Cordycepin-modified glycyrrhetinic acid derivative as well as preparation method and application thereof

Abstract

The invention relates to the technical field of organic synthesis, and particularly discloses a cordycepin-modified glycyrrhetinic acid derivative as well as a preparation method and application thereof. The invention provides the synthetic method of the cordycepin-modified glycyrrhetinic acid derivative, so that the yield of the target compound glycyrrhetinic acid derivative is good, meanwhile, the novel structure has multiple potential biological activities, and the chemical space of the glycyrrhetinic acid derivative is widened. Meanwhile, the cordycepin-modified glycyrrhetinic acid derivative disclosed by the invention can improve ethanol-induced alcoholic liver injury of mice in a synergistic interaction manner, so that the cordycepin-modified glycyrrhetinic acid derivative can be effectively applied to treatment of clinical alcoholic liver injury.

Description

Technical Field

[0001] This invention relates to the field of organic synthesis technology, specifically to a cordycepin-modified glycyrrhetinic acid derivative, its preparation method, and its application in the preparation of products for preventing and treating alcoholic liver injury. Background Technology

[0002] Alcoholic liver disease (ALD) is a liver disease caused by long-term heavy alcohol consumption. It typically begins as alcoholic fatty liver, but can gradually progress to alcoholic hepatitis, liver fibrosis, and cirrhosis. The pathogenesis of ALD mainly includes the toxic effects of ethanol and its metabolites, disruption of the intestinal barrier function, oxidative stress, lipid peroxidation, inflammatory responses, and immune damage. According to the World Health Organization's global status report on alcohol and health, approximately 3 million people died from alcohol abuse in 2016, accounting for 5.3% of all deaths worldwide. The incidence of ALD in my country is also rising annually, with 700,000 deaths annually due to alcohol consumption, making it a significant global public health issue. Current treatments mainly include abstinence from alcohol, nutritional supplementation, and drug therapy; however, drug therapy suffers from low safety and significant side effects. Therefore, finding safe and effective natural remedies has become a focus of research for many scholars.

[0003] Glycyrrhetinic acid (GA) is a hydrolysis product of glycyrrhizic acid, the main active ingredient in licorice, and possesses various pharmacological activities, including anti-inflammatory, antiviral, antioxidant, hepatoprotective, and immunomodulatory effects. By modifying the structure of glycyrrhetinic acid (such as altering the functional groups like hydroxyl, carboxyl, and double bonds), a series of derivatives can be synthesized to enhance its biological activity, reduce toxic side effects, or improve pharmacokinetic properties. The core structure of glycyrrhetinic acid is an oleanane-type pentacyclic triterpenoid. Common derivative types and activities include: C-3 esterification, such as glycyrrhetinic acid stearate and glycyrrhetinic acid methyl ester, which enhances lipid solubility and improves bioavailability; C-3 amidation, such as glycyrrhetinic acid-3-aminoglucosamide, which enhances anti-inflammatory activity; and 11-deoxyglycyrrhetinic acid, with the C-11 ketone group reduced to a hydroxyl group, which reduces mineralocorticoid-like side effects (such as hypertension). A glycosylation group linked at the C-3 position, such as glucuronic acid derivatives, improves water solubility and is suitable for oral administration.

[0004] Cordycepin, also known as 3'-deoxyadenosine, is a natural nucleoside active substance extracted from Cordyceps fungi such as Cordyceps militaris. It has a variety of biological activities, including anti-tumor, immunomodulatory, anti-inflammatory, and antioxidant effects. Summary of the Invention

[0005] Purpose of the invention: The technical problem to be solved by the present invention is to address the shortcomings of the prior art by providing a cordycepin-modified glycyrrhetinic acid derivative, its preparation method and application.

[0006] To solve the above-mentioned technical problems, the present invention discloses the following technical solution: In a first aspect, the present invention discloses a glycyrrhetinic acid derivative modified with cordycepin as shown in Formula I, or a pharmaceutically acceptable salt thereof.

[0007] I R1 is selected from hydroxyl or acetoxy.

