CN103923158A - A-ring polyoxygenated glycyrrhetinic acid derivatives, preparation method and use thereof - Google Patents

Abstract

The invention discloses polyoxygenated substitution Enoxolone derivative of A ring and its preparation method and application,The general formula of the compound is as follows,Wherein R1 is selected from carbonyl or hydroxyl; R2,R3 be epoxy group between hydroxyl or R2 and R3; R4 be selected from-COOH,-COOR1,-CONH2,-CONHR1,-CONR1R2; R1,R2 be selected from the alkyl containing 1-15 carbon atom,Substituted or unsubstituted phenyl,Substituted or unsubstituted benzene alkyl,R1 may be the same or different with R2; Y is CH2 or C=O; 18 hydrogen is α or beta comfiguration; R1,R2,When R3 is hydroxyl,Its hydroxyl replaces configuration to be respectively α or beta comfiguration. The compound and its salt of structure novel provided by the present invention have stronger liver-protecting activity, can be used for the preparation of hepatic; And the purposes with antibacterial or resisting tobacco mosaic virus, good application prospect is all had in medicine and pesticide field.

Description

A-ring polyoxygenated glycyrrhetinic acid derivatives, preparation method and use thereof

technical field

The invention relates to A-ring polyoxidized substituted glycyrrhetinic acid derivatives, a preparation method and application thereof, and belongs to the technical field of medicinal chemistry.

Background technique

The liver is an important metabolic organ and barrier organ, and liver disease seriously threatens human health and life. Liver disease is a common and frequently-occurring disease. The pathogenic factors include viruses, bacteria, chemicals, alcohol, high-fat diet, etc., among which viral hepatitis is dominant. Chronic hepatitis B is the main cause of liver cirrhosis and liver cancer. According to statistics, there are about 130 million chronic hepatitis B virus carriers in my country, accounting for 10% of the country's total population; 25% to 40% of chronic hepatitis patients eventually die of liver cirrhosis or liver cancer; drug-induced hepatitis and alcoholic liver cirrhosis The incidence rate is also high. Therefore, the development of hepatoprotective drugs is of great significance.

Glycyrrhiza Root (Glycyrrhiza Root) was first recorded in "Shen Nong's Materia Medica", and 74 of the 110 prescriptions in "Treatise on Febrile Diseases" used licorice. It can be seen that licorice is an important traditional Chinese medicine. Glycyrrhiza licorice is flat and sweet in nature, and has the functions of neutralizing stress, nourishing the lungs, detoxifying, eliminating phlegm, relieving cough, dredging the meridian, benefiting qi and blood, and harmonizing various medicines (Kang Lei et al., 18β-glycyrrhetinic acid structure modification and research progress in biological activity , Chinese Herbal Medicine, 2012, 43(7): 1430-1442). The main active ingredients of licorice are glycyrrhizic acid and glycyrrhetinic acid. Glycyrrhizic acid is decomposed into glycyrrhetinic acid by gastric acid hydrolysis or liver β-glucuronidase in the human body, and then exerts its medicinal effect. Therefore, the medicinal effect of glycyrrhizic acid drugs is essentially determined by the medicinal effect of glycyrrhetinic acid (Gold Min et al. Research progress on the pharmacological effects of glycyrrhetinic acid, Medical Review, 2009, 15(11): 1712-1715).

Modern pharmacological studies have shown that glycyrrhetinic acid has various pharmacological activities such as anti-inflammatory, anti-ulcer, anti-virus (hepatitis virus, HIV, SARS virus, etc.), hypolipidemic, anti-tumor, liver protection, and antibacterial. Glycyrrhizin drugs are currently commonly used clinical liver protection drugs, and glycyrrhizic acid injection preparations are widely used in the treatment of hepatitis. Kimura Mitsutoshi et al. found that glycyrrhetinic acid is a hepatic cytokinin, which induces DNA synthesis in hepatocytes, promotes the growth of hepatocytes, and then exerts hepatoprotective effects (Kimura M, Inoue H, Kazuhiro Hirabayashi K. et al. Glycyrrhizin and some analogues induce growth of primary cultured adult rat hepatoeytes via epidermal growth factor receptors. European Journal of Pharmacology, 2001, 431:151-161). However, due to poor water solubility and low bioavailability of glycyrrhetinic acid, and long-term use can easily cause side effects such as hyperaldosteronism, chemical modification and structural modification of it to improve its water solubility and reduce its side effects have become research hotspots at home and abroad. .

In addition, human diseases and crop diseases caused by bacteria or fungi are very common. The current problem of using chemical drugs to control such diseases is mainly the emergence of bacterial or fungal drug resistance. The application of chemical drugs to pesticides will also lead to environmental pollution, Serious problems such as pesticide residues. Glycyrrhetinic acid has anti-Candida albicans (Pellati D, Fiore C, Armanini D, Rassu M, Bertoloni G.In vitro effects of glycyrrhetinic acid on the growth of clinical isolates of Candida albicans.Phytotherapy Research,2009,23(4),572 -574.), Flavobacterium columnare (Schrader KK.Plant natural compounds with antibacterial activity towards common pathogens of pond-cultured channel catfish(Ictalurus punctatus).Toxins,2010,2(7),1676-1689.), Methicillin-resistant Staphylococcus aureus (Long DR, Mead J, Hendricks JM, Hardy ME, Voyich JM. 18β-Glycyrrhetinic acid inhibits methicillin-resistant Staphylococcus aureus survival and attenuates acute virulence. and chemotherapy, 2013, 57(1), 241-247.) and other activities. Glycyrrhetinic acid has strong antibacterial activity and no pollution to the environment, so its application in medicine and pesticide has a good development prospect.

Tobacco mosaic virus infection affects the normal growth of tobacco and the quality of tobacco leaves. Studies have found that some triterpenes and triterpene saponins have strong anti-tobacco mosaic virus activity, and glycyrrhetinic acid has strong anti-animal virus activity, but its anti-tobacco mosaic virus activity has not been reported.

Contents of the invention

The purpose of the present invention is to provide A-ring polyoxidized substituted glycyrrhetinic acid derivatives and their preparation methods and pharmaceutical applications. The present invention modifies the structure of glycyrrhetinic acid, and synthesizes a series of derivatives substituted by oxidation of the A ring. The biological activity test shows that this type of compound has strong hepatoprotective activity, and is effective against Gram-positive bacteria and other bacteria, tobacco, etc. Fungi such as black shank bacteria and tobacco mosaic virus show strong inhibitory activity, and have good application prospects in the fields of medicine and pesticides.

Technical scheme of the present invention: A ring polyoxidation substituted glycyrrhetinic acid derivatives, the general formula of this compound is the following structural formula 1:

Among them: R ₁ is selected from carbonyl or hydroxyl; R ₂ and R ₃ are both hydroxyl or between R ₂ and R ₃ is an epoxy group; R ₄ is selected from -COOH, -COOR ¹ , -CONH ₂ , -CONHR ¹ , - any one of CONR ¹ R ² ; R ¹ and R ² are selected from one of alkyl, substituted or unsubstituted phenyl, substituted or unsubstituted phenylalkyl containing 1-15 carbon atoms, R ¹ and R ² may be the same or different; the substituents in the substituted phenyl and substituted phenylalkyl groups are selected from halogen, hydroxyl, cyano, amino, nitro, mercapto or phenyl containing 1-15 carbons, Acyl, aryl, alkoxy, alkyl; Y is CH ₂ or C=O; the 18-position hydrogen is in α configuration or β configuration; when R ₁ , R ₂ , and R ₃ are hydroxyl, the hydroxyl substitution configuration Each type is α-configuration or β-configuration; when there is an epoxy group between R ₂ and R ₃ , the epoxy group is α-configuration or β-configuration.

The aforementioned A-ring multi-oxidation substituted glycyrrhetinic acid derivatives, when _R2 and _R3 in the general formula 1 is an epoxy group, that is, the compound of formula I:

The compound of formula I is preferably: R ₁ is selected from carbonyl or hydroxyl, and the hydroxyl is in α configuration or β configuration; the epoxy group between R ₂ and R ₃ is in α configuration or β configuration; R ₄ is selected from -COOH , benzyl ester group or benzyl amino group; Y is CH ₂ or C=O; the 18-position hydrogen is in the β configuration.

More preferred compounds of formula I are: Ia. 3-oxo-(1α,2α)-1,2-epoxy-glycyrrhetinic acid; Ib.3-oxo-(1α,2α)-1,2-ring Oxy-11-deoxyglycyrrhetinic acid; Ic.3-Oxo-(1α,2α)-1,2-epoxy-benzyl glycyrrhetinate; Id.3-Oxo-(1α,2α)-1, 2-Epoxy-11-deoxyglycyrrhetinic acid benzyl ester; Ⅰe.(1α,2α)-1,2-epoxy-glycyrrhetinic acid; Ⅰf.3α-hydroxy-(1α,2α)-1,2-cyclo Oxygen-11-oxo-oleanane-12-ene-30-carboxylic acid; Ⅰg.(1α,2α)-1,2-epoxy-11-deoxyglycyrrhetinic acid; Ⅰh.3α-hydroxyl-( 1α,2α)-1,2-Epoxy-oleanane-12-ene-30-carboxylic acid; Ii. (1α,2α)-1,2-Epoxy-glycyrrhetinic acid benzyl ester; Ij.3α -Hydroxy-(1α,2α)-1,2-epoxy-11-oxo-oleanane-12-ene-30-carboxylic acid benzyl ester; Ik.(1α,2α)-1,2- Epoxy-11-deoxyglycyrrhetinic acid benzyl ester; Il.3α-Hydroxy-(1α,2α)-1,2-epoxy-oleanane-12-ene-30-carboxylate benzyl ester; Im. 3-Oxo-(1β,2β)-1,2-epoxy-glycyrrhetinic acid; In.3-Oxo-(1β,2β)-1,2-epoxy-11-deoxyglycyrrhetinic acid; Io .3-oxo-(1β,2β)-1,2-epoxy-benzyl glycyrrhetinate; Ip.3-oxo-(1β,2β)-1,2-epoxy-11-deoxyglycyrrhetinic acid Benzyl ester; Ⅰq.(1β,2β)-1,2-epoxy-glycyrrhetinic acid; Ⅰr.3α-hydroxy-(1β,2β)-1,2-epoxy-11-oxo-olean Alk-12-ene-30-carboxylic acid; Is.(1β,2β)-1,2-epoxy-11-deoxyglycyrrhetinic acid; It.3α-hydroxy-(1β,2β)-1,2-cyclo Oxy-oleanane-12-ene-30-carboxylic acid; Iu. (1β,2β)-1,2-epoxy-benzyl glycyrrhetinate; Iv.3α-Hydroxy-(1β,2β)-1 , Benzyl 2-epoxy-11-oxo-oleanane-12-ene-30-carboxylate; Iw. (1β,2β)-1,2-epoxy-11-deoxyglycyrrhetinic acid benzyl Esters; Ix. Benzyl 3α-hydroxy-(1β,2β)-1,2-epoxy-oleanane-12-ene-30-carboxylate. Its structural formula is as follows respectively:

The compound of formula I is further preferably: Ib.3-oxo-(1α,2α)-1,2-epoxy-11-deoxyglycyrrhetinic acid; Ic.3-oxo-(1α,2α)-1,2 -Epoxy-benzyl glycyrrhetinate; Ⅰg.(1α,2α)-1,2-Epoxy-11-deoxyglycyrrhetinic acid; Ⅰh.3α-Hydroxy-(1α,2α)-1,2-epoxy -Oleanane-12-ene-30-carboxylic acid; Ik.(1α,2α)-1,2-Epoxy-11-deoxyglycyrrhetinic acid benzyl ester; Il.3α-Hydroxy-(1α,2α) -1,2-Epoxy-oleanane-12-ene-30-carboxylic acid benzyl ester.

The above-mentioned polyoxygenated glycyrrhetinic acid derivatives of the A ring, when R ₁ , R ₂ , and R ₃ in the general formula 1 are all hydroxyl groups, are the compounds of the formula II:

The compound of formula II is preferably: each of the three hydroxyl groups is in α-configuration or β-configuration; R ₄ is selected from -COOH or -COOR ¹ ; R ¹ is selected from alkyl groups containing 1-4 carbon atoms, substituted or unsubstituted One of the phenylalkyl groups, the substituents in the substituted phenylalkyl group are selected from halogen or nitro; Y is CH ₂ or C=O; the hydrogen at position 18 is α-configuration or β-configuration.

More preferred compounds of formula II are: IIa.1α,2α,3α-trihydroxy-11-oxo-oleanane-12-ene-30-carboxylic acid; IIb.1α,2β-dihydroxy-11-oxo Generation-oleanane-12-ene-30-carboxylic acid; Ⅱc.lβ,2α,3α-trihydroxy-11-oxo-oleanane-12-ene-30-carboxylic acid; Ⅱd.lβ, 2β,3α-Trihydroxy-11-oxo-oleanane-12-ene-30-carboxylic acid; IIe.1β,2β-Dihydroxy-glycyrrhetinic acid; IIf.lβ,2α-Dihydroxy-glycyrrhetin acid; Ⅱg.lα,2β,3α-trihydroxy-glycyrrhetinic acid; Ⅱh.1α,2α-dihydroxy-glycyrrhetinic acid; Ⅱi.lα,2α,3α-trihydroxy-oleanane-12-ene- 30-carboxylic acid; Ⅱj.lα,2β,3α-trihydroxy-oleanane-12-ene-30-carboxylic acid; Ⅱk.lβ,2β,3α-trihydroxy-oleanane-12-ene- 30-carboxylic acid; Ⅱl.lβ,2α,3α-trihydroxy-oleanane-12-ene-30-carboxylic acid; Ⅱm.lβ,2β-dihydroxy-11-deoxyglycyrrhetinic acid; Ⅱn.lα, 2β-dihydroxy-11-deoxyglycyrrhetinic acid; IIo.lβ,2α-dihydroxy-11-deoxyglycyrrhetinic acid; IIp.1α,2α-dihydroxy-11-deoxyglycyrrhetinic acid; can also be preferably: R _1. R ₂ , R ₃ are hydroxyl groups in α, β, and β configurations, and Y is C=O; or R ₁ , R ₂ , and R ₃ are hydroxyl groups in α, β, and α configurations, and Y is CH ₂ ; R ₄ is -COOR ¹ , and R ¹ is selected from linear alkyl or isopropyl groups containing 1-4 carbon atoms; the hydrogen at position 18 is α-configuration or β-configuration. The structural formulas of compounds IIa～IIp are as follows:

Compounds of formula II are further preferably: IIa.1α,2α,3α-trihydroxy-11-oxo-oleanane-12-ene-30-carboxylic acid; IIb.1α,2β-dihydroxy-11-oxo -Oleanane-12-ene-30-carboxylic acid; IIg.lα,2β,3α-Trihydroxy-glycyrrhetinic acid; IIj.lα,2β,3α-Trihydroxy-oleanane-12-ene- 30-Carboxylic acid; II m.lβ,2β-dihydroxy-11-deoxyglycyrrhetinic acid.

