Preparation method of 20 (Z)-ginsenoside F4 and application of 20 (Z)-ginsenoside F4 in preparation of anti-liver injury medicine
The preparation of 20(Z)-ginsenoside F4 from ginsenoside Re via acid-catalyzed conversion solves the problem of the difficulty in preparing rare ginsenosides, providing a highly effective hepatoprotective drug component suitable for the treatment of liver damage caused by chemotherapy drugs.
Patent Information
- Application Number
- CN202511123561.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-12
- Publication Date
- 2025-11-14
AI Technical Summary
Existing technologies make it difficult to efficiently prepare rare ginsenoside 20(Z)-ginsenoside F4, and its application in the treatment of liver damage caused by chemotherapy drugs is limited.
20(Z)-ginsenoside F4 was prepared from ginsenoside Re by acid-catalyzed conversion. High-purity 20(Z)-ginsenoside F4 was obtained by heteropolyacid catalysis, ultrasonic treatment, extraction and high-performance liquid chromatography purification.
We have achieved efficient, safe and environmentally friendly preparation of 20(Z)-ginsenoside F4, which has significant hepatoprotective activity and is suitable for preparing drugs to treat liver damage caused by chemotherapy.
Smart Images
Figure CN120939031A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of biomedicine, and in particular to a method for preparing 20(Z)-ginsenoside F4 and its application in the preparation of drugs for treating liver damage. Background Technology
[0002] In the clinical treatment of malignant tumors, the incidence and mortality rates of common cancers such as lung cancer, breast cancer, colorectal cancer, and prostate cancer are rapidly increasing, posing a serious threat to human health. Chemotherapy, as one of the mainstream and effective treatments for malignant tumors, plays an irreplaceable role in slowing tumor progression and prolonging the survival of patients with malignant tumors. However, the liver, as the core organ for drug metabolism in the human body, is continuously exposed to chemotherapeutic drugs and their active / toxic metabolites during chemotherapy, making it one of the most vulnerable target organs.
[0003] Chemotherapy-induced liver injury (CILI) has become a high-incidence and serious complication in cancer treatment. It directly hinders the successful implementation of chemotherapy regimens, reduces treatment efficacy, and significantly worsens patients' quality of life. The pathogenesis of CILI is complex and diverse, mainly involving: direct cytotoxic effects (such as the damage of hepatocyte membranes to alkylating agents), metabolic activation pathways (such as the conversion of prodrugs into hepatotoxic substances by CYP450 enzyme systems), mitochondrial dysfunction (such as the interference of the respiratory chain by fluorouracil), inhibition of bile acid transporters (such as cholestasis caused by irinotecan), and immune-mediated inflammatory responses (such as immune checkpoint inhibitor-associated hepatitis). Its clinical phenotypes can be divided into hepatocellular injury type (characterized by significantly elevated ALT), cholestatic type (marked by elevated ALP and GGT), and mixed type. Severe cases can progress to acute liver failure, endangering the patient's life. Epidemiological data clearly show that approximately 10%-20% of patients receiving chemotherapy experience varying degrees of liver damage, with certain chemotherapy drugs (such as methotrexate, oxaliplatin, and tyrosine kinase inhibitors) posing a significantly higher risk of liver injury. It is noteworthy that pre-existing liver disease (such as fatty liver, viral hepatitis, and liver metastases), genetic susceptibility, and drug interactions all significantly increase the risk of clinically induced liver injury (CILI). Because CILI is often unpredictable and difficult to diagnose clinically, coupled with insufficient attention paid to it in clinical practice, its incidence is often underestimated or ignored. Despite its prevalence in clinical practice, there are currently no specific drugs available for its treatment, and this technological gap represents a key bottleneck restricting the improvement of chemotherapy efficacy in cancer treatment.