[0008] In a second aspect, the present invention discloses a method for preparing the cordycepin-modified glycyrrhetinic acid derivative described in the first aspect above.

[0009] The method includes: (1) Glycyrrhetinic acid 1 was acylated to prepare carboxylic acid compound 2; (2) The chlorination reaction of carboxylic acid compound 2 is used to prepare acyl chloride compound 3; (3) Acyl chloride compound 3 was condensed with cordycepin 4 to obtain cordycepin-modified glycyrrhetinic acid derivative I-1; (4) The hydroxyl groups of the cordycepin-modified glycyrrhetinic acid derivative I-1 are acylated to obtain cordycepin-modified glycyrrhetinic acid derivative I-2. .

[0010] In step (1), a mixed solution of glycyrrhetinic acid, catalyst, acylation reagent, base and organic solvent is reacted; in some embodiments, glycyrrhetinic acid and catalyst are mixed and then replaced with an inert gas, followed by the addition of an organic solvent, acylation reagent and base to obtain the mixed solution.

[0011] In step (1), the catalyst for the reaction is 4-dimethylaminopyridine, the acylation reagent is acetic anhydride, and the reaction further includes a base and an organic solvent; in some embodiments, the base is triethylamine; in some embodiments, the organic solvent includes dichloromethane; in some embodiments, the organic solvent is a combination of dichloromethane and N,N dimethylformamide, and in some embodiments, the volume ratio of dichloromethane to N,N dimethylformamide is 13-19:1, such as 16:1.

[0012] In step (1), the molar ratio of glycyrrhetinic acid, catalyst, acylation reagent and base is 1.0:0.05-0.15:3.5-4.5:2.5-3.5, and in some embodiments it is 1.0:0.1:4.0:3.0.

[0013] In step (1), the molar volume ratio of glycyrrhetinic acid to organic solvent is 0.08-0.16 mmol / mL, such as 0.12 mmol / mL.

[0014] In step (1), the reaction temperature is 20-26°C, and in some embodiments it is room temperature.

[0015] In step (1), the reaction is carried out until the raw materials are completely consumed, and in some embodiments, the reaction takes 3-5 hours.

[0016] In step (1), after the reaction is completed, the product is purified and separated in a silica gel column, concentrated, and carboxylic acid compound 2 is obtained; in some embodiments, the solvent for the silica gel column purification is dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is 13-17:1, such as 15:1.

[0017] In step (1), in some embodiments, glycyrrhetinic acid and 4-dimethylaminopyridine are added to the reactor, and dichloromethane and N,N-dimethylformamide are added under a nitrogen atmosphere and magnetic stirring to clarify the reaction solution. Then, acetic anhydride and triethylamine are added, and the reaction is stirred at room temperature. After the reaction is completed, the resulting product is purified and separated in a silica gel column.

[0018] In step (2), the reaction is carried out under anhydrous and oxygen-free conditions.

[0019] In step (2), the organic solvent for the reaction is a mixed solvent of ultra-dry toluene and thionyl chloride in a volume ratio of 1:0.5-1.5. In some embodiments, the volume ratio of the two is 1:1.

[0020] In step (2), the molar volume ratio of the carboxylic acid compound 2 to the organic solvent is 1 mmol: 24-32 mL, such as 1 mmol: 28 mL.

[0021] In step (2), the reaction temperature is 20-26°C, and in some embodiments it is room temperature.

[0022] In step (2), the reaction is carried out until the reaction is complete, and in some embodiments, the reaction is carried out for 10-14 hours.

[0023] In step (2), after the reaction is completed, excess solvent is removed by rotary evaporation, and toluene is added to concentrate under reduced pressure to wash away the residue.

[0024] In step (2), in some embodiments, carboxylic acid compound 2 is placed in a reactor and nitrogen is introduced. Ultra-dry toluene and thionyl chloride are added under an anhydrous and oxygen-free nitrogen atmosphere. The mixture is stirred overnight at room temperature. After the reaction is complete, excess thionyl chloride is removed by rotary evaporation. Finally, toluene is added and the residue is removed by multiple vacuum concentrations.