The preparation method of the aforementioned compound of formula I is as follows: Glycyrrhetinic acid is dissolved in N,N-dimethylformamide (DMF), and potassium carbonate powder is added, and benzyl bromide and carboxyl group are esterified to obtain benzyl compound G-1; Dissolve G-1 in absolute ethanol, add zinc powder and drop in hydrochloric acid, the 11-position carbonyl group is reduced by zinc powder to obtain Gb; G-1 and Gb are respectively dissolved in acetone, and oxidized by Jones reagent to obtain compounds G-2 and Gc; G-2 and Gc were respectively dissolved in acetic anhydride and oxidized by selenium dioxide to obtain α, β-unsaturated ketone _compounds G-3 and Gd; _O2 is oxidized to epoxy compounds Ic, Id and isomers Io, Ip; Ic, Id, Io, Ip are dissolved in methanol and reduced by sodium borohydride to obtain Ii, Ij, Ik, Il, Iu, Iv, Iw, Ix Benzyl ester compounds Ic, Id, Io, Ip, Ii, Ij, Ik, Il, Iu, Iv, Iw, and Ix were dissolved in methanol respectively, and the benzyl group was removed by palladium carbon and hydrogen catalytic hydrogenation to obtain the formula I. R ₄ is -COOH compounds Ia, Ib, Im, In, Ie, If,

Ig, Ih, Iq, Ir, Is, It; the reaction scheme is as follows:

The preparation method of another part of the compound of formula I is: using the compound whose R ₄ is -COOH in formula I as raw material, dissolved in N,N-dimethylformamide (DMF), and reacting with the corresponding haloalkane R ¹ under the action of potassium carbonate X reacts to obtain the corresponding ester compound, that is, the compound in which R ₄ is -COOR ¹ in formula I; the raw material is dissolved in N,N-dimethylformamide (DMF), and in N,N'-dicyclohexylcarbodiethylene Under the action of amine (DCC) and 4-dimethylaminopyridine (DMAP), react with the corresponding amines R ¹ NH ₂ , R ¹ R ² NH to obtain the corresponding amide compounds, that is, in formula I, R ₄ is -CONHR ¹ , The compound of -CONR ¹ R ² ; the raw material is dissolved with dichloromethane (DCM), first reacts with oxalyl chloride (COCl) ₂ to generate an acyl chloride compound, then the acyl chloride compound is dissolved with toluene, and added ammonia water is reacted to obtain R in the formula I _as- The compound of _CONH ; Its reaction scheme is as follows:

The preparation method of the aforementioned compound of formula II is as follows: using compounds Ii, Ij, Iu, Iv, Ik, Il, Iw, and Ix as raw materials respectively, dissolving them in acetone, adding distilled water, and opening the ring with perchloric acid to obtain the corresponding benzyl-substituted Ring-opening compound, then the ring-opening compound is dissolved in methanol, through catalytic hydrogenation of palladium carbon and hydrogen to obtain the compound of _R in the formula II for -COOH; its reaction scheme is as follows:

The preparation method of another part of the compound of formula II is as follows: using compounds IIa~IIp as raw materials, dissolving them in N,N-dimethylformamide (DMF), and reacting with the corresponding haloalkane R ¹ X under the action of potassium carbonate to obtain the corresponding ester Compounds, that is, compounds in which R ₄ is -COOR ¹ in formula II; raw materials are dissolved in N,N-dimethylformamide (DMF), and dissolved in N,N'-dicyclohexylcarbodiimide (DCC) and 4 -Dimethylaminopyridine (DMAP) reacts with the corresponding amines R ¹ NH ₂ , R ¹ R ² NH to obtain the corresponding amide compounds, that is, in formula II, R ₄ is -CONHR ¹ , -CONR ¹ R ² Compound; raw material dissolves with dichloromethane (DCM), reacts with oxalyl chloride to generate acid chloride compound, then dissolves acid chloride compound with toluene, adds ammonia water and reacts to obtain formula II in R ₄ is-CONH ₂ compound; its reaction scheme is as follows:

The above-mentioned A-ring polyoxygenated glycyrrhetinic acid derivatives (that is, compounds of the general formula 1) and their pharmaceutically acceptable salts have applications in the preparation of medical drugs for treating liver damage diseases. The compound of general formula 1 is reacted with basic substances such as sodium hydroxide, potassium hydroxide, calcium hydroxide, etc. to form pharmaceutically acceptable salts, such as sodium salts, potassium salts, or calcium salts, etc., and these salts can be used as liver-protecting drugs.

The above-mentioned A-ring polyoxygenated glycyrrhetinic acid derivatives (that is, compounds of the general formula 1) and their pharmaceutically acceptable salts have applications in the preparation of antibacterial or anti-tobacco mosaic virus drugs.

In the aforementioned applications, the antibacterial drugs are antibacterial or antifungal drugs. Described antibacterial medicine is preferably the agricultural medicine that prevents and treats the crop disease that is caused by Gram-positive bacteria or the medical medicine that treats the infectious disease that is caused by Gram-positive bacteria; Described antifungal medicine is preferably the prevention and treatment caused by Agricultural medicine for crop diseases caused by tobacco blackleg fungus.

In the aforementioned applications, the anti-tobacco mosaic virus drug is an agricultural drug for preventing and treating tobacco diseases caused by tobacco mosaic virus.

In the aforementioned applications, the agricultural drug also contains commonly used pesticide adjuvants; the medical drug also contains commonly used pharmaceutical carriers or excipients. Agricultural drugs can be powders, suspensions, aerosols, granules or emulsions; medical drugs can be solid oral preparations (such as tablets, capsules, dripping pills, etc.), liquid oral preparations or injections.

The following are the biological activity screening and test results of the inventors of the general formula 1 compound and its pharmaceutically acceptable salts:

A. Protective effect on carbon tetrachloride-induced liver cell injury

1. The toxicity of the glycyrrhetinic acid derivatives to human liver cell line HL-7702 cells

Test compounds: Glycyrrhetinic acid (GA), Ia～Ix, Ia-Na, Ib-Na, Ie-Na, If-Na, Ig-Na, Ih-Na, Im-Na, In-Na, Iq- Na, Ir-Na, Is-Na, It-Na, IIa～IIp, IIa-Na～IIp-Na and ester and amide derivatives, the structures of ester and amide derivatives are as follows (wherein G-6 -4 represents the ester derivatives of Ⅱg, G-6-2, G-8, GF-72, GNO-7 represent the ester derivatives of Ⅱb, GF-73 represents the ester derivatives of Ⅱa, GFO-74 represents Ester derivatives of Ii and Ie, GNO-h and GF-H3 represent ester derivatives of Iin, GFO-715 represent ester derivatives of Ic and Ia, GO-7 represent ester derivatives of Ij and If, GNP-7 represents the ester derivatives of Ⅱd, GN-71 represents the amide derivatives of Ⅱe, GN-72 represents the amide derivatives of Ⅱg, G-g-c, G-i, GF-H4, GLP-h represent the ester derivatives of Ⅱj GN-g1 represents amide derivatives of IIm, GF-H2 represents ester derivatives of IIo, GFN-H15 represents amide derivatives of Is, GFI-H3 represents ester derivatives of Id and Ib, GLO- h represents the ester derivatives of Il and If, and GO-h represents the amide derivatives of In):

Main instruments, medicines and reagents: Fluorescence inverted phase-contrast microscope is a product of Nikon, Japan (model: Ti-U); Model 550 microplate reader is a product of American BIO~RAD; 96-well culture plate is a product of Corning Costar, USA; CO2 incubator It is a product of Cellstar Company in the United States; normal human liver cell line HL-7702 cells are provided by the Shanghai Cell Bank of the Chinese Academy of Sciences; DMEM medium, calf serum, Gibco Company; bicyclol, content 99.7% (HPLC), China Institute for the Control of Pharmaceutical and Biological Products ; silibinin, content 98% (HPLC), Xi'an Feida Biotechnology Co., Ltd.; 100U/ml penicillin, thiazolium blue (MTT), dimethyl sulfoxide (DMSO), Sigma company.

Method: Normal liver cell lines were inoculated in 96-well plates at 5×10 ⁴ /m, with a volume of 100 μL per well. 100 μL of the culture medium solution of the test compound, one concentration per 3 wells, placed in a 37°C carbon dioxide incubator and cultured for 48 hours, then added 20 μL of 5 mg/ml MTT to each well, continued in the incubator for 4 hours, sucked out the culture supernatant, and Add 100 μL of DMSO to the wells, shake to dissolve the crystals completely, and measure the absorbance value of each well at 570 nm on an enzyme-linked immunosorbent detector. Calculate the inhibition rate, and then use the spss software for statistical analysis and calculation of the half inhibitory concentration (IC ₅₀ ). Each experiment was repeated three times.

Results: The results of the cytotoxicity test are shown in Table 1. The results showed that most of the compounds with general formula 1 had IC ₅₀ values greater than 100 μmolL ^-1 , indicating that these compounds were less toxic to HL-7702 cells.

2. The protective effect of the glycyrrhetinic acid derivative on the injury of human hepatocyte cell line HL-7702 induced by carbon tetrachloride (CCl ₄ ).

Test compound is identical with human liver cell line HL-7702 cytotoxicity test, main drug and reagent: bicyclol, content 99.7% (HPLC), Chinese institute for the control of pharmaceutical and biological products; silibinin, content 98% (HPLC), Xi'an Feida Biotechnology Co., Ltd.; thiazolyl blue (MTT), Sigma; dimethyl sulfoxide, Sigma; carbon tetrachloride, analytically pure, Chongqing Chuanjiang Chemical Co., Ltd.; DMEM medium, calf serum, Gibco; normal Human liver HL-7702 cells were provided by the Shanghai Cell Bank of the Chinese Academy of Sciences.

Methods: Using MTT method, HL-7702 cells were cultured in RPMI1640 medium containing 10% fetal bovine serum, and cultured statically at 37°C and 5% CO ₂ saturated humidity. Cells in the logarithmic growth phase were inoculated on a 96-well plate at a density of 5×10 ⁴ /ml per well, with a volume of 100 μL per well, cultured and grouped after the cells adhered to the wall: blank control group: untreated normal HL-7702 cells; CCl ₄ Model group: containing 60% saturated _CCl4 culture medium for 6h; positive drug (silibinin and bicyclol) control group; test compound group: set 6 concentration gradients in the concentration range lower than ₅₀ % IC50, the After the cells were cultured for 12 hours in the medium containing the compounds at various concentrations, they were replaced with medium containing 60% saturated CCl ₄ and cultured for 6 hours. Add 10 μL of MTT to each well of each group and continue to culture for 4 hours, discard the supernatant, add 100 μL dimethyl sulfoxide to fully dissolve the purple crystals, measure the absorbance value of each well at 570 nm, calculate the protection rate, and then use spss software for statistical analysis and analysis. Calculation of the half effective concentration (EC ₅₀ ). Each experiment was repeated three times.

Results: The test results of in vitro hepatoprotective activity are shown in Table 1. This kind of glycyrrhetinic acid derivatives has high protective activity against _CCl4 -induced HL-7702 cell damage, and the activity of most compounds is significantly stronger than that of glycyrrhetinic acid. Among them, compound G -8, G-6-2, GF-H3, GNO-7, GNP-7, Ic, IIa-Na, IIb, IIb-Na, IIj, IIn-Na, and IIp-Na were similar to the positive control drug. The ratio TI value (therapeutic index) of IC ₅₀ and EC ₅₀ reflects the safety of the drug. It can be seen from Table 1 that compounds Ⅰh, Ⅰc, Ⅱa, Ⅱa-Na, Ⅱb, Ⅱb-Na, Ⅱd, Ⅱd-Na, Ⅱj, G -6-2, G-8, GF-72, GFO-74, Gi, GLO-h, GN-g1, GNO-7, GNP-7, GO-h and the positive control drugs had good safety.

Table 1 The toxicity of glycyrrhetinic acid derivatives to HL-7702 cells and the protective effect of CCl ₄ on HL-7702 cell damage

compound IC ₅₀ (μmolL ^-1 ) EC ₅₀ (μmolL ^-1 ) Ti bicyclol >200 53.86 >3.37 Silybin >200 47.14 >4.2 Im >200 138.52 >1.45 Im-Na >200 114.38 1.75 Io >200 >150 >1.33 IP 166.93 142.18 1.17 Ix 190.34 100.84 1.89 it 102.71 69.34 1.48 It-Na 113.28 78.02 1.45 I u >200 >150 >1.33 IV 118.21 89.06 1.33 Iw 148.25 111.18 1.33 Ig 173.52 102.35 1.70 Ig-Na 157,39 87.03 1.80 Ih >200 69.21 >2.89 Ih-Na 157.37 75.73 2.07 Ia >200 129.31 >1.55 Ia-Na >200 97.39 2.05 Ib 158.13 90.77 1.74 Ib-Na 131.12 93.87 1.39 Ic 173.52 56.00 3.10 Id >200 >150 >1.33 I e 147.36 100.79 1.46 Ie-Na 100.74 89.07 1.12 If >200 129.37 >1.55 If-Na 176.39 100.84 1.75 In 95.27 78.10 1.22 In-Na 100.37 57.03 1.76 Iq 174.06 137.11 1.27 Iq-Na >200 119.37 1.67 Ii >200 >150 >1.33 Ij 110.65 129.53 0.85 Ik 174.71 >150 <1.16 Il 76.40 145.07 0.53 Ir 99.28 67.43 1.47 Ir-Na 89.03 60.98 1.46 Is >200 >150 >1.27 Is-Na 156.38 139.03 1.12 IIa 168.96 61.31 2.76 IIa-Na 154.98 54.80 2.80 IId 198.99 57.38 3.49 IId-Na 178.34 60.93 2.92 IIb 150.75 51.62 2.92 IIb-Na 147.54 50.57 2.91 IIc 99.31 57.03 1.74 IIc-Na 87.39 58.09 1.50 IIe 127.23 84.02 1.51 IIe-Na 132.89 76.58 1.73 IIf 28.30 65.90 0.43 IIf-Na 32.78 70.47 0.46 II g 75.75 71.29 1.06 IIg-Na 78.29 65.98 1.18 IIi 126.48 75.42 1.68 IIi-Na 94.37 77.30 1.22 IIh 172.37 128.02 1.35 IIh-Na 153.22 130.37 1.17 IIk 127.87 71.71 1.78 IIk-Na 139.03 78.88 1.76 IIj 116.58 52.55 2.22 IIj-Na 99.37 63.25 1.57 IIn 27.35 59.65 0.46 IIn-Na 89.03 53.78 1.65 IIl 138.47 68.02 2.03 IIl-Na 120.77 59.82 2.01 IIm 101.28 59.99 1.69 IIm-Na 120.10 65.02 1.85 IIp 87.03 57.84 1.50 IIp-Na 88.76 54.00 1.64 IIo 32.43 58.99 0.55 IIo-Na 77.35 55.28 1.39 G-6-2 146.70 57.38 2.56 G-6-4 123.11 85.42 1.44 G-8 157.01 49.29 3.19 GA 169.17 125.52 1.35 GF-72 130.34 58.99 2.21 GF-73 133.98 >150 >0.89 GF-H2 73.16 57.85 1.26 GF-H3 99.29 56.00 1.77 GF-H4 130.92 69.90 1.87 GFI-H3 161.15 80.08 2.01 GFN-H15 97.96 >150 <0.64 GFO-715 >200 120.21 >1.66 GFO-74 >200 65.67 >3.07 Ggc 25.30 78.71 0.32 Gi 150.42 60.29 2.49 Glo-h 193.87 60.18 3.22 GLP-h 102.30 71.51 1.43 GN-71 157.35 84.29 1.87 GN-72 167.93 91.72 1.83 GN-g1 128.43 53.53 2.40 GNO-7 132.62 56.64 2.34 GNO-h 131.43 92.38 1.42 GNP-7 154.94 56.04 2.76 GO-7 >200 129.53 >1.55 GO-h 155.87 61.27 2.54 3. The protective effect of the glycyrrhetinic acid derivatives on _CCl4- induced acute liver injury in mice Test compounds: Ⅱb, Ⅱb-Na, Ⅰc, Ⅱd, Ⅱd-Na, Ⅱj, GN-g1, GNP-7, G -8, G-6-2, Gi. All the above compounds were mixed with 0.5% CMC-Na to prepare the required concentration suspension before the experiment. Main drugs and reagents: Silymarin, 90% (HPLC), Xi'an Feida Biotechnology Co., Ltd.; Bicyclol Tablets, Beijing Union Pharmaceutical Factory, batch number: 100602. Carboxymethylcellulose sodium (CMC-Na), analytically pure, Tianjin Kemiou Chemical Reagent Co., Ltd. Animals: Healthy KM mice, half male and half female, weighing 20-25 g, provided by Chongqing Tengxin Bill Experimental Animal Sales Co., Ltd., license number: SCXK (Chongqing) 20120006. Male and female mice were separated into cages, with 9 mice in each cage, and were raised in a clean animal breeding room with sufficient light and good ventilation, with a room temperature of 18-25°C and a relative humidity of 50-70%.