[0004] Ginsenosides, as the main active substances in ginseng, can be classified into protopanaxadiol type (such as Rb1, Rb2, Rc), protopanatriol type (such as Re, Rf, Rg1), octitilon type (such as ginsenoside F11), and other rare types (such as Rg5, Rh4, Rk3) based on differences in the aglycone skeleton and the position and number of glycosyl substitutions. Ginsenosides have a wide range of pharmacological activities, and in recent years, the organ-protective effects of ginseng and its saponin components have become a hot topic in the field of pharmaceutical research. Among them, 20(Z)-ginsenoside F4 is a rare ginsenoside component with unique activity, but it can only be isolated from ginseng leaves to date, and the content is extremely low (only about 0.05%), resulting in high acquisition costs and severely limiting its in-depth research and practical application in the field of medicine and health.
[0005] In existing technologies, the conversion and preparation methods for rare ginsenosides mainly include physical methods (such as high-pressure cooking and microwave conversion), biological methods (microbial conversion and enzymatic conversion), and chemical methods (acid degradation and alkali degradation). Although these methods each have their applicable scenarios, they also have obvious drawbacks and can only be applied to the preparation of some rare ginsenoside monomers. 20(Z)-ginsenoside F4, as an isomer of 20(E)-ginsenoside F4, is extremely difficult to obtain due to the significant difference in thermodynamic stability between the two during their formation. To date, there are no reports on its effective conversion and preparation in existing technologies, and this technical difficulty further exacerbates the current limited application status. Summary of the Invention
[0006] The purpose of this invention is to provide a method for preparing 20(Z)-ginsenoside F4 and its application in the preparation of drugs for treating liver damage, thereby addressing the problems existing in the prior art. This invention purifies and prepares the rare ginsenoside 20(Z)-ginsenoside F4 monomer from ginsenoside Re using an acid-catalyzed conversion method. This monomer exhibits significant advantages and broad prospects in the development of hepatoprotective drugs. It possesses strong hepatoprotective activity and aligns with the development trend of monomeric compound formulations, providing a highly efficient active ingredient for hepatoprotective drugs.
[0007] To achieve the above objectives, the present invention provides the following solution:
[0008] This invention provides the application of 20(Z)-ginsenoside F4 in the preparation of drugs for treating liver damage.
[0009] Furthermore, the liver injury mentioned is liver injury caused by chemotherapy drugs.
[0010] Furthermore, the chemotherapy drug is cisplatin.
[0011] This invention also provides a method for preparing 20(Z)-ginsenoside F4, comprising the following steps:
[0012] Ginsenoside Re was dissolved in a heteropoly acid aqueous solution and subjected to ultrasonic catalysis to obtain a conversion product solution.
[0013] The transformation product solution was extracted, and the resulting extract was concentrated under reduced pressure to obtain a degradation mixture of ginsenoside Re.
[0014] The degradation mixture of ginsenoside Re was purified by high performance liquid chromatography. The chromatographic peak eluent at 28.39 min was collected, concentrated and dried to obtain 20(Z)-ginsenoside F4.
[0015] Furthermore, the heteropolyacid is phosphotungstic acid, phosphotomolybdic acid, or silicotungstic acid.
[0016] Furthermore, the concentration of the heteropolyacid aqueous solution is 0.01-0.1 mol / L.
[0017] Furthermore, the ratio of the ginsenoside Re to the heteropolyacid aqueous solution is 1 mg:(3-5) mL.
[0018] Furthermore, the organic solvent used in the extraction process is dichloromethane.
[0019] Furthermore, the power of the ultrasonic catalytic treatment is 100-300W.
[0020] Furthermore, the chromatographic conditions for the purification process are as follows: mobile phase 70% methanol-0.1% formic acid water, isocratic elution, flow rate 3.5 mL / min, and detection wavelength 203 nm.
[0021] The present invention discloses the following technical effects:
[0022] This invention utilizes an acid-catalyzed conversion method to prepare and purify rare ginsenoside 20(Z)-ginsenoside F4 from ginsenoside Re, demonstrating significant advantages and broad prospects in the development of hepatoprotective drugs. The selected acidic catalyst is a solid heteropolyacid, which is recyclable after the catalytic reaction, achieving a yield of 16-21% for the conversion of ginsenoside Re to 20(Z)-ginsenoside F4. This method is green, efficient, and environmentally friendly. Furthermore, the preparation process is easy to operate, achieving high yields and high purity, and minimizing the introduction of toxic substances during production, ensuring product safety and avoiding toxic side effects and drug dependence. This monomer possesses strong hepatoprotective activity, aligning with the trend of monomeric compound formulation development and providing a highly effective active ingredient for hepatoprotective drugs.