[0025] In step (3), the molar ratio of the acyl chloride compound 3 to cordycepin 4 is 1:0.5-1.5, and in some embodiments it is 1:1.

[0026] In step (3), the solvent for the reaction is pyridine.

[0027] In step (3), the molar volume ratio of cordycepin to solvent is 1 mmol: 95-105 mL, such as 1 mmol: 100 mL.

[0028] In step (3), the reaction temperature is 20-26°C, and in some embodiments it is room temperature.

[0029] In step (3), the reaction is carried out until the reaction is complete, and in some embodiments the reaction is carried out for 22-26 hours.

[0030] In step (3), under the protection of an inert gas, pyridine is added to the mixture of acyl chloride compound 3 and cordycepin 4 in an ice-water bath, and the temperature is raised to 20-26℃ for reaction.

[0031] In step (3), after the reaction is completed, the product is concentrated by column chromatography to obtain cordycepin-modified glycyrrhetinic acid derivative I-1; in some embodiments, the solvent for column chromatography is dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is 8-12:1, such as 10:1.

[0032] In step (3), in some embodiments, cordycepin and acyl chloride compound 3 are placed in a reactor and nitrogen is replaced. Pyridine is added under anhydrous and oxygen-free conditions and nitrogen protection in an ice-water bath. The temperature is gradually increased to room temperature for reaction. After the reaction is completed, the concentrated residue is purified by column chromatography.

[0033] In step (4), an acylation reagent is added to a mixed solution of cordycepin-modified glycyrrhetinic acid derivative I-1, a base, a catalyst, and an organic solvent to carry out the reaction.

[0034] In step (4), the catalyst for the reaction is 4-dimethylaminopyridine (DMAP), the acylation reagent is acetic anhydride, and the reaction also includes a base and an organic solvent; in some embodiments, the base is triethylamine; in some embodiments, the organic solvent is acetonitrile.

[0035] In step (4), the molar ratio of the cordycepin-modified glycyrrhetinic acid derivative I-1, the catalyst, the acylation reagent and the base is 1.0:0.10-0.18:3.2-4.2:1.5-2.5, and in some embodiments it is 1.0:0.14:3.7:2.0.

[0036] In step (4), the molar volume ratio of the cordycepin-modified glycyrrhetinic acid derivative I-1 to the organic solvent is 0.3 mmol: 1.5-2.5 mL, such as 0.3 mmol: 2 mL.

[0037] In step (4), the reaction temperature is 55-65°C, and in some embodiments it is 60°C.

[0038] In step (4), the reaction is carried out until the raw materials are completely consumed, and in some embodiments, the reaction is carried out for 6-10 hours.

[0039] In step (4), after the reaction is completed, the volatiles are removed by vacuum concentration, and the concentrated residue is purified by column chromatography to obtain the cordycepin-modified glycyrrhetinic acid derivative I-2; in some embodiments, the solvents for the silica gel column purification are dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is 8-12:1, such as 10:1.

[0040] In step (4), in some embodiments, the cordycepin-modified glycyrrhetinic acid derivative I-1, triethylamine and DMAP are added to acetonitrile and stirred. Acetic anhydride is slowly added dropwise, the reaction is heated, and the mixture is concentrated under reduced pressure after the reaction is completed. The concentrated residue is then purified by column chromatography.

[0041] Thirdly, the present invention discloses a pharmaceutical composition.

[0042] The pharmaceutical composition comprises (i) the cordycepin-modified glycyrrhetinic acid derivative described in the first aspect above, or a pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier.

[0043] The content of component (i) accounts for 1%-99% of the total mass of the pharmaceutical composition.

[0044] Fourthly, the present invention discloses the use of the cordycepin-modified glycyrrhetinic acid derivative described in the first aspect above, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the third aspect, in the preparation of products for treating alcoholic liver injury.

[0045] Compared with the prior art, the present invention has the following beneficial effects:

[0046] This invention provides a method for synthesizing cordycepin-modified glycyrrhetinic acid derivatives, achieving good yields of the target compound, glycyrrhetinic acid derivatives. For example, the total yield of cordycepin-modified glycyrrhetinic acid derivative I-1 is as high as 10.72%, far superior to the 5.4% of the prior art. This invention further modifies compound I-1 by acetylation of the hydroxyl group, ultimately obtaining cordycepin-modified glycyrrhetinic acid derivative I-2 in a high yield, with a total yield of up to 9.1%.