Methods: 168 Kunming mice were randomly divided into 7 groups: normal control group, model control group, silymarin positive control group (50mg/kg), bicyclol (50mg/kg) positive control group and test subjects. Compound high (100mg/kg), middle (50mg/kg), low (25mg/kg) dose groups. 6 rats in each group were administered by intragastric administration. The drug was dissolved in 0.5% CMC-Na, and the administration volume was 25ml/kg. The normal control group and the model group were given an equal volume of 0.5% CMC-Na. 2 hours after the last administration, except for the normal control group, all were injected intraperitoneally with 0.1% carbon tetrachloride olive oil solution 10ml/kg, and 24 hours later, the eyeballs were removed to collect blood (16 hours before blood collection), and the serum obtained by centrifugation was used as ALT (alanine aminotransferase), AST (aspartate aminotransferase) and ALP (alkaline phosphatase) detection, a small piece of liver from the same part of the left lobe of the mouse liver was fixed in neutral 10% formalin phosphate solution, and embedded in paraffin Slices were stained with HE, and the histopathological changes of the liver were observed under a light microscope. The data were statistically processed with spss software.

Results: The detection results of ALT, AST and ALP in mouse serum are shown in Table 2, and the observations of liver histopathological changes are shown in Figures 1 to 10. It can be seen from Table 2 that compared with the normal control group, the serum ALT, AST and ALP activities of the mice in the model group CCl ₄ induced acute chemical liver injury were significantly increased (P<0.01 ), suggesting that the liver injury model was established successfully. Compared with the model group (p<0.05), the tested drugs could reduce the activities of the three enzymes in a dose-dependent manner. The low-dose compound G-8 group was significantly better than the two positive drugs in reducing the activity of ALT, while the middle-dose and high-dose groups could make the activities of the three enzymes tend to normal. Histopathological observation of the mouse liver showed that in the normal control group, the liver tissue structure was normal, the structure of the hepatic lobule was intact, the arrangement of the liver cord was regular, there was no abnormality in the hepatic sinusoidal area and the portal area, no inflammatory cell infiltration, and the morphology of the liver cells was normal (Figure 1 1, 2); in the CCl ₄ acute liver injury model group, hepatocyte swelling, loose cytoplasm, irregular cell shape, unclear cell boundaries, fatty degeneration, obvious punctate and focal necrosis, portal area accompanied by neutral Infiltration of inflammatory cells such as granulocytes and lymphocytes (3 and 4 in Figure 1); compared with the model group, the silymarin-positive control group had significantly less liver tissue damage, manifested as less punctate and focal necrosis, and less inflammatory cell infiltration in the portal area , the cell morphology is more regular, and the fatty degeneration is reduced (5, 6 in Figure 1); the changes of the test compound on the liver histopathology of mice are shown in Figure 2 to Figure 10, and it can be seen from the figure that these compounds can reduce the damage of liver cells , and presented a dose-dependent manner. Among them, compound G-8 has the best effect, and its low-dose group can significantly reduce liver cell damage, showing that the morphology and structure of liver cells are relatively complete, the cell boundary is clear, only a small amount of cell necrosis, and less inflammatory cell infiltration (Fig. 2 of 1, 2); G-8 middle-dose group has relatively complete liver tissue structure, regular arrangement of liver cords, and less inflammatory cells (Fig. 2, 3, 4); G-8 high-dose group has normal cell structure and Clear, almost no inflammatory cell infiltration, no cell necrosis (Figure 2, 5, 6). The above results showed that the protective effect of compound G-8 on CCl ₄ -induced acute liver injury in mice was better than that of positive control drugs silymarin and bifendate.

Table 2 Protective effect of glycyrrhetinic acid derivatives on _CCl4 -induced acute liver injury in mice

group ALT(U/L) AST(U/L) ALP(U/L) normal group 22.17±2.01 89.17±6.21 163.83±5.51 CCl ₄ model group 227.17±5.48 ^# 183.00±8.23 ^# 232.17±2.45 ^# IIb (25mg/kg) 163.30±3,87 ^** 201.48±1.90 ^* 221.47±4.89 IIb (50mg/kg) 148.35±5.29 ^** 199.93±4.76 195.38±5.02 ^** IIb (100mg/kg) 87.38±2.88 ^** 147.87±2.45 ^** 168.04±2.77 ^** Ⅱb-Na(25mg/kg) 183.50±3.45 ^* 224.17±4.57 191.00±2.16 ^* Ⅱb-Na(50mg/kg) 84.25±1.35 ^** 137.25±2.46 ^** 165.50±1.89 ^** Ⅱb-Na(100mg/kg) 58.80±1.24 ^** 119.00±3.00 ^** 157.60±1.88 ^** IId (25mg/kg) 220.87±1.45 189.37±3.48 230.28±4.87 IId (50mg/kg) 201.55±2.00 ^* 162.59±3.26 ^* 207.35±4.55 ^* IId (100mg/kg) 158.74±2.53 ^** 120.38±3.70 ^** 179.37±3.98 ^** Ⅱd-Na(25mg/kg) 201.35±3.44 169.38±5.56 223.72±1.46 Ⅱd-Na(50mg/kg) 174.28±3.89 ^* 157.88±3.22 ^* 208.46±2.18 ^* Ⅱd-Na(100mg/kg) 133.79±2.60 ^** 136.07±4.75 ^* 185.90±1.78 ^* G-8 (25mg/kg) 48.50±2.38 ^** 113.67±3.51 ^** 212.33±1.80 G-8 (50mg/kg) 37.00±2.67 ^** 75.50±5.38 ^** 168.00±2.60 ^** G-8 (100mg/kg) 34.00±1.79 ^** 70.00±3.87 ^** 166.00±1.99 ^** G-6-2 (25mg/kg) 213.00±5.43 175.67±4.33 231.00±4.29 G-6-2 (50mg/kg) 215.40±4.21 151.20±4.87 ^* 219.20±2.68 G-6-2 (100mg/kg) 149.17±3.27 ^** 105.17±3.88 ^** 173.00±2.90 ^**

IIj (25mg/kg) 196.50±4.19 183.00±5.29 195.83±3.45 ^* IIj (50mg/kg) 112.33±4.28 ^** 95.50±6.48 ^** 207.83±2.87 IIj (100mg/kg) 109.67±3.21 ^** 104.83±3.25 ^** 183.00±1.70 ^* G-i (25mg/kg) 188.37±2.87 ^* 150.29±4.16 ^* 216.78±4.24 G-i (50mg/kg) 147.38±4.27 ^** 133.28±3.56 ^** 199.35±2.50 ^* G-i (100mg/kg) 127.89±3.22 ^** 99.72±4.51 ^** 188.06±2.48 ^** Ic (25mg/kg) 199.03±5.66 ^* 168.39±4.28 228.68±3.11 Ic(50mg/kg) 147.91±3.27 ^** 143.88±3.90 ^** 198.18±2.33 Ic(100mg/kg) 101.37±3.24 ^** 92.84±2.44 ^** 179.48±2.17 ^** GN-g1 (25mg/kg) 189.33±4.38 ^* 205.50±4.88 204.38±1.28 ^* GN-g1 (50mg/kg) 163.20±4.19 ^** 184.84±5.11 185.89±2.01 ^** GN-g1 (100mg/kg) 138.77±3.55 ^** 150.32±4.10 ^* 184.20±2.03 ^** GNP-7 (25mg/kg) 190.68±4.06 ^* 210.31±5.90 201.42±2.88 ^* GNP-7 (50mg/kg) 105.76±6.01 ^** 90.43±3.28 ^** 188.91±2.00 ^* GNP-7 (100mg/kg) 87.99±3.77 ^** 88.00±2.60 ^** 156.33±1.58 ^** Silymarin (50mg/kg) 60.50±4.17 ^** 61.50±3.01 ^** 136.17±1.30 ^** Bifendate (50mg/kg) 81.40±4.21 ^** 151.80±4.05 ^** 186.40±2.00 ^**

Compared with normal group, #p<0.01; compared with CCl ₄ model group, *p<0.05, **p<0.01.

In summary, the A-ring polyoxidized substituted glycyrrhetinic acid derivatives provided by the present invention have a significant protective effect on the acute liver injury in mice caused by CCl ₄ , and are expected to be used as hepatoprotective drugs.

B. Antibacterial effect

4. The antibacterial activity screening experiment of the glycyrrhetinic acid derivatives

Test compounds: Glycyrrhetinic acid (GA), Ia～Ix, Ia-Na, Ib-Na, Ie-Na, If-Na, Ig-Na, Ih-Na, Im-Na, In-Na, Iq- Na, Ⅰr-Na, Ⅰs-Na, It-Na, Ⅱa～Ⅱp, Ⅱa-Na～Ⅱp-Na and ester and amide derivatives in the liver protection activity test, and some ester and amide derivatives are added object, its structure is as follows:

Experimental strain:

Gram-positive bacteria: Staphylococcus aureus, Bacillus subtilis, Streptomyces scabiei;

Gram-negative bacteria: Escherichia coli, Ralstonia solanacearum.

Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Ralstonia solanacearum were provided by the Microbiology Laboratory of the College of Life Sciences, Guizhou University; scabiei) was purchased from Beijing Beina Chuanglian Biotechnology Research Institute.

Main instruments and reagents: -80 ℃ ultra-low temperature refrigerator (ULT1386, American Thermo Company); automatic triple pure water distiller (SZ-97, Shanghai Yarong Biochemical Instrument Factory); ultra-clean bench (SW-CJ-1F, Suzhou Purification Equipment Co., Ltd.); magnetic heating stirrer (C-MAG HS4, German IKA company); constant temperature shaking incubator (HZQ-F160, Jiangsu Taicang Experimental Equipment Factory); electronic balance (SL-114, American DENVER company); Model micropipette (Germany Eppendorf Company); 96-well culture plate is the product of Corning Costar Company in the United States; Model l550 microplate reader is the product of American BIO-RAD Company; tryptone and yeast extract are purchased from British Oxoid Company; Sulfoxide, penicillin and streptomycin are all products of sigma company.

Preparation of bacterial culture medium: (1) Preparation of beef extract-peptone medium: Weigh 3.0g of beef extract, 10.0g of peptone, and 5.0g of sodium chloride, add 50mL of distilled water, heat and stir continuously to make it completely melted, and adjust The pH value of the solution is 7.2, and finally make up distilled water to 1000mL, aliquot, stopper and autoclave. (2) Preparation of potato culture medium (PDA medium): take 200 g of peeled and washed potatoes, cut into small pieces, put in 1000 mL of distilled water and boil for 30 minutes, then filter with 4 layers of gauze to obtain the filtrate, add glucose to the filtrate 20g, stir to dissolve, make up the missing water to 1000mL, aliquot, stopper and autoclave. The medium prepared by the above method is all liquid medium, and 15%-20% agar is added to obtain the corresponding solid medium.

Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Ralstonia solanacearum were cultured in beef extract peptone medium with an optimum temperature of 37°C; Streptomyces scab was cultured in potato medium with an optimum temperature of 28°C.

method:

Preparation of bacterial suspension: Take the spare bacterial strains of the above 6 kinds of bacteria to be tested, use an inoculation loop to inoculate a little bacterial lawn on the corresponding solid medium slope, and activate at the optimum temperature. Then put the activated strain in 7mL liquid culture medium in a loop, and cultivate it in a constant temperature shaker until the culture medium is turbid to make a bacterial suspension.

Firstly, the antibacterial activity of the compounds was screened by 96 microwell turbidity method. A test compound group (100 μmol/L), a positive drug control group (streptomycin sulfate and ampicillin concentrations both 100 μmol/L) and a blank control group were established. The compounds were all dissolved in DMSO, and the concentration of DMSO was not more than 1%. Adjust the concentration of the bacterial suspension to 10 ⁶ CFU/ml, add 135 μL/well into a 96-well plate, then add 15 μL of the test compound and positive drug solution to each well, so that the final concentration of each compound is 100 μmol/L, and the blank control group is Replace with the same volume of culture medium, set 3 duplicate wells for each compound, measure the absorbance value A ₁ of each well at 600nm with a microplate reader, and then fix the 96-well plate on a constant temperature oscillator with an object support level Medium, with 200rpm rotation speed, cultivated at the optimum temperature. When the bacterial growth reaches the logarithmic growth phase, stop the culture, blow the bacterial solution in each well evenly under aseptic operation, measure the absorbance value A ₂ again at 600nm with a microplate reader, and calculate the difference ΔA between the two absorbance values before and after, according to the following formula Find the inhibition rate:

Inhibition rate = (blank group ΔA - drug group ΔA) * 100% / blank group ΔA

Each experiment was repeated three times, and the average inhibition rate was calculated.