[0023] This 20(Z)-ginsenoside F4, which combines high activity, excellent safety and environmentally friendly preparation characteristics, can be widely used in the preparation of hepatoprotective drugs or drug compositions, especially in the preparation of drugs or drug compositions for treating liver damage caused by chemotherapy drugs.
[0024] The preparation method provided by this invention ensures the feasibility of large-scale production of ginsenoside 20(Z)-ginsenoside F4, while its outstanding hepatoprotective efficacy meets the core clinical needs for hepatoprotective drugs. This comprehensively reflects the practical value of this invention in the field of drug development and provides a new and effective approach for the innovative development of hepatoprotective drugs. Attached Figure Description
[0025] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0026] Figure 1 The HPLC chromatogram of the degradation mixture of ginsenoside Re in Example 1 is shown below.
[0027] Figure 2 The HPLC chromatogram for the preparation of 20(Z)-ginsenoside F4 by separation and purification in Example 1 is shown below.
[0028] Figure 3 The HPLC chromatogram of 20(Z)-ginsenoside F4 in Example 1 is shown below.
[0029] Figure 4 The 1H NMR spectrum of 20(Z)-ginsenoside F4 in Example 1 ( 1 HNMR spectrum;
[0030] Figure 5 The carbon NMR spectrum of 20(Z)-ginsenoside F4 in Example 1 ( 13 C NMR spectrum;
[0031] Figure 6 The mass spectrum (MS) of 20(Z)-ginsenoside F4 in Example 1 is shown below.
[0032] Figure 7 A statistical graph showing the survival rate of human hepatocytes induced by different concentrations of cisplatin;
[0033] Figure 8 A statistical graph showing the survival rate of hepatocytes after intervention with different concentrations of 20(Z)-ginsenoside F4;
[0034] Figure 9The image shows the ALT level detection results of cisplatin-induced hepatocytes under the action of 20(Z)-ginsenoside F4.
[0035] Figure 10 The figure shows the results of AST level detection in cisplatin-induced hepatocytes under the action of 20(Z)-ginsenoside F4. Detailed Implementation
[0036] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.
[0037] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.
[0038] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.
[0039] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.
[0040] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.
[0041] This invention performs purity testing and structural analysis on 20(Z)-ginsenoside F4 samples under the following conditions:
[0042] Purity was determined by HPLC area normalization method. Chromatographic conditions: column: Zorbax Eclipse XDB C-18 (5μm; 4.6mm id×250mm); mobile phase: methanol-water (70:30 v / v); flow rate: 1.0mL / min; detection wavelength: 203nm; injection volume: 20μL.
[0043] Structural analysis was performed using an Orbitrap Elite mass spectrometer (Thermo Scientific, Bremen, Germany) and a Bruker DRX-500 nuclear magnetic resonance spectrometer (TMS as internal standard, C5D5N as solvent). The MS and NMR data of 20(Z)-ginsenoside F4 were obtained.
[0044] This invention utilizes acid conversion and high-performance liquid chromatography (HPLC) to obtain high-purity 20(Z)-ginsenoside F4 from the conversion product of ginsenoside Re. Pharmacological experiments, including an in vitro hepatoprotective assay against cisplatin-induced hepatocyte injury, demonstrated its strong hepatoprotective activity. The 20(Z)-ginsenoside F4 monomer obtained by this method exhibits high purity and stable quality, making it suitable for use in the development of hepatoprotective drugs or the preparation of pharmaceutical compositions.