[0047] This invention provides glycyrrhetinic acid derivatives modified with cordycepin that possess a variety of potential biological activities, thus broadening the chemical space of glycyrrhetinic acid derivatives.

[0048] The cordycepin-modified glycyrrhetinic acid derivative of this invention can improve ethanol-induced alcoholic liver injury in mice through a synergistic effect, and thus can be effectively applied to the clinical treatment of alcoholic liver injury. Attached Figure Description

[0049] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments, and the advantages of the present invention in the above and / or other aspects will become clearer.

[0050] Figure 1 This is one of the reaction pathways of the present invention.

[0051] Figure 2 This is one of the reaction pathways of the present invention.

[0052] Figure 3 The image shows the 1H NMR spectrum of glycyrrhetinic acid derivative I-1 modified with cordycepin.

[0053] Figure 4 This is the carbon NMR spectrum of glycyrrhetinic acid derivative I-1 modified with cordycepin.

[0054] Figure 5 The image shows the 1H NMR spectrum of glycyrrhetinic acid derivative I-2 modified with cordycepin.

[0055] Figure 6 The image shows the carbon NMR spectrum of glycyrrhetinic acid derivative I-2 modified with cordycepin. Detailed Implementation

[0056] The present invention can be better understood from the following embodiments. However, those skilled in the art will readily understand that the descriptions in the embodiments are for illustrative purposes only and should not, and will not, limit the invention as detailed in the claims.

[0057] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; unless otherwise specified, the reagents and materials are commercially available.

[0058] The reagents used in this invention were prepared with reference to relevant literature, and the solvents were purified and refined.

[0059] Table 1. Experimental reagents required for the experiment

[0060] Comparative Example 1

[0061] In the early stages of the experiment, the plan was to use a condensation reagent to promote the reaction between the carboxyl group of glycyrrhetinic acid and the amino group of cordycepin, aiming to form an amide bond by removing a water molecule, thereby synthesizing the target compound. However, the experimental process revealed that the results were not ideal, so the reaction conditions were simply screened.

[0062] Table 2 Effects of different solvents on the experiment

[0063] Cordycepin exhibits poor solubility (Table 2). Items 1 and 4 show that cordycepin has poor solubility in DCM and CH3CN, while items 2, 3, and 5 indicate better solubility in DMF, DCM+DMF, and Py. Compared to cordycepin, glycyrrhetinic acid has better solubility and can dissolve in most solvents. However, literature review indicates that the activated ester formed by the reaction of glycyrrhetinic acid with HOBt reacts more readily in high-boiling-point solvents (except DMF), and heating also increases the ammonolysis rate. Therefore, pyridine was ultimately chosen as the solvent.

[0064] Table 3. Effects of different catalysts and bases on the experiment

[0065] In the condensation experiment, the addition of a base and a catalyst effectively prevented racemization of the chiral center and increased the condensation reaction rate. As can be seen from items 1 to 4 (Table 3), changing the combination of base and catalyst had no significant effect on the reaction results; therefore, the combination of Et3N and DMAP was subsequently used in the experiment.

[0066] Table 4. Effects of different condensing agent combinations on the experiment.

[0067] As can be seen from items 1 to 5, different combinations of condensing agents had no significant impact on the experimental results (Table 4), but all showed instances where the raw materials were not consumed. LCMS results indicated a large amount of cordycepin remaining; at RT=9.37, MW=704.36, suggesting the formation of the target product, but in very small quantities. This was likely due to insufficient reaction time and temperature, so the reaction time was extended from 24 h to 48 h. Reflux heating at 110℃ was also attempted, but the results showed almost no significant change. Considering the potential influence of the glycyrrhetinic acid and cordycepin hydroxyl groups on this experiment, the reactants were then screened.