Results: See Tables 3, 4, 5, 6 and 7 for the results of the inhibitory activity of the tested compounds against 5 kinds of bacteria. It can be seen from the table that this type of compound has weak inhibitory activity to Escherichia coli and tobacco Ralstonia solanacearum, and most of the compounds have significant inhibitory activity to Bacillus subtilis, Staphylococcus aureus and Streptomyces scabies, the inhibitory activity of some compounds and Positive control drugs are similar, such as compounds GF-H4, Ⅱj, and GF-72 all have strong inhibitory effects on Staphylococcus aureus and Streptomyces scab. The above results show that the compounds have no or weak inhibitory activity against Gram-negative bacteria, but have strong inhibitory activity against Gram-positive bacteria.

Table 3 The inhibitory effect of glycyrrhetinic acid derivatives on Escherichia coli (concentration: 100 μ M)

compound Inhibition rate(%) compound Inhibition rate(%) GA 6.25±2.29 IIm 0.05±2.14 Ia 0.56±1.04 IIk 3.28±1.06 Ib 0.10±2.32 IIj 1.58±0.54 Ic 0.91±1.17 IIo 2.20±1.74 Id 0.49±1.46 IIb 2.35±1.80 I e 0.25±0.67 IIc 0.67±2.17 If 0.00±1.03 IId 1.00±2.60 Ig 2.25±2.28 IIe 3.46±1.04 Ih 1.83±0.78 IIf 8.67±2.33 Ii 2.00±1.65 IIg 5.57±2.28 Ij 0.00±0.50 IIh 8.80±3.15 Ik 0.05±1.14 IIi 9.03±1.03 Il 2.54±2.34 IIl 9.91±1.76

Im 2.00±2.09 II n 3.80±3.09 In 0.60±1.03 IIp 0.28±0.04 Io 0.58±1.81 IIa-Na 8.30±5.52 IP 3.02±1.42 IIb-Na 4.87±2.78 Iq 0.28±1.37 IIc-Na 2.43±1.56 Ir 3.00±1.57 IId-Na 3.04±2.59 Is 4.11±1.00 IIe-Na 4.80±0.99 it 0.00±1.37 IIf-Na 5.58±3.08 I u 0.58±2.47 IIg-Na 5.80±3.99 IV 1.66±2.57 IIh-Na 6.02±1.13 Iw 1.80±2.79 IIi-Na 10.98±2.49 ^* Ix 0.00±4.88 IIj-Na 7.21±1.48 Ia-Na 5.89±1.94 IIk-Na 6.39±0.84 Ib-Na 5.88±3.00 IIl-Na 6.80±1.78 Ie-Na 6.81±1.44 IIm-Na 9.31±3.79 ^* If-Na 2.55±1.51 IIn-Na 11.38±2.45 ^* Ig-Na 2.88±2.48 IIo-Na 6.28±2.11 Ih-Na 3.70±2.73 IIp-Na 7.80±1.44 Im-Na 4.79±3.85 G-6-2 24.68±2.29 ^** In-Na 4.28±7.08 GF-74 7.92±2.88 Iq-Na 2.66±4.00 GNP-h 5.08±3.80 Ir-Na 0.00±2.37 G-g-c 5.03±2.44 Is-Na 1.68±2.44 GI-H3 0.00±1.21 It-Na 5.37±1.88 Glo-h 8.32±1.40 IIa 5.23±2.24 GO-h 6.77±3.01 GF-H3 0.00±3.05 GLP-h 8.10±2.43 GF-H2 2.32±2.34 GFI-H3 10.44±3.55 ^* G-i 5.70±1.44 GFN-H15 12.84±0.36 ^* GN-g1 6.29±1.70 GF-H4 9.37±1.79 GNO-7 3.66±1.18 GLP-7 27.11±2.88 ^** G-8 8.49±1.00 GF-3 9.36±0.38 G-73 6.80±1.79 GF-73 4.43±2.92 GNO-h 5.53±2.98 GN-72 3.80±1.70 G-6-4 7.80±1.59 GN-71 9.29±2.54 GFO-715 9.09±0.31 GNP-7 4.42±4.77 GFO-74 8.38±1.39 GO-7 8.43±2.45 GF-72 4.88±2.67 streptomycin sulfate 73.91±3.80 ^** blank group 0.00±0.00 Ampicillin 4.60±1.45

Compared with blank control group, ^* P<0.05; ^** P<0.01.

The inhibitory effect of table 4 glycyrrhetinic acid derivatives on Bacillus subtilis (concentration: 100 μ M)

compound Inhibition rate(%) compound Inhibition rate(%) GA 30.17±1.29 ^** IIm 67.45±2.03 ^** Ia 12.67±2.17 ^** IIk 2.32±1.33 Ib 25.38±2.09 ^** IIj 60.17±2.98 ^** Ic 25.17±0.85 ^** IIo 55.20±1.34 ^** Id 31.56±2.11 ^** IIb 42.85±1.88 ^** I e 0.93±1.01 IIc 50.67±4.17 ^** If 20.41±3.57 ^** IId 62.07±2.61 ^**

Ig 9.21±0.82 ^* IIe 73.46±2.54 ^** Ih 53.66±2.08 ^** IIf 58.27±2.13 ^** Ii 35.50±1.57 ^** IIg 65.77±2.08 ^** Ij 9.38±3.55 ^* IIh 78.87±3.25 ^** Ik 19.21±2.18 ^** IIi 19.46±1.00 ^** Il 86.89±1.96 ^** IIl 29.47±1.55 ^** Im 16.63±1.90 IIn 53.62±2.58 ^** In 0.09±2.38 IIp 60.28±3.58 ^** Io 69.21±2.77 ^** IIa-Na 0.00±3.78 IP 63.50±2.73 ^** IIb-Na 26.16±2.05 ^** Iq 23.60±2.57 ^** IIc-Na 42.43±1.47 ^** Ir 32.15±1.09 ^** IId-Na 73.51±2.90 ^** Is 25.41±2.07 ^** IIe-Na 11.32±1.85 ^* it 63.18±4.70 ^** IIf-Na 35.58±3.22 I u 55.71±0.35 ^** IIg-Na 16.29±4.43 ^** IV 43.19±2.16 ^** IIh-Na 62.00±1.35 ^** Iw 5.20±2.01 IIi-Na 60.98±2.10 ^** Ix 0.00±3.17 ^** IIj-Na 58.21±1.36 ^** Ia-Na 25.19±1.44 ^** IIk-Na 76.39±0.48 ^** Ib-Na 15.08±3.30 ^** IIl-Na 76.38±3.08 ^** Ie-Na 56.81±1.48 ^** IIm-Na 49.31±3.00 ^** If-Na 52.57±1.20 ^** IIn-Na 61.28±2.65 ^* Ig-Na 72.81±2.55 ^** IIo-Na 66.21±2.71 ^** Ih-Na 53.20±2.63 ^** IIp-Na 57.40±1.25 ^** Im-Na 64.29±3.88 ^** G-6-2 62.09±0.24 ^** In-Na 64.28±3.01 ^** GF-74 29.93±3.57 ^** Iq-Na 52.86±4.10 ^** GNP-h 12.52±2.68 ^* Ir-Na 50.02±2.87 ^** G-g-c 23.32±2.24 ^** Is-Na 81.68±2.34 ^** GI-H3 11.32±3.30 ^* It-Na 75.28±1.18 ^** Glo-h 78.36±1.26 ^** IIa 65.23±2.66 ^** GO-h 76.77±3.31 ^** GF-H3 80.50±3.37 ^** GLP-h 48.10±2.73 ^** GF-H2 39.36±2.81 ^** GFI-H3 19.44±3.25 ^* G-i 65.82±0.35 ^** GFN-H15 62.54±0.86 ^* GN-g1 46.29±1.60 ^** GF-H4 50.86±3.59 ^** GNO-7 53.76±1.00 ^** GLP-7 60.06±1.38 ^** G-8 88.49±1.47 ^** GF-3 35.89±4.28 ^** GF-73 67.36±3.00 ^** G-73 84.43±2.32 ^** GNO-h 75.53±2.36 ^** GN-72 63.82±3.76 ^** G-6-4 98.81±2.49 ^** GN-71 39.26±2.22 ^** GFO-715 59.09±2.31 ^** GNP-7 74.32±4.50 ^** GFO-74 48.38±1.27 ^** GO-7 68.43±2.22 ^** GF-72 99.63±2.72 ^** streptomycin sulfate 48.56±2.57 ^** blank group 0.00±0.00 Ampicillin 19.05±3.80 ^**

Compared with blank control group, ^* P<0.05; ^** P<0.01.

The inhibitory effect of table 5 glycyrrhetinic acid derivatives on Staphylococcus aureus (100 μ M)

compound Inhibition rate(%) compound Inhibition rate(%) GA 23.97±1.46 ^** IIm 55.87±2.29 ^**

Ia 37.57±2.48 ^** IIk 69.90±2.05 ^** Ib 55.32±4.02 ^** IIj 98.20±2.89 ^** Ic 65.17±1.85 ^** II o 62.34±2.24 ^** Id 36.22±2.68 ^** IIb 49.03±1.37 ^** I e 56.09±3.08 ^** IIc 56.68±4.32 ^** If 22.18±3.33 ^** IId 32.87±2.33 ^** Ig 69.26±5.02 ^** IIe 44.68±2.31 ^** Ih 68.13±2.37 ^** IIf 63.27±3.10 ^** Ii 35.50±1.57 ^** II g 52.51±4.11 ^** Ij 39.16±3.25 ^** IIh 75.36±1.85 ^** Ik 29.01±2.00 ^** IIi 20.46±1.04 ^** Il 96.80±1.16 ^** IIl 36.47±1.68 ^** Im 19.58±1.24 ^** II n 39.42±2.28 ^** In 52.09±2.26 ^** IIp 74.28±3.00 ^** Io 46.28±3.71 ^** IIa-Na 50.45±3.26 ^** IP 53.20±2.03 ^** IIb-Na 46.06±2.87 ^** Iq 33.60±2.17 ^** IIc-Na 49.40±1.73 ^** Ir 70.15±1.03 ^** IId-Na 64.58±1.04 ^** Is 25.41±2.07 ^** IIe-Na 31.35±1.63 ^** it 58.18±3.40 ^** IIf-Na 73.28±2.87 I u 65.71±2.35 ^** IIg-Na 36.99±4.01 ^** IV 37.70±2.33 ^** IIh-Na 69.03±1.66 ^** Iw 65.27±4.18 ^** IIi-Na 37.98±1.70 ^** Ix 68.01±3.00 ^** IIj-Na 78.20±1.05 ^** Ia-Na 35.19±1.00 ^** IIk-Na 53.39±2.08 ^** Ib-Na 35.08±4.87 ^** IIl-Na 70.38±3.15 ^** Ie-Na 50.81±1.40 ^** IIm-Na 41.31±3.06 ^* If-Na 72.53±1.29 ^** IIn-Na 11.28±2.45 ^* Ig-Na 12.81±2.50 ^* IIo-Na 76.26±2.74 ^** Ih-Na 63.20±2.03 ^** IIp-Na 77.40±1.15 ^** Im-Na 44.29±3.28 ^** G-6-2 51.79±2.04 ^** In-Na 69.28±2.01 ^** GF-74 70.71±1.60 ^** Iq-Na 42.76±4.20 ^** GNP-h 10.82±1.91 ^* Ir-Na 62.02±2.47 ^** G-g-c 61.80±3.55 ^** Is-Na 91.68±3.34 ^** GI-H3 43.48±0.46 ^** It-Na 72.20±1.48 ^** Glo-h 88.47±1.16 ^** IIa 27.10±1.33 ^** GO-h 77.24±3.61 ^** GF-H3 85.50±4.69 ^** GLP-h 58.70±2.23 ^** GF-H2 76.57±4.76 ^** GFI-H3 69.14±3.65 ^* G-i 98.85±3.82 ^** GFN-H15 69.34±1.87 ^* GN-g1 56.29±4.65 ^** GF-H4 98.04±2.58 ^** GNO-7 73.36±1.57 ^** GLP-7 30.47±2.70 ^** G-8 89.41±3.47 ^** GF-3 98.32±3.48 ^** GF-73 47.36±3.77 ^** G-73 79.22±3.88 ^** GNO-h 74.53±2.67 ^** GN-72 65.22±3.86 ^** G-6-4 98.10±0.55 ^** GN-71 49.36±2.68 ^** GFO-715 59.09±2.31 ^** GNP-7 31.69±3.89 ^** GFO-74 88.38±1.44 ^** GO-7 78.63±4.00 ^**

GF-72 96.75±2.65 ^** streptomycin sulfate 36.16±3.44 ^** blank group 0.00±0.00 Ampicillin 97.34±2.91 ^**

Compared with blank control group, ^* P<0.05; ^** P<0.01.

The inhibitory effect of table 6 glycyrrhetinic acid derivatives on tobacco Ralstonia solanacearum (100 μ M)

compound Inhibition rate(%) compound Inhibition rate(%) GA 4.79±1.00 IIm 0.00±1.25 Ia 0.00±1.25 IIk 12.93±1.47 ^* Ib 0.36±2.31 IIj 12.31±2.55 ^* Ic 0.00±1.28 II o 0.00±1.04 Id 1.87±1.38 IIb 16.33±3.28 ^** I e 4.25±0.58 IIc 0.00±2.11 If 1.70±1.99 IId 1.34±2.36 Ig 8.25±2.49 IIe 3.58±1.26 Ih 1.03±0.72 IIf 18.07±2.57 ^** Ii 2.88±1.64 IIg 0.06±1.81 Ij 0.00±0.50 IIh 4.93±1.68 Ik 5.05±1.36 IIi 4.03±1.23 Il 5.80±2.00 IIl 0.00±1.71 Im 9.38±0.36 ^* IIn 13.76±3.19 ^** In 6.80±1.21 IIp 3.28±2.04 Io 10.36±1.77 ^* IIa-Na 2.30±1.52 IP 13.02±1.02 ^** IIb-Na 0.87±2.38 Iq 4.28±1.56 IIc-Na 2.54±1.33 Ir 3.66±1.26 IId-Na 0.04±2.19 Is 4.18±1.36 IIe-Na 5.69±2.34 it 7.00±1.06 IIf-Na 0.51±3.03 I u 0.03±2.67 IIg-Na 0.80±3.55 IV 4.66±2.50 IIh-Na 1.02±1.73 Iw 1.00±1.79 IIi-Na 0.58±2.42 ^* Ix 0.50±2.88 IIj-Na 4.21±1.88 Ia-Na 5.59±1.74 IIk-Na 3.39±1.84 Ib-Na 5.28±3.06 IIl-Na 6.20±1.58 Ie-Na 1.81±1.47 IIm-Na 7.31±3.39 If-Na 2.45±1.52 IIn-Na 0.58±2.46 Ig-Na 2.58±2.78 IIo-Na 3.28±2.71 Ih-Na 3.72±2.75 IIp-Na 8.80±1.24 ^* Im-Na 4.29±3.89 G-6-2 2.14±1.59 In-Na 7.28±7.18 GF-74 13.90±1.87 ^** Iq-Na 2.36±6.00 GNP-h 4.64±2.33 Ir-Na 0.70±2.97 G-g-c 2.25±1.72 Is-Na 1.18±2.74 GI-H3 22.70±2.11 ^** It-Na 5.87±1.38 Glo-h 5.32±1.60 IIa 7.11±2.42 GO-h 8.31±3.61 ^* GF-H3 0.46±3.15 GLP-h 2.10±2.44 GF-H2 0.00±1.38 GFI-H3 0.34±3.75 G-i 13.550.38 ^** GFN-H15 12.94±2.36 ^* GN-g1 6.59±1.78 GF-H4 17.25±1.70 ^** GNO-7 7.66±1.48 GLP-7 2.97±0.71

G-8 15.49±1.30 ^** GF-3 5.40±1.30 G-73 24.76±2.08 ^** GF-73 14.73±2.22 ^** GNO-h 15.53±2.68 ^** GN-72 8.80±1.73 G-6-4 14.45±2.68 ^** GN-71 5.18±1.38 GFO-715 9.59±2.31 ^* GNP-7 0.00±1.79 GFO-74 5.38±1.79 GO-7 2.98±3.12 GF-72 0.00±1.00 streptomycin sulfate 76.18±3.01 ^** blank group 0.00±0.00 Ampicillin 9.32±2.49 ^*

Compared with blank control group, ^* P<0.05; ^** P<0.01.