[0045] Example 1
[0046] A method for preparing 20(Z)-ginsenoside F4 by catalytic conversion of ginsenoside Re, the specific steps of which are as follows:
[0047] (1) Catalytic transformation of ginsenoside Re
[0048] Weigh 50 mg of ginsenoside Re and dissolve it in 250 mL of 0.025 mol / L phosphotungstic acid aqueous solution. Place this solution in an ultrasonic cell extractor and react for 1 h at an ultrasonic power of 150 W and an ultrasonic temperature of 60 °C. After 1 h, cool in an ice bath to terminate the reaction and obtain a conversion product solution. Extract the conversion product solution twice with an equal volume of dichloromethane, combine the extracts, and concentrate under reduced pressure to obtain a degradation mixture of ginsenoside Re. The HPLC chromatogram of the degradation mixture of ginsenoside Re is shown below. Figure 1 .
[0049] (2) Purification and preparation of 20(Z)-ginsenoside F4
[0050] The degradation mixture of ginsenoside Re obtained in step (1) was freeze-dried to obtain 61.4 mg, dissolved in chromatographic methanol, filtered through a 0.22 μm filter membrane, and then purified by C18 high-performance liquid chromatography. The chromatographic conditions were: mobile phase 70% methanol-0.1% formic acid-water, isocratic elution, flow rate 3.5 mL / min, and detection wavelength 203 nm. The eluent of the chromatographic peak at 28.39 min was collected (see...). Figure 2 The solvent was recovered under reduced pressure, and the product was freeze-dried to obtain 20(Z)-ginsenoside F4 powder, weighing 6 mg. The conversion rate was calculated to be 12%.
[0051] Example 2
[0052] A method for preparing 20(Z)-ginsenoside F4 by catalytic conversion of ginsenoside Re, the specific steps of which are as follows:
[0053] (1) Catalytic transformation of ginsenoside Re
[0054] Weigh 50 mg of ginsenoside Re and dissolve it in 200 mL of 0.01 mol / L silicotungstic acid aqueous solution. Place this solution in an ultrasonic cell extractor and react for 0.5 h at an ultrasonic power of 100 W and an ultrasonic temperature of 80 °C. After 0.5 h, cool in an ice bath to terminate the reaction and obtain a conversion product solution. Extract the conversion product solution twice with an equal volume of dichloromethane, combine the extracts, and concentrate under reduced pressure to obtain a degradation mixture of ginsenoside Re.
[0055] (2) Purification and preparation of 20(Z)-ginsenoside F4
[0056] The degradation mixture of ginsenoside Re obtained in step (1) was freeze-dried to obtain 58.6 mg, dissolved in chromatographic methanol, filtered through a 0.22 μm filter membrane, and then purified by C18 high-performance liquid chromatography (HPLC). The chromatographic conditions were: mobile phase 70% methanol-0.1% formic acid-water, isocratic elution, flow rate 3.5 mL / min, and detection wavelength 203 nm. The eluent of the peak at 28.39 min was collected, the solvent was recovered under reduced pressure, and the eluent was freeze-dried to obtain 20(Z)-ginsenoside F4 powder, weighing 5 mg. The conversion rate was calculated to be 10%.
[0057] Example 3
[0058] A method for preparing 20(Z)-ginsenoside F4 by catalytic conversion of ginsenoside Re, the specific steps of which are as follows:
[0059] (1) Catalytic transformation of ginsenoside Re
[0060] Weigh 50 mg of ginsenoside Re and dissolve it in 150 mL of 0.1 mol / L phosphotungstic acid aqueous solution. Place this solution in an ultrasonic cell extractor and react for 1.5 h at an ultrasonic power of 300 W and an ultrasonic temperature of 40 °C. After 1.5 h, cool in an ice bath to terminate the reaction and obtain the conversion product solution. Extract the conversion product solution twice with an equal volume of dichloromethane, combine the extracts, and concentrate under reduced pressure to obtain the degradation mixture of ginsenoside Re.
