels were detected using the Griess method. After 24 hours of cell culture, the supernatant was transferred to a new 96-well plate, 100 μL per well. An equal volume of Griess A and Griess B (Griess A solution: 0.5 g p-aminobenzenesulfonic acid dissolved in 150 mL of 10 wt% dilute acetic acid; Griess B solution: 0.1 g α-naphthylamine, 20 mL distilled water, and 150 mL of 10 wt% dilute acetic acid) was mixed, and 100 μL of the mixture was added to each well. The plate was then shaken for 10 min. OD values ​​were measured at 540 nm. Results are shown below. Figure 2 As shown. (Compared to the Control group:) ### P<0.001, compared with the Model group: *** P<0.001).

[0139] Test results as follows Figure 3 As shown, the test results indicate that its anti-inflammatory activity at 20 μM / mL is similar to that of the C-176STING inhibitor. However, the preparation method of this invention is simpler, the raw materials are readily available, and the drug production cost is reduced.

[0140] As demonstrated in Examples 1-3, this invention modifies the hederoneinogen by linking (R)-(+)-1,2-dithiopentane-3-pentanoic acid at the C-23 position using different linking chains, thus obtaining a new chemical entity. Hederoneinogen itself is cytotoxic, with a cell survival rate of 60% at a concentration of 40 μM / mL. The structural modification of this invention allows the hederoneinogen to exhibit both high anti-inflammatory activity and lower cytotoxicity. Considering the drug's inhibitory activity against NO at different concentration gradients, Example 3 exhibits the best anti-inflammatory potential, achieving an inhibition rate of 81.70% against NO at a concentration of 20 μM. The preparation method of the hederoneinogen derivative disclosed in this invention is relatively simple, using readily available raw materials, and can be used for large-scale production of hederoneinogen derivatives. The prepared hederoneinogen derivative has broad application prospects in the treatment of anti-inflammatory drugs and as a pharmaceutical carrier.

[0141] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. An ivy saponin derivative, characterized in that, The general formula of the ivy saponin derivatives is shown in Formula I: R1 includes , , , , and One of them; R2 is ; In R1, -NH- is connected to R2.

2. The use of the ivy saponin derivative of claim 1 in the preparation of anti-inflammatory drugs.

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