The inhibitory effect of table 7 glycyrrhetinic acid derivatives on streptomyces scab (100 μ M)

compound Inhibition rate(%) compound Inhibition rate(%) GA 35.01±1.88 ^** IIm 97.54±3.47 ^** Ia 26.55±0.48 ^** IIk 2.13±2.87 Ib 61.67±1.27 ^** IIj 98.25±2.04 ^** Ic 69.81±1.24 ^** II o 63.79±2.93 ^** Id 5.36±2.47 IIb 51.09±2.57 ^** I e 68.65±2.33 ^** IIc 76.68±2.32 ^** If 15.84±3.71 ^** IId 62.37±2.53 ^** Ig 63.62±1.48 ^** IIe 38.34±2.29 ^** Ih 86.53±3.12 ^** IIf 65.27±2.10 ^** Ii 61.53±1.82 ^** II g 35.51±1.51 ^** Ij 48.63±2.58 ^** IIh 6.65±2.11 Ik 86.76±2.28 ^** IIi 28.46±1.54 ^** Il 61.47±2.04 ^** IIl 76.47±1.48 ^** Im 56.70±2.60 ^** II n 69.42±2.48 ^** In 53.90±1.40 ^** IIp 54.28±3.70 ^** Io 81.63±1.64 ^** IIa-Na 60.45±3.24 ^** IP 63.76±1.33 ^** IIb-Na 66.06±2.37 ^** Iq 28.53±0.38 ^** IIc-Na 59.40±1.83 ^** Ir 68.67±1.57 ^** IId-Na 74.58±1.74 ^** Is 79.81±1.24 ^** IIe-Na 61.35±1.64 ^** it 4.36±2.44 IIf-Na 76.28±2.37 ^** I u 66.65±2.77 ^** IIg-Na 38.99±2.01 ^** IV 68.84±3.75 ^** IIh-Na 67.03±1.36 ^** Iw 54.62±1.88 ^** IIi-Na 77.98±1.30 ^** Ix 87.53±3.02 ^** IIj-Na 68.20±1.45 ^** Ia-Na 71.56±1.02 ^** IIk-Na 73.39±3.08 ^** Ib-Na 58.63±2.58 ^** IIl-Na 60.38±3.35 ^** Ie-Na 80.66±2.28 ^** IIm-Na 41.61±3.02 ^** If-Na 55.47±2.94 ^** IIn-Na 61.28±2.25 ^** Ig-Na 76.70±2.60 ^** IIo-Na 36.26±2.64 ^** Ih-Na 87.90±1.40 ^** IIp-Na 87.40±1.55 ^** Im-Na 81.66±1.44 ^** G-6-2 46.78±2.25 ^** In-Na 67.76±1.83 ^** GF-74 99.97±0.30 ^** Iq-Na 68.55±0.38 ^** GNP-h 0.00±1.38 Ir-Na 67.67±1.57 ^** G-g-c 19.93±2.13 ^** Is-Na 39.81±1.54 ^** GI-H3 99.24±2.00 ^** It-Na 62.20±1.88 ^** Glo-h 88.57±1.46 ^**

IIa 75.85±1.89 ^** GO-h 70.02±2.43 ^** GF-H3 83.50±5.69 ^** GLP-h 68.70±2.33 ^** GF-H2 100.00±2.44 ^** GFI-H3 79.14±3.45 ^** G-i 10.69±2.51 ^* GFN-H15 79.34±1.57 ^** GN-g1 77.31±2.30 ^** GF-H4 99.05±2.57 ^** GNO-7 83.36±1.44 ^** GLP-7 98.07±4.20 ^** G-8 86.41±4.47 ^** GF-3 70.00±2.05 ^** GF-73 67.36±3.87 ^** G-73 99.59±1.79 ^** GNO-h 72.53±2.77 ^** GN-72 75.22±2.89 ^** G-6-4 24.47±2.41 ^** GN-71 69.36±2.48 ^** GFO-715 69.09±2.41 ^** GNP-7 21.521.77 ^** GFO-74 70.56±2.27 ^** GO-7 73.63±4.45 ^** GF-72 100.00±2.48 ^** streptomycin sulfate 100.00±1.28 ^** blank group 0.00±0.00 Ampicillin 26.45±2.01 ^**

Compared with blank control group, ^* P<0.05; ^** P<0.01.

According to the above-mentioned activity screening results, the minimum inhibitory concentration (MIC) of the compound with better activity was further determined, and the determination results are shown in Tables 8, 9, and 10.

The MIC value of table 8 test compound to Bacillus subtilis

compound MIC (μM) G-6-4 50±5.5 ^** GF-72 30±5.5 ^** streptomycin sulfate 5±0.5

Compared with positive drug control group, **P<0.01.

Table 9 tested compounds to the MIC value of Staphylococcus aureus

compound MIC (μM) IIj 40±4.2 ^** G-6-4 10±2.0 G-i 4±2.0 GF-72 8±4.4 GF-H4 4±2.0 Il 10±1.1 GF-3 20±4.0 ^* Ampicillin 5±2.0

Compared with positive drug control group, ^* P<0.01; ^** P<0.01.

The MIC value of table 10 test compound to streptomyces scab

compound MIC (μM) IIm 50±2.0 ^** GF-H4 5±2.5 IIj 40±5.5 ^** GF-72 10±2.0 GF-H2 2±1.5 GLP-7 40±5.0 ^** G-73 5±2.5 GF-74 8±2.5 GI-H3 10±4.5 G-i 2±0.0 streptomycin sulfate 5±1.5

Compared with positive drug control group, ^** P<0.01.

Assay results show that compound G-6-4 and GF-72 show moderate inhibitory activity to Bacillus subtilis, but the activity is not as good as the positive control drug streptomycin sulfate (P<0.01); compound G-6-4, GF-72, G-i The inhibitory activity of GF-H4 and Il on Staphylococcus aureus was not significantly different from that of the positive control drug ampicillin, indicating that its antibacterial activity was equivalent to that of ampicillin, while the MIC value indicated that the inhibitory activity of G-i and GF-H4 was stronger than that of ampicillin penicillin. The inhibitory activity of compounds GF-H4, GF-H2, G-73, GF-74, G-i and GI-H3 on Streptomyces scabies was not significantly different from that of streptomycin sulfate, and the MIC value indicated that the compounds GF-H2 and G-i had The inhibitory activity is greater than that of streptomycin sulfate. The above results show that this type of glycyrrhetinic acid derivatives can significantly inhibit the growth of Gram-positive bacteria, and it is expected to be developed into antibacterial drugs and applied in the fields of agriculture and medicine.

5. Antifungal activity screening experiments of the glycyrrhetinic acid derivatives

Test compounds: Glycyrrhetinic acid (GA), Ia～Ix, Ia-Na, Ib-Na, Ie-Na, If-Na, Ig-Na, Ih-Na, Im-Na, In-Na, Iq- Na, Ir-Na, Is-Na, It-Na, IIa～IIp, IIa-Na～IIp-Na, G-73, GNP-h, GLP-7 and ester and amide derivatization in liver protection activity experiment substances, and the following structural compounds:

Experimental strains: Gibberella zeae, Fusarium oxyspoyum, and Phytophthora nicotiana were all provided by the Fungal Resources Research Laboratory of Guizhou University.

Main instruments and reagents: Fluorescence inverted phase-contrast microscope is a product of Nikon, Japan (model: Ti-U); automatic triple pure water distiller (SZ-97, Shanghai Yarong Biochemical Instrument Factory); ultra-clean bench (SW-CJ- 1F, Suzhou Purification Equipment Co., Ltd.); magnetic heating stirrer (C-MAG HS4, German IKA company); constant temperature shaking incubator (HZQ-F160, Jiangsu Taicang Experimental Equipment Factory); electronic balance (SL-114, American DENVER Company); micropipettes of various models (Germany eppendorf); 96-well culture plate is the product of Corning Costar Company in the United States; Model l550 microplate reader is the product of BIO-RAD Company in the United States; dimethyl sulfoxide, glucose, chlorothalonil , Carbendazim are products of sigma company.

Preparation of culture medium: Take 200g of peeled and washed potatoes, cut them into small pieces, put them into 1000mL of distilled water and boil for 30min, then filter with 4 layers of gauze to obtain the filtrate, add 20g of glucose to the filtrate, stir to dissolve, and make up for the missing water To 1000mL, aliquoted, stoppered and autoclaved to obtain a potato liquid medium (PDA medium), and 15%-20% agar was added to obtain a corresponding solid medium.

method:

Preparation of spore suspension: Under aseptic operation, activate the bacteria and inoculate them on the inclined PDA solid medium, cultivate them in a constant temperature incubator at 28°C, and place the inclined surface under strong light irradiation after the mycelia are covered with the medium. 2d to facilitate its sporulation. Microscopically observe its growth to maturity and produce abundant spores, add 5mL sterile saline to the test tube, gently scrape off the spores on the slope with an inoculation loop under aseptic conditions, and inhale the sterile saline containing spores Add sterile normal saline to the test tube and scrape the spores, repeat this operation several times, then combine the normal saline containing spores into the triangular flask, add 5 sterile glass beads with a diameter of 3mm to it , stopper the triangular flask, place it in a constant temperature oscillator and shake it at 200rpm for 1h, and finally filter it with sterile gauze to remove the mycelium to obtain the spore suspension.

The antifungal activity of the compounds was determined by the 96 microwell turbidity method. A test compound group (100 μmol/L), a positive drug control group (the concentrations of chlorothalonil and carbendazim are both 100 μmol/L) and a blank control group were set up. The compound is dissolved in PDA medium with DMSO to form a solution, and the DMSO concentration is not more than 1%. First, prepare the spore suspension with PDA liquid medium to make the spore concentration 2×10 ⁶ /mL, add the spore suspension to the 96-well plate, 135 μL per well, and then add 15 μL of the drug solution to each well, so that the final concentration of the drug is 100 μmol/mL L, the blank control group was replaced with the same volume of culture medium, each compound was set up with 3 duplicate wells, after the addition, the absorbance value A ₁ of each well was measured at 620nm with a microplate reader, and then the 96-well plate was placed at a constant temperature Culture in an incubator at 28°C. Observe the spore germination through a microscope. After the spores of the blank control group germinate, immediately measure the absorbance value A ₂ at 620 nm in each well again, calculate the difference ΔA between the two absorbance values before and after, and calculate the inhibition rate according to the following formula:

Inhibition rate = (blank group ΔA - drug group ΔA) * 100% / blank group ΔA

Each experiment was repeated three times, and the average inhibition rate was calculated.

Results: See Tables 11, 12 and 13 for the results of the inhibitory activity of the tested compounds against the three fungi.

The inhibitory effect of table 11 glycyrrhetinic acid derivatives on tobacco black shank bacteria (100 μ M)

compound Inhibition rate(%) compound Inhibition rate(%) GA 19.00±3.02 ^** IIm 7.51±4.28 Ia 16.55±0.48 ^** IIk 7.71±4.33 Ib 31.67±1.67 ^** IIj 28.82±3.50 ^** Ic 34.81±3.24 ^** IIo 18.33±2.77 ^** Id 15.66±2.67 ^** IIb 12.15±2.78 ^** I e 38.62±2.73 ^** IIc 46.38±2.12 ^** If 35.84±1.71 ^** IId 52.37±2.33 ^** Ig 63.22±1.28 ^** IIe 33.34±2.19 ^** Ih 46.53±3.15 ^** IIf 35.27±2.60 ^** Ii 21.53±1.62 ^** II g 6.81±2.38 Ij 46.63±2.88 ^** IIh 78.86±4.28 ^** Ik 46.76±2.68 ^** IIi 58.46±1.34 ^** Il 41.47±2.04 ^** IIl 36.47±1.58 ^** Im 52.70±2.10 ^** II n 65.42±2.38 ^** In 63.90±1.30 ^** IIp 64.28±1.70 ^** Io 71.63±1.94 ^** IIa-Na 70.45±3.25 ^** IP 63.76±1.33 ^** IIb-Na 56.36±5.07 ^** Iq 58.53±3.38 ^** IIc-Na 57.40±1.93 ^** Ir 78.67±1.97 ^** IId-Na 74.53±1.84 ^** Is 69.81±3.24 ^** IIe-Na 61.05±1.44 ^** it 54.36±4.44 ^** IIf-Na 36.28±4.37 ^** I u 6.65±2.07 IIg-Na 68.99±2.51 ^** IV 58.84±3.75 ^** IIh-Na 60.03±1.36 ^** Iw 58.62±1.58 ^** IIi-Na 77.48±1.37 ^** Ix 87.93±5.02 ^** IIj-Na 18.20±5.45 ^** Ia-Na 78.56±1.02 ^** IIk-Na 70.39±3.48 ^** Ib-Na 50.63±2.38 ^** IIl-Na 68.38±5.35 ^** Ie-Na 83.66±2.68 ^** IIm-Na 40.61±3.92 ^** If-Na 35.47±2.84 ^** IIn-Na 68.28±5.25 ^** Ig-Na 76.50±2.63 ^** IIo-Na 46.26±2.78 ^** Ih-Na 80.94±2.40 ^** IIp-Na 57.40±1.45 ^** Im-Na 71.66±4.44 ^** G-6-2 76.78±2.25 ^** In-Na 68.76±1.23 ^** GLO-7 76.98±1.08 ^** Iq-Na 58.55±0.38 ^** GNP-h 46.28±3.50 ^** Ir-Na 57.67±1.37 ^** G-g-c 23.12±3.45 ^** Is-Na 49.81±3.54 ^** GHI-73 19.49±2.81 ^** It-Na 65.20±1.68 ^** Glo-h 78.57±1.76 ^** IIa 29.68±1.90 ^** GO-h 73.02±2.13 ^** GF-H3 83.50±5.69 ^** GLP-h 48.70±2.33 ^** GF-H2 56.00±2.84 ^** GFI-H3 74.54±3.45 ^** G-i 70.69±2.31 ^** GFN-H15 11.19±2.44 ^* GN-g1 3.86±2.44 GF-H4 49.78±4.57 ^** GNO-7 77.96±3.40 ^** GLP-7 46.47±2.46 ^** G-8 76.41±4.37 ^** GF-H15 41.21±4.90 ^** GF-73 62.36±3.77 ^** G-73 52.56±3.27 ^** GNO-h 76.53±2.67 ^** GN-72 72.22±2.69 ^** G-6-4 16.48±2.88 ^** GN-71 59.36±2.68 ^**

GFO-715 62.09±2.21 ^** GNP-7 50.98±3.00 ^** GFO-74 71.56±2.77 ^** GO-7 70.63±4.05 ^** GF-72 70.00±2.38 ^** Carbendazim 80.84±3.01 ^** blank group 0.00±0.00 Chlorothalonil 98.83±2.81 ^**

Compared with blank control group, ^* P<0.01; ^** P<0.01.