[0068] Table 5. Effects of different reactants on the experiment

[0069] According to items 1 to 5, it can be seen that changing the reactants in the experiment (Table 5) did not result in any significant difference. Among them, glycyrrhizic acid directly condensed with cordycepin, and the raw materials were basically not consumed. In the remaining combinations of raw materials, cordycepin was largely remaining, and glycyrrhetinic acid remained at the stage of activation to ester. No effective condensation reaction occurred in this experiment.

[0070] Despite numerous attempts, including changing the solvent, condensing agent, and catalyst, and adjusting the reaction temperature and time, the results were unsatisfactory. This prompted further optimization of experimental conditions and exploration of alternative strategies to achieve the desired ligation product.

[0071] Example 1 Glycyrrhetinic acid (5 g, 10.62 mmol) and 4-dimethylaminopyridine (129.5 mg, 1.06 mmol) were placed in a 250 mL round-bottom flask and purged with nitrogen. Under nitrogen atmosphere and magnetic stirring, 80 mL of dichloromethane and 5 mL of N,N dimethylformamide were added to clarify the solution. Then, acetic anhydride (4.1 mL, 43.7 mmol) and triethylamine (4.4 mL, 31.6 mmol) were added, and the mixture was stirred at room temperature for 4 h. TLC monitoring showed that the starting material had been completely consumed. The target product was purified and separated by silica gel column chromatography (dichloromethane:methanol = 15:1), concentrated, and evaporated to dryness to obtain carboxylic acid 2 (1.1 g, yield 20%). Subsequent products could be used directly without further purification.

[0072] Carboxylic acid 2 (0.5 g, 1.0 mmol) was placed in a 250 mL round-bottom flask and purged with nitrogen. Under an anhydrous and oxygen-free nitrogen atmosphere, 14 mL of ultra-dry toluene and 14 mL of thionyl chloride (toluene: thionyl chloride = 1:1) were added, and the mixture was stirred overnight at room temperature. LC-MS results showed RT = 13.89 and MW = 531.17, confirming the formation of intermediate product 3. After the reaction was complete, excess thionyl chloride was removed by rotary evaporation. Finally, 30 mL of toluene was added each time, and the mixture was concentrated under reduced pressure multiple times to remove any residue. The resulting product was used directly. Cordycepin (0.25 g, 1.0 mmol) and product 3 were placed in a 250 mL round-bottom flask and purged with nitrogen. Under an anhydrous and oxygen-free nitrogen atmosphere in an ice-water bath, 100 mL of pyridine was added, and the mixture was gradually heated to room temperature for 24 h. TLC (DCM: MeOH = 15:1) showed that cordycepin was completely consumed. The concentrated residue was purified by column chromatography (DCM: MeOH = 10:1), concentrated and evaporated to dryness to give a white solid compound I-1 (0.4 g, yield 53.6%). NMR was performed as follows: Figure 3 and Figure 4 As shown.

[0073] 1 H NMR (400 MHz, CDCl3) δ 8.30 (s, 1H), 8.05 (s, 1H), 6.03 (s, 2H), 5.94 (d, J = 3.2 Hz, 1H), 5.64 (s, 1H), 4.90 (m, 1H), 4.81 (m, 1H), 4.53 (dd, J =11.6, 4.9 Hz, 1H), 4.41 (dd, J = 12.1, 5.9 Hz, 1H), 4.27 (dd, J = 12.1, 3.3 Hz,1H), 2.83–2.75 (m, 1H), 2.48 (s, 1H), 2.36–2.22 (m, 3H), 2.07 (s, 3H), 2.03–1.53 (m, 10H), 1.47–1.27 (m, 9H), 1.17–0.98 (m, 12H), 0.89 (s, 6H), 0.75 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ200.2, 176.2, 171.1, 169.4, 155.6, 152.6, 148.7, 138.6, 128.4, 120.1, 93.0, 80.6, 78.4, 75.5, 65.5, 61.9, 55.0, 48.2, 46.0, 44.2, 43.2, 41.0, 38.8, 38.0, 37.6, 37.0, 34.0, 32.7, 31.8, 31.1, 28.5, 28.3, 28.0, 26.4, 26.3, 23.6, 23.3, 21.3, 18.7, 17.4, 16.7, 16.4. HRMSm / z (ESI)calcd for C 42 H 60 N5O7([M+H)) + ) 746.4493; found 746.4486. Example 2: Acylation