[0061] (2) Purification and preparation of 20(Z)-ginsenoside F4
[0062] The degradation mixture of ginsenoside Re obtained in step (1) was freeze-dried to obtain 56.2 mg, dissolved in chromatographic methanol, filtered through a 0.22 μm filter membrane, and then purified by C18 high-performance liquid chromatography (HPLC). The chromatographic conditions were: mobile phase 70% methanol-0.1% formic acid-water, isocratic elution, flow rate 3.5 mL / min, and detection wavelength 203 nm. The eluent of the peak at 28.39 min was collected, the solvent was recovered under reduced pressure, and the eluent was freeze-dried to obtain 20(Z)-ginsenoside F4 powder, weighing 4 mg. The conversion rate was calculated to be 8%.
[0063] Example 4
[0064] (1) Purity testing
[0065] The purity of 20(Z)-ginsenoside F4 prepared in Example 1 was determined by high performance liquid chromatography (HPLC) to be 96.3%. Figure 3 Chromatographic conditions: Column: Zorbax Eclipse XDB C-18 (5 μm; 4.6 mm id × 250 mm); Mobile phase: Methanol:water (v / v) 70:30; Flow rate: 1.0 mL / min; Detection wavelength: 203 nm; Injection volume: 20 μL. Under these chromatographic conditions, the regression equation for the 20(Z)-ginsenoside F4 standard curve was y = 4.667x - 30.060, R0 2 =0.9993; good linearity was observed in the injection volume range of 0.2 μg to 30 μg.
[0066] (2) Structural Analysis
[0067] The purified product was structurally characterized using high-resolution mass spectrometry (HMS) and nuclear magnetic resonance (NMR) spectroscopy, and the MS and NMR spectral characteristics of 20(Z)-ginsenoside F4 were obtained. An Orbitrap Elite mass spectrometer (Thermo Scientific, Bremen, Germany) and a Bruker DRX-500 NMR spectrometer (TMS as internal standard, C5D5N as solvent) were used. The experimental results are analyzed below.
[0068] Analysis of high-resolution mass spectrometry results:
[0069] High-resolution mass spectrometry of 20(Z)-ginsenoside F4 as follows Figure 6 As shown in the figure. The m / z value in the spectrum is 765.4791 [MH]. - (Calculated value 765.4789; C) 42 H 69 O 12 This indicates that its molecular formula is C. 42 H 70 O 12 This is consistent with the results.
[0070] Analysis of results from nuclear magnetic resonance spectroscopy:
[0071] 20(Z)-Ginsenoside F4 is a white amorphous powder; the spots turn purple when developed with 5% sulfuric acid ethanol; the Liberman-Burchard reaction is positive.
[0072] Through the analysis of 20(Z)-ginsenoside F4 1 H NMR (C5D5N) ( Figure 4 )and 13 C NMR (C5D5N) Figure 5 The NMR data were analyzed and compared with the literature, and were assigned as shown in Table 1.
[0073] Table 120(Z)-Ginsenoside F4 1 HNMR and 13 CNMR data
[0074]
[0075]
[0076] The above test data confirms that the sample obtained in this invention is 20(Z)-ginsenoside F4, and its chemical structural formula is as follows:
[0077]
[0078] The present invention further demonstrates its hepatoprotective effect through the following pharmacodynamic experiments.
[0079] Example 5
[0080] The hepatoprotective effect of 20(Z)-ginsenoside F4 was determined using a cisplatin-induced in vitro hepatocyte injury model.
[0081] 1. Experimental Methods
[0082] (1) Model building
[0083] HL7702 cells in the logarithmic growth phase were seeded into 96-well plates and divided into control and experimental groups. The control group had a cisplatin concentration of 0 μM, while the experimental groups had cisplatin concentrations of 10, 15, 20, and 25 μM. Each group was initially cultured in 100 μL of RPMI-DMEM medium containing 10% fetal bovine serum for 24 h, then the medium was replaced with the corresponding concentration of cisplatin, and cultured for another 24 h. Cell culture medium containing 10% CCK-8 reagent was then added, and after 1 h, the absorbance was measured at 450 nm to calculate the survival rate of HL7702 cells stimulated by different concentrations of cisplatin.