Table 12 Inhibitory effect of glycyrrhetinic acid derivatives on Fusarium wilt of pepper (100 μM)

compound Inhibition rate(%) compound Inhibition rate(%) GA 17.57±4.00 ^** IIm 1.37±2.80 Ia 7.53±1.41 ^* IIk 8.47±2.80 ^* Ib 11.67±3.37 ^** IIj 10.98±2.38 ^* Ic 4.91±3.54 ^** II o 21.16±2.75 ^** Id 15.06±2.07 ^** IIb 29.23±3.28 ^** I e 28.62±2.70 ^** IIc 16.38±2.42 ^** If 25.11±1.01 ^** IId 12.37±4.33 ^** Ig 53.22±1.18 ^** IIe 33.54±4.19 ^** Ih 40.51±3.05 ^** IIf 45.27±4.61 ^** Ii 29.53±2.62 ^** IIg 32.16±2.70** Ij 45.63±2.98 ^** IIh 20.21±1.22 ^** Ik 36.76±2.08 ^** IIi 21.46±1.64 ^** Il 51.47±2.84 ^** IIl 39.47±2.58 ^** Im 12.71±4.80 ^** II n 45.47±2.34 ^** In 43.90±1.80 ^** IIp 24.28±4.78 ^** I o 61.63±2.95 ^** IIa-Na 37.45±3.75 ^** IP 23.76±1.73 ^** IIb-Na 26.36±5.37 ^** Iq 48.13±3.18 ^** IIc-Na 17.47±2.93 ^** Ir 58.17±1.37 ^** IId-Na 54.53±1.14 ^** Is 49.81±5.24 ^** IIe-Na 31.05±3.64 ^** it 50.16±4.84 IIf-Na 16.78±4.27 ^** I u 36.75±2.83 ^** IIg-Na 48.79±2.11 ^** IV 58.84±3.75 ^** IIh-Na 10.03±1.76 ^** Iw 8.62±1.38 IIi-Na 25.48±2.37 ^** Ix 27.93±4.02 ^** IIj-Na 38.20±3.45 ^** Ia-Na 28.16±3.02 ^** IIk-Na 30.59±3.46 ^** Ib-Na 30.33±2.33 ^** IIl-Na 10.45±2.35 ^* Ie-Na 23.69±2.60 ^** IIm-Na 20.61±3.72 ^** If-Na 34.48±2.14 ^** IIn-Na 38.78±5.21 ^** Ig-Na 36.51±4.63 ^** IIo-Na 19.18±2.70 ^** Ih-Na 30.13±3.58 ^** IIp-Na 17.49±2.42 ^** Im-Na 41.66±4.04 ^** G-6-2 36.18±2.28 ^** In-Na 48.46±5.23 ^** GLO-7 28.31±2.70 ^** Iq-Na 18.05±0.58 ^** GNP-h 36.27±4.07 ^** Ir-Na 2.50±1.70 G-g-c 19.57±1.78 ^** Is-Na 9.81±3.50 GHI-73 0.00±0.47 It-Na 29.64±3.55 ^** Glo-h 38.67±4.71 ^** IIa 9.95±3.31 ^* GO-h 3.02±2.93 GF-H3 43.50±5.49 ^** GLP-h 42.70±2.30 ^** GF-H2 6.09±2.14 GFI-H3 74.54±3.45 ^** G-i 30.69±2.39 ^** GFN-H15 16.15±2.45 ^**

GN-g1 23.06±2.54 ^** GF-H4 23.35±2.4 ^** GNO-7 6.60±2.77 GLP-7 6.41±2.56 G-8 56.48±4.33 ^** GF-H15 8.47±2.00 ^* GF-73 52.56±3.87 ^** G-73 7.30±2.55 GNO-h 6.53±2.17 ^** GN-72 2.25±2.79 G-6-4 9.29±4.18 ^* GN-71 19.36±2.62 ^** GFO-715 32.39±2.81 ^** GNP-7 10.19±3.79 ^* GFO-74 51.56±2.87 ^** GO-7 0.00±3.51 GF-72 10.00±2.18 ^* Carbendazim 98.54±3.22 ^** blank group 0.00±0.00 Chlorothalonil 100.0±1.05 ^**

Compared with blank control group, ^* P<0.01; ^** P<0.01.

Table 13 The inhibitory effect of glycyrrhetinic acid derivatives on wheat head blight (100μM)

compound Inhibition rate(%) compound Inhibition rate(%) GA 36.34±3.00 ^** IIm 2.82±1.08 Ia 8.23±4.87 ^* IIk 6.30±2.77 Ib 10.67±3.32 ^* IIj 11.85±2.57 ^* Ic 4.77±3.65 ^** II o 1.29±0.34 Id 5.66±3.87 ^** IIb 61.70±2.00 ^** I e 18.70±2.72 ^** IIc 1.38±2.46 ^** If 5.61±2.01 ^** IId 15.38±4.30 ^** Ig 13.92±2.10 ^** IIe 13.54±4.10 ^** Ih 10.58±3.75 ^* IIf 5.27±4.41 ^** Ii 21.58±2.60 ^** II g 3.79±2.47 Ij 15.60±2.58 ^** IIh 1.74±0.35 Ik 26.06±2.68 ^** IIi 2.46±1.65 Il 11.47±2.24 ^** IIl 9.47±2.08 ^* Im 2.71±4.68 ^** II n 0.08±2.84 In 4.95±1.85 ^** IIp 20.00±4.28** I o 21.63±2.55 ^** IIa-Na 7.45±3.45 ^* IP 3.76±2.73 IIb-Na 6.86±5.07 Iq 18.63±3.78 ^** IIc-Na 7.47±2.97 ^** Ir 11.33±4.44 ^* IId-Na 5.52±2.14 ^** Is 11.80±2.58 ^* IIe-Na 23.15±3.35 ^** it 20.16±4.44 ^** IIf-Na 2.10±1.59 I u 6.75±2.73 ^** IIg-Na 3.32±2.44 IV 5.37±3.85 ^** IIh-Na 11.21±2.95 ^* Iw 9.61±2.32 ^* IIi-Na 25.16±1.03 ^** Ix 11.65±4.02 ^** IIj-Na 0.00±2.00 Ia-Na 2.66±4.02 IIk-Na 25.87±1.33 ^** Ib-Na 10.83±3.30 ^* IIl-Na 1.75±4.35 Ie-Na 3.60±2.67 IIm-Na 0.79±2.90 If-Na 14.40±2.10 ^** IIn-Na 3.28±4.21 Ig-Na 16.51±4.03 ^** IIo-Na 9.25±2.80 ^* Ih-Na 0.00±3.28 IIp-Na 25.77±2.84 ^** Im-Na 4.00±4.38 G-6-2 36.18±2.28 ^** In-Na 8.40±5.13 ^* GLO-7 38.42±2.74 ^** Iq-Na 1.05±2.58 GNP-h 10.45±3.57 ^* Ir-Na 7.58±1.78 ^* G-g-c 5.13±1.46

Is-Na 0.81±3.57 GHI-73 21.76±3.70 ^** It-Na 0.00±0.64 Glo-h 18.64±4.51 ^** IIa 1.33±1.79 GO-h 3.92±2.56 GF-H3 13.55±5.40 ^** GLP-h 4.72±2.38 GF-H2 6.49±2.94 GFI-H3 7.37±3.00 ^* G-i 25.69±2.31 ^** GFN-H15 0.00±2.37 GN-g1 3.16±2.94 GF-H4 7.68±2.70 ^* GNO-7 19.69±0.08 ^** GLP-7 11.35±0.99 ^* G-8 16.48±4.03 ^** GF-H15 36.02±1.57 ^** GF-73 22.50±3.07 ^** G-73 1.47±1.05 GNO-h 26.55±2.77 ^** GN-72 8.25±2.69 ^* G-6-4 4.62±2.42 GN-71 2.22±2.08 GFO-715 0.77±3.69 GNP-7 5.22±2.70 GFO-74 11.58±2.27 ^* GO-7 0.61±3.33 GF-72 21.06±2.58 ^** Carbendazim 100.00±1.66 ^** blank group 0.00±0.00 Chlorothalonil 15.32±2.65 ^**

Compared with blank control group, ^* P<0.01; ^** P<0.01.

The test results showed that the tested compounds exhibited different degrees of inhibitory activity against the three fungi, among which the inhibitory effect on tobacco black shank was stronger. Compounds Ir, Ix, Ie-Na, Ih-Na, GF-H3, IIh, GNO -7 and GLO-7 inhibited significantly.

6. Anti-tobacco mosaic virus activity screening experiment of the glycyrrhetinic acid derivatives

Test compounds: Glycyrrhetinic acid (GA), Ia～Ix, Ia-Na, Ib-Na, Ie-Na, If-Na, Ig-Na, Ih-Na, Im-Na, In-Na, Iq- Na, Ir-Na, Is-Na, It-Na, IIa～IIp, IIa-Na～IIp-Na, GNP-h, GLP-7, GLO-7 and ester and amide derivatization in liver protection activity experiment things.

Tobacco mosaic virus (TMV) was provided by Guizhou Institute of Tobacco Science, and the test tobacco (Nicotinanglutinosa) was planted in the greenhouse of our laboratory, and the seedling age was 8 weeks.

Main instruments and reagents: positive control drug is 2% Ningnanmycin, Heilongjiang Qianger Biochemical Co., Ltd.; intelligent light incubator GXZ-280C, Zhejiang Ningbo Jiangnan Instrument Factory; various types of micropipettes (Germany eppendorf); Wen 20, Tianjin No. 3 Chemical Reagent Factory; Dimethyl sulfoxide (sigma); Sodium dihydrogen phosphate and disodium hydrogen phosphate were purchased from Tianjin Zhiyuan Chemical Reagent Co., Ltd.; Soft brushes, hard row pens, emery (320 mesh ) and brush medicine plate are commercially available.

The inactivation effect of the test compound on TMV in vitro, the preventive effect and the therapeutic effect on tobacco infection by TMV in vivo were respectively tested by half-leaf scorched spot method.

The inactivation effect of the test compound on TMV in vitro:

Experimental method: the test compound at a concentration of 500 μg/mL was co-incubated with the virus (6 μg/mL) for 30 min. Select the middle part of the heart leaf tobacco leaves with consistent growth as the treatment leaves (each plant selects 3-5 pieces with similar leaf positions, and each treatment repeats 3 plants), and utilizes the artificial rubbing method to inoculate the right half of all treatment leaves containing the test compound. TMV, while the left half lobe was inoculated with TMV without the test compound as a blank control. Then culture in a light incubator under the following conditions: daytime 08:00-22:00, temperature 25°C, light 10000 Lux; night 22:00-next day 08:00, temperature 22°C, no light. After culturing for 3-4 days, observe and record the number of scabs, and calculate the scab inhibition rate.

Blister inhibition rate = (C _{blank group} - C _{drug group} ) × 100% / C _{blank group}

In vivo preventive effect of test compounds against TMV infection in tobacco:

Experimental method: select the middle part of the open leaves of tobacco with consistent growth as the treatment leaves (each plant selects 3-5 sheets with similar leaf positions, and each treatment repeats 3 plants), and the right half leaf is sprayed with the compound solution for 24 hours before inoculating the virus. The left half lobe was replaced with 0.5% sodium carboxymethyl cellulose solution, and the test compounds were all dissolved in 0.5% sodium carboxymethyl cellulose solution with DMSO in advance, with a concentration of 500 μg/mL. 24 hours after applying the drug, inoculate the whole leaves of all treated leaves with TMV (6 μg/mL) by manual rubbing method, and then culture them in a light incubator. ; Dark night 22:00-08:00 the next day, the temperature is 22 ℃, no light. After culturing for 3-4 days, observe and record the number of scabs, and calculate the scab inhibition rate.

Therapeutic effects of test compounds on TMV-infected tobacco in vivo:

Experimental method: select the middle spread leaves of tobacco heart leaves with consistent growth as treatment leaves (3-5 pieces with similar leaf position are selected for each plant, and 3 plants are repeated for each treatment), and the whole leaves of all treatment leaves are inoculated with TMV by artificial rubbing method ( 6 μg/mL), 12 hours after virus inoculation, the right half leaf was sprayed with compound solution, and the left half leaf was replaced with 0.5% sodium carboxymethyl cellulose solution. The test compounds were all dissolved in 0.5% carboxymethyl cellulose in advance. In sodium cellulose solution, the concentration is 500μg/mL. After the drug is applied, the tobacco is placed in a light incubator for cultivation. The cultivation conditions are: 08:00-22:00 during the day, the temperature is 25°C, and the light is 10000Lux; 22:00-08:00 at night, the temperature is 22°C No light. After culturing for 3-4 days, observe and record the number of scabs, and calculate the scab inhibition rate.