[0074] Cordycepin-modified glycyrrhetinic acid derivative I-1 (0.2 g, 0.3 mmol), triethylamine (0.1 mL, 0.7 mmol), and DMAP (5.0 mg, 0.041 mmol) were added to 2 mL of acetonitrile with stirring. Acetic anhydride (0.1 mL, 1.1 mmol) was slowly added dropwise, and the mixture was heated to 60 °C for 8 h. After the reaction was completed, the mixture was concentrated under reduced pressure to remove volatiles. The concentrated residue was purified by column chromatography (DCM: MeOH = 10:1) to give a white powdery solid compound I-2 (0.2 g, yield 84.6%). NMR was as follows. Figure 5 and Figure 6 As shown.

[0075] 1 H NMR (400 MHz, CDCl3) δ 8.31 (s, 1H), 7.99 (s, 1H), 6.07 (d, J = 1.7 Hz,1H), 5.99 (s, 2H), 5.89 (d, J = 6.2 Hz, 1H), 5.68 (s, 1H), 4.69 – 4.62 (m, 1H), 4.52 (dd, J = 11.6, 4.8 Hz, 1H), 4.39 (dd, J = 11.9, 7.2 Hz, 1H), 4.27 (dd, J=11.9, 3.1 Hz, 1H), 2.83 – 2.74 (m, 2H), 2.36 (s, 1H), 2.27 (d, J = 1.8 Hz, 1H), 2.15 (s, 3H), 2.06 (s, 3H), 1.95 – 1.52 (m, 10H), 1.43 – 1.24 (m, 9H), 1.18 –1.01 (m, 12H), 0.89 (d, J = 2.3 Hz, 6H), 0.65 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 200.3, 176.1, 171.1, 170.2, 169.5, 155.6, 153.0, 149.2, 139.9, 128.4, 120.2, 90.3, 80.6, 78.3, 65.5, 61.7, 55.0, 48.1, 45.4, 44.1, 43.1, 41.1, 38.8, 38.0, 37.5, 37.0, 33.4, 32.7, 31.7, 31.1, 28.3, 28.1, 28.0, 26.4, 23.6, 23.3, 21.3, 21.0, 18.6, 17.4, 16.7, 16.5. HRMSm / z (ESI) calcd for C 44 H 62 N5O8 ([M+H) + )788.4593; found 788.4598. Comparative Example 2 Glycyrrhetinic acid (5 g, 10.62 mmol) was placed in a reaction flask, and 200 mL of dichloromethane was added. Oxaloyl chloride (2.70 g, 21.24 mmol) was then added, followed by 53 μL of N,N-dimethylformamide. The reaction was carried out under reflux at 50 °C. After the reaction was completed, the reaction solution was distilled under reduced pressure, recrystallized from n-hexane, and filtered to obtain glycyrrhetinic chloride derivative (1.69 g, 3.19 mmol, yield 30.0%).

[0076] Under nitrogen protection, cordycepin (0.40 g, 1.60 mmol) and glycyrrhetinic acid (1.69 g, 3.19 mmol) were dissolved in 32 mL of anhydrous pyridine. Triethylamine (1.22 mL, 8.75 mmol) was added and reacted in one pot. The reaction was heated to 80 °C and monitored by high performance liquid chromatography until the reactant concentration stopped. The reaction solution was filtered, extracted with water and ethyl acetate, and the organic phase was collected and concentrated under reduced pressure to obtain a mixture. The concentrated mixture was purified by column chromatography (DCM: MeOH = 10:1) to give a white solid compound I-1 (0.21 g, yield 18%).