[0084] (2) Detection of cell viability by fluorescence staining
[0085] Calcein AM (1000×), PI (1000×), and detection buffer were prepared into a Calcein AM / PI working solution at a ratio of 1:1:1000. Cisplatin-treated cells were treated with 0.625, 1.25, 2.5, and 5 μM 20(Z)-ginsenoside F4 for 24 h. The culture medium in the cell culture chamber was aspirated, and the cells were washed once with PBS. 50 μL of the Calcein AM / PI working solution was added to the lower culture chamber to act on the cells. The cells were incubated in the dark for 30 min and observed under an inverted fluorescence microscope.
[0086] (3) Detection of AST and ALT in hepatocyte supernatant
[0087] After HL7702 cells were treated with the appropriate drugs, cells from each group were collected. The collected cells were washed twice with phosphate buffer, centrifuged at 1000 r / min for 10 min, the supernatant was discarded, and the cell pellet was kept. Phosphate buffer was added, and the cells were sonicated under ice bath conditions. The protein concentration of the samples was determined according to the BCA kit, and then the AST and ALT kits were used.
[0088] 2. Experimental Results
[0089] The optimal concentration of cisplatin in the HL7702 liver injury model was determined to be 15 μM using the CCK8 assay. Figure 7 .
[0090] Quantitative results of cell viability analysis showed that, compared with the model group, the cell viability of 20(Z)-ginsenoside F4 at concentrations of 1.25 μM, 2.5 μM, and 5 μM was significantly higher than that of the model group. Furthermore, the cell viability gradually increased with increasing drug concentration, showing significant differences compared to the model group. This indicates that 20(Z)-ginsenoside F4 has a significant protective effect against cell damage in HL7702 cells. Figure 8 .
[0091] Cisplatin treatment significantly induced hepatocyte damage, leading to a significant increase in ALT and AST activities in the cell supernatant, with statistically significant differences. However, intervention with different doses of 20(Z)-ginsenoside F4 effectively inhibited the release of ALT and AST from cisplatin-treated cells and significantly reduced their activities in the supernatant, indicating that 20(Z)-ginsenoside F4 has a significant protective effect against cisplatin-induced hepatocyte damage. Figure 9 and Figure 10 .
[0092] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.
Claims
1. 20(Z)-Ginsenoside F4 in the preparation of drugs for treating liver damage.
2. The application according to claim 1, characterized in that, The liver injury mentioned is liver injury caused by chemotherapy drugs.
3. The application according to claim 2, characterized in that, The chemotherapy drug is cisplatin.
4. A method for preparing 20(Z)-ginsenoside F4, characterized in that, Includes the following steps: Ginsenoside Re was dissolved in a heteropoly acid aqueous solution and subjected to ultrasonic catalysis to obtain a conversion product solution. The transformation product solution was extracted, and the resulting extract was concentrated under reduced pressure to obtain a degradation mixture of ginsenoside Re. The degradation mixture of ginsenoside Re was purified by high performance liquid chromatography. The chromatographic peak eluent at 28.39 min was collected, concentrated and dried to obtain 20(Z)-ginsenoside F4.
5. The preparation method according to claim 4, characterized in that, The heteropolyacid is phosphotungstic acid, phosphomolybdic acid, or silicotungstic acid.
6. The preparation method according to claim 4, characterized in that, The concentration of the heteropolyacid aqueous solution is 0.01-0.1 mol / L.
7. The preparation method according to claim 4, characterized in that, The ratio of the ginsenoside Re to the heteropolyacid aqueous solution is 1 mg:(3-5) mL.
8. The preparation method according to claim 4, characterized in that, The organic solvent used in the extraction process is dichloromethane.
9. The preparation method according to claim 4, characterized in that, The power of the ultrasonic catalytic treatment is 100-300W.
10. The preparation method according to claim 4, characterized in that, The chromatographic conditions for the purification process were as follows: mobile phase 70% methanol-0.1% formic acid water, isocratic elution, flow rate 3.5 mL / min, and detection wavelength 203 nm.