Results: The test results are shown in Table 14, and see Figure 11. The results show that most of the tested compounds have preventive, therapeutic and in vitro passivation effects on TMV infection of tobacco. Among them, compounds Ig, Ig-Na, Ih-Na, Ib, Id, Iq, Iq-Na, Il, Ir-Na, Is-Na, IIb, IIc-Na, IIh-Na, IIk, IIj, G-6- 4. The therapeutic effects of GF-H2, GF-H3, GN-71, GO-h, GNP-h, GLO-7, and GNO-h on TMV-infected tobacco were significantly better than the positive control drug Ningnanmycin (P< 0.05), while compounds Ie-Na, In-Na, Ir-Na, Ir, IIl-Na, IIm, IIo, G-6-2, GF-H2, GO-h, GLO-7, GNO-h, GNO The preventive effect of -7 on tobacco infection by TMV was significantly better than that of Ningnanmycin (P<0.05). 73. The passivation effect of GO-h was significantly better than that of Ningnanmycin (P<0.05). Therefore, these compounds have significant anti-TMV activity and are expected to be developed as anti-TMV drugs and applied in agriculture.

Table 14 Inhibitory effect of glycyrrhetinic acid derivatives on tobacco mosaic virus (500 μg mL-1)

Compared with blank group, *P<0.05, **P<0.01; compared with positive control group, #P<0.05, ##P<0.01.

Compared with the prior art, the present invention provides a group of A-ring polyoxidized substituted glycyrrhetinic acid derivatives with novel structures and a preparation method thereof. The preparation method has mild reaction conditions, simple and easy operation, and does not require special reagents and reaction conditions , it is easy to expand the scale and realize the purpose of industrialized production; the polarity of some glycyrrhetinic acid derivatives prepared by the present invention is increased, so the solubility is significantly improved compared with glycyrrhetinic acid, which is conducive to improving the bioavailability of the compound and is more conducive to It is developed into medicines for the effective treatment of diseases; in addition, the compounds provided by the present invention and their pharmaceutically acceptable salts (sodium salts, potassium salts or calcium salts, etc.) have strong hepatoprotective activity and are effective against Gram Bacteria such as positive bacteria, fungi such as tobacco black shank bacterium and tobacco mosaic virus show strong inhibitory activity, and have good application prospects in the fields of medicine and pesticides.

Description of drawings

Fig. 1 is the observation diagram of the pathological changes of the liver tissue of the normal control group, the model group, and the silymarin group, wherein 1, 3, and 5 respectively represent the observation graphs of the pathological changes of the liver tissue of the normal control group, the model group, and the silymarin group enlarged by 200 times, 2, 4 and 6 respectively represent the magnified 400 times observation pictures of the pathological changes of the liver tissue in the normal control group, the model group and the silymarin group;

Fig. 2 is the observation diagram of the liver histopathological changes of compound G-8 low-dose group, middle-dose group and high-dose group;

Fig. 3 is the observation diagram of the pathological changes of the liver tissue in the compound Ⅱb low-dose group, middle-dose group and high-dose group;

Fig. 4 is the observation diagram of the liver histopathological changes of compound GNP-7 low dose group, middle dose group and high dose group;

Fig. 5 is the observation diagram of the pathological changes of the liver tissue in the compound Ic low-dose group, middle-dose group and high-dose group;

Fig. 6 is the observation diagram of the pathological changes of the liver tissue in the compound IIj low-dose group, middle-dose group and high-dose group;

Fig. 7 is the observation diagram of the liver histopathological changes of compound G-i low-dose group, middle-dose group and high-dose group;

Fig. 8 is the observation diagram of the pathological changes of the liver tissue in the compound IId low-dose group, middle-dose group and high-dose group;

Figure 9 is an observation diagram of the pathological changes of liver tissue in the compound G-6-2 low-dose group, middle-dose group and high-dose group;

Figure 10 is an observation diagram of the pathological changes of liver tissue in the compound GN-g1 low-dose group, middle-dose group and high-dose group;

In Figures 2 to 10, 1, 3, and 5 represent the 200-fold magnified observation images of the pathological changes of the liver tissue in the low, medium, and high dose groups, respectively, and 2, 4, and 6 represent the liver tissue in the low, medium, and high dose groups, respectively. The pathological changes are observed at 400 times magnification.

Fig. 11 is the preventive effect of GLO-7, Ⅱj on tobacco infection TMV, therapeutic effect, in vitro to the inactivation effect figure of TMV; Wherein A is the therapeutic effect of Ningnanmycin on tobacco infected with TMV, B is the effect of GLO-7 on tobacco infection Preventive effect of tobacco infected with TMV, C is the inactivation effect of Ⅱj on TMV in vitro, D is the therapeutic effect of Ⅱj on tobacco infected with TMV.

Detailed ways

Embodiment 1 of the present invention: preparation of compound G-1:

Take 3.00g (6.38mmol) glycyrrhetinic acid in a 250ml round bottom flask, add 100ml N,N-dimethylformamide (DMF) to dissolve it completely, add 1.00g (7.25mmol) potassium carbonate powder, After stirring for 15 minutes, 7.0 mmol of benzyl bromide was added, and the reaction was refluxed under nitrogen protection. TLC (thin layer chromatography) was used to detect the progress of the reaction (the developer was V (petroleum ether): V (acetone) = 5:1) until the reaction was completed. After 2 hours of reaction, the reaction liquid was extracted with ethyl acetate after adding water. The organic layer was first adjusted to pH 7 with saturated _NaHCO3 solution, then washed with saturated NaCl solution and dried with anhydrous magnesium sulfate, then filtered, and the solvent was evaporated under reduced pressure to obtain a yellow color. Crude oil. Silica gel column chromatography (V (petroleum ether): V (ethyl acetate) = 8: 1) yielded 3.52 g of compound G-1 with a yield of 98.5%.

Compound G-1, white powder, C ₃₇ H ₅₂ O ₄ , m.p133～136℃; IR(KBr),σ/cm ^-1 :3423,2930,2870,1725,1654,1466,1386,1213,1149 , 1083, 1037, 824, 733, 697; ¹ H NMR (400MHz, CDCl ₃ ) δ: 7.38-7.33 (m, 5H, H-Ar), 5.54 (s, 1H, H-12), 5.20 (d, 1H, J= 12.0Hz, Bn-CHH'), 5.09(d, 1H, J＝12.0Hz, Bn-CHH'), 3.22(dd, 1H, J＝5.6Hz, 10.4Hz, H-3), 2.78(dt, 1H ,J＝13.2Hz,H-1),2.31(s,1H,H-9),1.34(s,3H,H-27),1.16(s,3H,H-29),1.13(s,3H, H-25),1.10(s,3H,H-23),1.00(s,3H,H-26),0.80(s,3H,H-24),0.73(s,3H,H-28); ¹³ C NMR (100MHz, CDCl ₃ )δ: 200.1(C-11), 176.2(C-30), 169.0(C-13), 136.0(C _ar ), 128.5(C _ar ), 128.5(C _ar ), 128.4 (C-12),128.2(C _ar ),127.2(C _ar ),127.2(C _ar ),78.7(C-3),66.1(Bn-CH ₂ ),61.7(C-9),54.8(C- 5), 48.1(C-18), 45.3(C-14), 43.9(C-20), 43.1(C-8), 41.0(C-19), 39.0(C-1), 39.0(C-4 ),37.6(C-22),37.0(C-10),32.7(C-7),31.7(C-17),31.1(C-21),28.3(C-29),28.2(C-28) ,28.0(C-23),27.2(C-2),26.4(C-16),26.3(C-15),23.3(C-27),18.6(C-26),17.4(C-6), 16.3 (C-25), 15.5 (C-24); EI (m/z): 560 (M), 91 (100), 545, 469, 393, 352, 225.

Embodiment 2: the preparation of compound G-b:

Take 3.00g (5.35mmol) of compound G-1 in a 500ml round bottom flask, dissolve it in 200ml of absolute ethanol, add 40.00g of zinc powder, stir in an ice bath, and slowly drop in 100ml of hydrochloric acid until the reaction is complete. After 5 hours of reaction, the unreacted zinc powder was removed by filtration, and the filtrate was poured into 500ml of ice water and left to stand for 2 hours. The precipitate was precipitated, and the white precipitate was obtained by suction filtration, and recrystallized from methanol to obtain 2.40 g of compound G-b, with a yield of 82.2%.

Compound Gb, colorless needles, C ₃₇ H ₅₄ O ₃ , mp160～162℃; IR(KBr),σ/cm ^-1 :3424,2926,2871,1727,1638,1454,1383,1313,1215,1151 , 1085, 1044, 1029, 995, 740, 695; ¹ H NMR (400MHz, CDCl ₃ ) δ: 7.36-7.30 (m, 5H, H-Ar), 5.17 (brt, 1H, J=3.6Hz, H-12), 5.18 (d, 1H, J=12.0Hz, Bn-CHH'), 5.08(d, 1H, J=12.0Hz, Bn-CHH'), 3.22(dd, 1H, J=5.2Hz, 10.8Hz, H-3 ),1.18(s,3H,H-27),1.14(s,3H,H-29),0.99(s,3H,H-25),0.94(s,3H,H-23),0.93(s, 3H, H-26), 0.79(s, 3H, H-24), 0.73(s, 3H, H-28); ¹³ C NMR (100MHz, CDCl ₃ ) δ: 176.9(C-30), 144.2(C -13),136.3(C _ar ),128.4(C _ar ),128.4(C _ar ),128.0(C _ar ),127.9(C _ar ),127.9(C _ar ),122.5(C-12),78.9(C ar -3), 65.9(B n-CH ₂ ), 55.1(C-5), 48.0(C-18), 47.5(C-9), 44.2(C-20), 42.7(C-19), 41.4( C-14), 39.7(C-8), 38.7(C-1), 38.5(C-22), 38.2(C-4), 36.8(C-10), 32.6(C-7), 31.8(C -17), 31.2(C-21), 28.5(C-29), 28.1(C-28), 28.0(C-23), 27.1(C-2), 26.8(C-16), 26.0(C- 15), 25.9(C-27), 23.45(C-11), 18.3(C-6), 16.7(C-26), 15.5(C-25), 15.5(C-24); EI(m/z ):544(M ⁺ -2H),247(100),437,375,338,91,69,43.

Embodiment 3: the preparation of compound G-2:

Take 2.20g (3.93mmol) of compound G-1 in a 250ml round bottom flask, add 100ml of acetone to dissolve, stir in an ice bath for 15min, then slowly add 10.68ml (4.00mmol) of the Jones reagent that is now prepared, react at room temperature under nitrogen protection, TLC Detection of reaction progress (V (petroleum ether): V (ethyl acetate) = 6: 1), 2h reaction is complete. Use 2mol/L sodium hydroxide solution to adjust the pH of the reaction solution to neutral to terminate the reaction, evaporate acetone under reduced pressure, add water to disperse, and extract with ethyl acetate. The organic layer is washed with saturated _NaHCO3 solution and saturated NaCl solution successively Dry over magnesium sulfate and filter, and distill off the solvent under reduced pressure to obtain the crude product as a colorless powder. Recrystallization from methanol gave 22.11 g of compound G-2 with a yield of 96.4%.

Compound G-2, colorless flaky crystal, C ₃₇ H ₅₀ O ₄ , m.p89～91℃; IR(KBr),σ/cm ^-1 :2952,2869,1726,1705,1654,1455,1385, 1211,1148,1110,1085,742,730,698; ¹ H NMR (400MHz, CDCl ₃ )δ:7.38-7.34(m,5H,H-Ar),5.58(s,1H,H-12),5.20(d,1H ,J=12.4Hz,Bn-CHH'),5.09(d,1H,J=12.4Hz,Bn-CHH'),2.96(m,1H,H-2a),2.63(m,1H,H-2b) ,2.42(s,1H,H-9),2.36(brd,1H,J=16Hz,H-18),1.35(s,3H),1.26(s,3H),1.16(s,3H),1.15( s,3H),1.10(s,3H),1.06(s,3H),0.74(s,3H); ¹³ C-NMR(100MHz,CDCl ₃ )δ:217.2(C-3),199.3(C-11 ),176.1(C-30),169.53(C-13),136.06(C _ar ),128.5(C _ar ),128.5(C _ar ),128.3(C-12),128.2(C _ar ),128.2(C ar _ar ), 128.2(C _ar ), 66.2(Bn-CH ₂ ), 60.9(C-9), 55.3(C-5), 48.1(C-18), 47.7(C-4), 45.1(C-14 ), 43.9(C-20), 43.2(C-8), 41.0(C-19), 39.7(C-1), 37.5(C-22), 36.6(C-10), 34.1(C-7) ,32.0(C-21),31.7(C-17),31.1(C-2),28.4(C-29),28.2(C-28),26.4(C-15),26.3(C-16), 26.3(C-23), 23.2(C-27), 21.3(C-24), 18.7(C-6), 18.4(C-26), 15.6(C-25); EI(m/z):558 (M ⁺ ), 91(100), 543, 467, 393, 352, 225.

Embodiment 4: the preparation of compound G-c:

Compound G-c was prepared using compound G-b as a raw material, and the method was the same as that of compound G-2, with a yield of 96.8%.

Compound Gc, white needle crystal, C ₃₇ H ₅₂ O ₃ , m.p80～83℃; IR(KBr),σ/cm ^-1 :2958,2922,1730,1707,1455,1384,1213,1149,1084, 1044,744,696; ¹ H NMR (400MHz, CDCl ₃ )δ: 7.37-7.30 (m, 5H, H-Ar), 5.18 (t, 1H, J=3.2Hz, H-12), 5.19 (d, 1H, J＝12.4Hz, Bn-CHH'), 5.08(d, 1H, J＝12.4Hz, Bn-CHH'), 2.55(m, 1H, H-2a), 2.47(m, 1H, H-2b), 1.15(s,3H,H-29),1.13(s,3H,H-27),1.09(s,3H,H-23),1.07(s,3H,H-25),1.05(s,3H, H-24),1.00(s,3H,H-26),0.74(s,3H,H-28); ¹³ C NMR(100MHz,CDCl ₃ )δ:176.9(C-30),144.2(C-13 ),136.3(C _ar ),128.4(C _ar ),128.4(C _ar ),128.0(C _ar ),127.9(C _ar ),127.9(C _ar ),122.5(C-12),78.9(C-3 ), 65.9(Bn-CH ₂ ), 55.1(C-5), 48.0(C-18), 47.5(C-9), 44.2(C-4), 42.7(C-19), 41.4(C-20 ),39.7(C-14),38.7(C-1),38.5(C-22),38.2(C-8),36.8(C-10),32.6(C-2),31.8(C-17) ,31.2(C-7),28.5(C-29),28.1(C-28),28.0(C-23),27.1(C-21),26.8(C-16),26.0(C-15), 25.9(C-27), 23.4(C-11), 18.3(C-6), 16.7(C-24), 15.5(C-26), 15.5(C-25); EI(m/z):544 (M ⁺ ), 247(100), 437, 375, 338, 91.