[0077] Application Example 1: Study on the effect of cordycepin-glycyrrhetinic acid derivative on alcoholic liver injury in mice 1. Experimental equipment and materials Table 6. Instruments and Equipment Required for the Experiment

[0078] Table 7 Experimental reagents required for the experiment

[0079] 2. Experimental Methods (1) Animal grouping and administration Male C57 mice, aged 6-8 weeks and weighing (18±2) g, were purchased from Nanjing Qinglongshan Animal Breeding Farm and housed at the Animal Center of Nanjing University of Technology. The mice were placed in a sterile environment with free access to food and water at 22-26 ℃ and 45-55% humidity, and underwent a 12 / 12-hour light / dark cycle. They were acclimatized for one week prior to the experiment and fed a Lieber-DeCarli diet. The mice were randomly divided into the following groups of 8 mice each: Control group (Contorl): From day 1 to day 10, participants had free access to Lieber-DeCarli feed without ethanol. On day 11, at 8:00 AM, participants were given 20 mL / kg of 45% maltodextrin solution, and samples were taken from the body at 5:00 PM on the same day.

[0080] Pathological model (Model) group: From day 1 to 10, the animals were given free access to Lieber-DeCarli feed containing 5% ethanol. On day 11, at 8:00 a.m., the animals were given 20 mL / kg of 31.5% ethanol solution, and the animals were dissected and smeared at 5:00 p.m. on the same day.

[0081] Cordycepin low (CL), medium (CM), and high (CH) dose groups: The rearing environment and modeling conditions for the three dose groups were the same as those for the model group. From day 1 to day 11 of the experiment, 2.5 mg / mL, 5.0 mg / mL, and 10.0 mg / mL of cordycepin physiological saline solutions were prepared daily. Cordycepin was administered by gavage daily at doses of 25 mg / kg, 50 mg / kg, and 100 mg / kg, respectively. Administration on day 11 was at 7:00 AM.

[0082] Cordycepin-glycyrrhetinic acid derivative low (CL), medium (CM), and high (CH) dose groups: The rearing environment and modeling conditions for the three dose groups were the same as those for the model group. From day 1 to day 11 of the experiment, 2.5 mg / mL, 5.0 mg / mL, and 10.0 mg / mL of cordycepin-glycyrrhetinic acid derivative I-1 physiological saline solutions were prepared daily. Cordycepin-glycyrrhetinic acid derivative I-1 was administered by gavage daily at doses of 25 mg / kg, 50 mg / kg, and 100 mg / kg, respectively. Administration on day 11 was at 7:00 AM.

[0083] Cordycepin-glycyrrhizic acid derivative group: The rearing environment and modeling conditions were the same as the model group. From day 1 to day 11 of the experiment, a 10.0 mg / mL solution of cordycepin-glycyrrhizic acid derivative was prepared daily, and the derivative was administered by gavage at a dose of 100 mg / kg daily. The administration time on the 11th day was 7:00 AM.

[0084] Silymarin (PC) group: The feeding environment and modeling method were the same as the model group. During the experiment from day 1 to day 11, 20.0 mg / mL silymarin saline solution was prepared daily, and silymarin was administered by gavage at a dose of 200 mg PC / kg daily. The administration time on day 11 was 7:00 AM.

[0085] (2) Liver injury assessment indicators Serum biochemical markers in mice: When the liver is damaged, it releases large amounts of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) into the serum. Furthermore, triglyceride (TG) levels in the blood also increase when the liver develops fatty liver disease. Therefore, a kit can be used to analyze ALT, AST, and TG in mouse serum to assess the degree of liver damage.

[0086] 3. Experimental Results As shown in Table 8, compared with the blank control group, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and triglycerides (TG) in the pathological model group mice were significantly increased. Compared with the pathological model group, all drug groups could significantly reduce the serum levels of ALT, AST, and TG in mice. Among them, cordycepin and cordycepin-glycyrrhetinic acid derivatives could alleviate the ethanol-induced elevation of ALT, AST, and TG in a dose-dependent manner; 100 mg / kg cordycepin had a more significant effect on reducing ALT and AST, while 100 mg / kg cordycepin-glycyrrhetinic acid derivatives had the most significant effect on reducing ALT, AST, and TG, especially the reduction effect of ALT and AST was much better than that of the silymarin group. The above results indicate that the cordycepin-glycyrrhetinic acid derivative of the present invention can significantly reduce the abnormal elevation of ALT, AST, and TG in the serum of mice with alcoholic liver injury, thereby improving ethanol-induced alcoholic liver injury in mice.

[0087] Table 8 Effects of cordycepin-glycyrrhetinic acid derivatives on serum markers in mice with alcoholic liver injury

[0088] In summary, the results of this invention show that cordycepin-glycyrrhetinic acid derivatives can reduce the abnormal elevation of ALT, AST and TG in the serum of mice with alcoholic liver injury, thereby improving ethanol-induced alcoholic liver injury in mice.

[0089] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. A glycyrrhetinic acid derivative modified with cordycepin as shown in Formula I, or a pharmaceutically acceptable salt thereof; I in, R1 is selected from hydroxyl or acetoxy.

2. A method for preparing the cordycepin-modified glycyrrhetinic acid derivative as described in claim 1, characterized in that, include: (1) Glycyrrhetinic acid 1 was acylated to prepare carboxylic acid compound 2; (2) The chlorination reaction of carboxylic acid compound 2 is used to prepare acyl chloride compound 3; (3) Acyl chloride compound 3 was condensed with cordycepin 4 to obtain cordycepin-modified glycyrrhetinic acid derivative I-1; 。 3. The preparation method according to claim 2, characterized in that, In step (1), the catalyst for the reaction is 4-dimethylaminopyridine, the acylation reagent is acetic anhydride, and the reaction further includes a base and an organic solvent; optionally, the base is triethylamine, and the organic solvent includes dichloromethane; optionally, the molar ratio of glycyrrhetinic acid, catalyst, acylation reagent and base is 1.0:0.05-0.15:3.5-4.5:2.5-3.5; optionally, the molar volume ratio of glycyrrhetinic acid and organic solvent is 0.08-0.16 mmol / mL; optionally, the reaction temperature is 20-26℃.

4. The preparation method according to claim 2, characterized in that, In step (2), the reaction is carried out under anhydrous and oxygen-free conditions; optionally, the organic solvent for the reaction is a mixed solvent of ultra-dry toluene and thionyl chloride in a volume ratio of 1:0.5-1.5; optionally, the molar volume ratio of the carboxylic acid compound 2 to the organic solvent is 1 mmol:24-32 mL; optionally, the temperature of the reaction is 20-26℃.

5. The preparation method according to claim 2, characterized in that, In step (3), the molar ratio of the acyl chloride compound 3 to cordycepin 4 is 1:0.5-1.5; optionally, the solvent for the reaction is pyridine; optionally, the molar volume ratio of cordycepin to solvent is 1 mmol:95-105 mL; optionally, the reaction temperature is 20-26℃.

6. The preparation method according to claim 2, characterized in that, include: The cordycepin-modified glycyrrhetinic acid derivative I-1 was subjected to an acylation reaction to obtain cordycepin-modified glycyrrhetinic acid derivative I-2. 。 7. The preparation method according to claim 6, characterized in that, The catalyst for the reaction is 4-dimethylaminopyridine, the acylation reagent is acetic anhydride, and the reaction further includes a base and an organic solvent; optionally, the base is triethylamine, and the organic solvent is acetonitrile; optionally, the molar ratio of the cordycepin-modified glycyrrhetinic acid derivative I-1, the catalyst, the acylation reagent, and the base is 1.0:0.10-0.18:3.2-4.2:1.5-2.5; optionally, the molar volume ratio of the cordycepin-modified glycyrrhetinic acid derivative I-1 to the organic solvent is 0.3 mmol:1.5-2.5 mL; optionally, the reaction temperature is 55-65℃.

8. A pharmaceutical composition, characterized in that, Includes (i) the cordycepin-modified glycyrrhetinic acid derivative of claim 1, or a pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier.

9. The pharmaceutical composition according to claim 8, characterized in that, The content of component (i) is 1%-99% of the total mass of the pharmaceutical composition.

10. The use of the cordycepin-modified glycyrrhetinic acid derivative of claim 1, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 8 or 9, in the preparation of an anti-alcoholic liver injury product.

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