Embodiment 5: the preparation of compound G-3:

Take 2.00g (3.58mmol) of compound G-2 in a 250ml round bottom flask, add 100ml of acetic anhydride to dissolve it completely, then add 0.43g (4.30mmol) of SeO ₂ , reflux reaction under nitrogen protection, TLC detection reaction process (expanded The agent is V (petroleum ether): V (acetone) = 10: 1) until the reaction is completed. 2.5h after the reaction is complete, add 2mol/L sodium hydroxide solution to the reaction solution to adjust the pH of the reaction solution to neutral, then extract with ethyl acetate, wash the organic layer with saturated NaCl solution and dry over anhydrous magnesium sulfate, filter, and evaporate under reduced pressure Solvent was used to obtain a crude brown oil. Silica gel column chromatography (V (petroleum ether): V (ethyl acetate) = 10:1) yielded 0.90 g of compound G-3 with a yield of 45.3%.

Compound G-3, yellow powder, C ₃₇ H ₄₈ O ₄ , m.p187～190℃; IR(KBr),σ/cm ^-1 :3062,3033,2968,2867,1727,1652,1615,1462,1386 , 1212, 1162, 766, 750, 716; ¹ H NMR (400MHz, CDCl ₃ ) δ: 7.73 (d, 1H, J=10Hz, H-1), 7.38-7.34 (m, 5H, H-Ar), 5.79 (d, 1H, J=10Hz, H-2), 5.64(s, 1H, H-12), 5.22(d, 1H, J=12.4Hz, Bn-CHH'), 5.09(d, 1H, J=12.4Hz, Bn-CHH'),2.65(s,1H,H-9),1.41(s,3H),1.37(s,3H),1.17(s,3H),1.16(s,3H),1.16(s,3H ), 1.11(s,3H), 0.75(s,3H); ¹³ C NMR (100MHz, CDCl ₃ )δ: 204.6(C-3), 198.9(C-11), 176.1(C-30), 170.5( C-13),161.6(C-1),136.0(C _ar ),128.5(C _ar ),128.5(C _ar ),128.3(C _ar ),,128.2(C _ar ),128.2(C _ar ),128.0 (C-2), 124.5(C-12), 66.2(Bn-CH ₂ ), 52.5(C-9), 52.7(C-5), 48.2(C-18), 45.4(C-4), 44.7 (C-14), 43.9(C-20), 43.4(C-10), 41.0(C-19), 38.7(C-8), 37.5(C-22), 31.9(C-7), 31.7( C-17),31.0(C-21),28..4(C-29),28.2(C-28),27.5(C-23),26.4(C-15),26.2(C-16), 23.3 (C-27), 21.5 (C-24), 20.0 (C-25), 18.8 (C-26), 18.1 (C-6); EI (m/z): 556 (M ⁺ ), 91 ( 100), 541, 487, 468, 419.

Embodiment 6: the preparation of compound G-d:

Compound G-d was prepared from compound G-c by the same method as that of compound G-3, with a yield of 50.3%.

Compound Gd, yellow powder, C ₃₇ H ₅₀ O ₃ , m.p66～69℃; IR(KBr), σ/cm ^-1 :3442,2929,1728,1668,1455,1382,1214,1155,1083,824,733,697 ; ¹ H NMR (400MHz, CDCl ₃ )δ:7.38-7.34(m,5H,H-Ar),7.04(d,1H,J=10.4Hz,H-1),5.81(d,1H,J=10.4 Hz,H-2),5.22(brt,1H,H-12),5.20(d,1H,J=12.4Hz,Bn-CHH'),5.09(d,1H,J=12.4Hz,Bn-CHH' ),1.18(s,3H,H-29),1.15(s,3H,H-25),1.14(s,3H,H-23),1.13(s,3H,H-27),1.10(s, 3H, H-24), 1.04(s, 3H, H-26), 0.75(s, 3H, H-28); ¹³ C NMR (100MHz, CDCl ₃ ) δ: 205.3(C-3), 176.8(C -30),159.2(C-1),144.8(C-13),136.3((C _ar ),128.4(C _ar ),128.4(C _ar ),128.0(C _ar ),128.0(C _ar ),128.0 (C _ar ),124.9(C-2),121.7(C-12),65.9(Bn-CH ₂ ),53.3(C-5),48.2(C-18),44.5(C-4),44.2( C-20), 42.5(C-19), 41.8(C-9), 41.6(C-10), 40.5(C-8), 39.3(C-14), 38.1(C-22), 32.3(C -17), 31.9(C-7), 31.2(C-21), 28.5(C-29), 28.1(C-28), 27.8(C-23), 26.8(C-16), 26.0(C- 15), 25.8(C-27), 23.4(C-11), 21.6(C-24), 18.9(C-25), 18.8(C-6), 17.3(C-26); EI(m/z ):542(M ⁺ ),91(100),527,451,405,338,247.

Embodiment 7: the preparation of compound Ic, Io:

Dissolve 800.00 mg (1.44 mmol) of compound G-3 in 250 ml of methanol, add 690 μL (1.73 mmol) of 10% NaOH solution, stir at room temperature for 30 min, then add 195 μL (1.73 mmol) of 30% H2O2 solution, react at room temperature under nitrogen protection, and detect by TLC The reaction process (developing agent is V (petroleum ether): V (acetone) = 10: 1) until the reaction is completed. 8h after the reaction is complete, add 2mol/L hydrochloric acid solution to the reaction solution to adjust the pH of the reaction solution to neutral to terminate the reaction, distill methanol off under reduced pressure, add water and extract with ethyl acetate, and the organic layer is sequentially washed with saturated NaHCO ₃ solution and saturated NaCl After the solution was washed and dried over anhydrous magnesium sulfate, it was filtered, and the solvent was distilled off under reduced pressure to obtain a crude white powder. Silica gel column chromatography (V (petroleum ether): V (acetone) = 15: 1) yielded 726.73 mg of compound Ic and 1.12 mg of Io301.12 mg, with yields of 88.3% and 42.8%, respectively.

Compound Ic, white powder, C ₃₇ H ₄₈ O ₅ , _m.p 140～143°C; IR(KBr), σ/cm ^-1 :2955, 2866, 1731, 1698, 1649, 1466, 1389, 1216, 1160, 1085,865,750,696; ¹ H NMR (400MHz, CDCl ₃ )δ:7.41-7.33(m,5H,H-Ar),5.64(s,1H,H-12),5.21(d,1H,J＝12.4Hz, Bn-CHH'),5.10(d,1H,J=12.4Hz,Bn-CHH'),4.49(d,1H,J=4.4Hz,H-2),3.38(d,1H,J=4.4Hz, H-1),2.90(s,1H,H-9),1.41(s,3H),1.21(s,3H),1.17(s,3H),1.13(s,3H),1.11(s,3H) ,1.02(s,3H),0.75(s,3H); ¹³ C NMR (100MHz,CDCl ₃ )δ:212.4(C-3),198.9(C-11),176.1(C-30),170.7(C -13),136.0(C _ar ),128.5(C _ar ),128.5(C _ar ),128.3(C _ar ),128.2(C _ar ),128.2(C _ar ),127.9(C-12),66.2(Bn -CH ₂ ), 64.6(C-1), 57.2(C-2), 54.5(C-9), 48.2(C-18), 45.5(C-5), 45.1(C-4), 44.8(C -14), 43.9(C-20), 43.4(C-10), 41.0(C-19), 38.3(C-8), 37.5(C-22), 31.7(C-7), 31.6(C-17 ), 31.0(C-21), 28.4(C-29), 28.2(C-28), 27.9(C-23), 26.5(C-15), 26.2(C-16), 23.2(C-27) , 20.9 (C-24), 18.6 (C-25), 18.1 (C-6), 15.8 (C-26); EI (m/z): 572 (M ⁺ ), 91 (100), 557, 481, 463, 449, 435.

Compound Io, white powder, C ₃₇ H ₄₈ O ₅ , m.p 124～126°C; ¹ H NMR (400MHz, CDCl ₃ ) δ: 7.41-7.33(m,5H,H-Ar), 5.62(s,1H, H-12),5.20(d,1H,J=12.4Hz,Bn-CHH'),5.08(d,1H,J=12.4Hz,Bn-CHH'),4.39(d,1H,J=4.0Hz, H-2),3.38(d,1H,J＝3.9Hz,H-1),2.90(s,1H,H-9),1.45(s,3H),1.23(s,3H),1.21(s, 3H),1.16(s,3H),1.12(s,3H),1.02(s,3H),0.90(s,3H); EI(m/z):572(M ⁺ ),91(100),557,481,463,449 .

Embodiment 8: the preparation of compound Id, Ip:

Compounds Id and Ip were prepared using compounds G-d as raw materials, and the method was the same as that of compounds Ic and Io.

Compound Id, yield 90.1%, white powder, C ₃₇ H ₅₀ O ₄ , m.p88～90℃; IR(KBr), σ/cm ^-1 :2956,1727,1699,1455,1383,1212,1155, 1084,732,697; ¹ H NMR (400MHz, CDCl ₃ )δ:7.37-7.26(m,5H,H-Ar),5.22(brs,1H,H-12),5.21(d,1H,J＝12.4Hz, Bn-CHH'),5.08(d,1H,J=12.4Hz,Bn-CHH'),3.51(d,1H,J=4.4Hz,H-1),3.38(d,1H,J=4.4Hz, ¹³ C NMR (100MHz, CDCl ₃ )δ:212.9(C-3),198.9(C-11),176.9(C-30),144.8(C-13),136.3(C _ar ),128.4(C _ar ),128.4( C _ar ),128.0(C _ar ),128.0(C _ar ),128.0(C _ar ),121.7(C-12),65.9(Bn-CH ₂ ),63.9(C-1),56.9(C-2) ,48.2(C-18),45.9(C-5),44.8(C-4),44.2(C-20),42.6(C-19),41.7(C-8),40.5(C-9), 39.9(C-14), 38.4(C-10), 38.2(C-22), 32.0(C-17), 31.9(C-7), 31.2(C-21), 28.5(C-29), 28.1 (C-28),27.9(C-23),26.8(C-16),26.1(C-15),25.7(C-27),23.9(C-11),20.9(C-24),18.8( C-6), 17.1 (C-26), 15.1 (C-25); EI (m/z): 558 (M ⁺ ), 91 (100), 543, 467, 421, 338, 247.

Compound Ip, yield 93.5%, white powder, C ₃₇ H ₅₀ O ₄ , m.p 109～110°C; ¹ H NMR (400MHz, CDCl ₃ ) δ: 7.35-7.25 (m, 5H, H-Ar), 5.22 (brs,1H,H-12),5.21(d,1H,J=12.4Hz,Bn-CHH'),5.08(d,1H,J=12.4Hz,Bn-CHH'),3.11(d,1H, J＝4.0Hz, H-1), 3.05(d, 1H, J＝4.0Hz, H-2), 1.18(s, 3H), 1.16(s, 3H), 1.09(s, 3H), 1.07(s ,6H), 0.87(s,3H), 0.72(s,3H); EI(m/z):558(M ⁺ ),91(100),467,338,247,45.

Embodiment 9: the preparation of compound Ii, Ij, Iu, Iv:

Take 500.00 mg (874.12 mmol) of compound Ic and dissolve it in 200 ml of methanol, stir in an ice bath for 15 min, then slowly add 332.12 mg (8.74 mol) of NaBH ₄ powder, react in an ice bath under the protection of nitrogen, and detect the reaction progress by TLC (developing agent is V (petroleum ether): V (acetone) = 5:1) until the reaction is complete. 1.5h after the reaction is complete, add 2mol/L hydrochloric acid solution to the reaction solution to adjust the pH to neutral to terminate the reaction, distill methanol off under reduced pressure, add water and extract with ethyl acetate, and use saturated NaHCO ₃ solution and saturated NaCl solution in sequence for the organic layer After washing and drying with anhydrous magnesium sulfate, filter, and distill off the solvent under reduced pressure to obtain a crude white powder. Compounds Ii, Ij, Iu and Iv were obtained by silica gel column chromatography (V (petroleum ether): V (acetone) = 6:1), with yields of 77.4%, 50.6%, 39.6%, and 27.1%, respectively.

Compound Ii, white powder, C ₃₇ H ₅₀ O ₅ , m.p81～84℃; IR(KBr),σ/cm ^-1 :3446,2957,2868,1728,1658,1455,1386,1215,1160,826,751,697 ; ¹ H NMR (400MHz, CDCl ₃ )δ:7.40-7.33(m,5H,H-Ar),5.59(s,1H,H-12),5.21(d,1H,J＝12.4Hz,Bn-CHH '),5.09(d,1H,J=12.4Hz,Bn-CHH'),4.10(d,1H,J=2.8Hz,H-3),3.55(d,1H,J=5.6Hz,H-2 ),3.05(d,1H,J＝3.6Hz,H-1),2.79(s,1H,H-9),1.36(s,3H),1.30(s,3H),1.16(s,3H), 1.12(s,3H),0.94(s,3H),0.81(s,3H),0.73(s,3H); ¹³ C NMR(100MHz,CDCl ₃ )δ:199.9(C-11),176.2(C- 30),169.8(C-13),136.0(C _ar ),128.6(C _ar ),128.6(C _ar ),128.2(C _ar ),128.1(C _ar ),128.1(C _ar ),128.0(C- 12), 75.3(C-3), 66.2(Bn-CH ₂ ), 61.2(C-1), 57.9(C-2), 56.1(C-5), 48.2(C-18), 45.2(C- 14), 44.5(C-9), 43.9(C-20), 43.3(C-8), 40.9(C-19), 37.6(C-22), 36.9(C-10), 36.3(C-4 ), 32.4(C-7), 31.7(C-17), 31.1(C-21), 28.3(C-29), 28.2(C-28,C-23), 26.4(C-15), 26.3( C-16), 23.2(C-27), 18.9(C-24), 17.9(C-26), 17.8(C-25), 16.0(C-6); EI(m/z): 574(M ⁺ ),91(100),559,491,352,313,135.

Compound Ij, white powder, C ₃₇ H ₅₀ O ₅ , m.p163～165℃; IR(KBr),σ/cm ^-1 :3538,2984,2922,2873,1720,1649,1455,1310,1214,1155 ,1041,984,871,740,697; ¹ H NMR (400MHz, CDCl ₃ )δ:7.40-7.33(m,5H,H-Ar),5.59(s,1H,H-12),5.21(d,1H,J＝12.4Hz ,Bn-CHH'),5.09(d,1H,J=12.0Hz,Bn-CHH'),4.29(d,1H,J=4.0Hz,H-3),3.54(brs,1H,H-2) ,3.42(t,1H,J=4Hz,H-1),2.80(s,1H,H-9),1.39(s,3H),1.28(s,3H),1.17(s,3H),1.11( s,3H),0.89(s,3H),0.82(s,3H),0.74(s,3H); ¹³ C NMR(100MHz,CDCl ₃ )δ:199.8(C-11),176.2(C-30) ,169.9(C-13),136.1(C _ar ),128.6(C _ar ),128.6(C _ar ),128.2(C _ar ),128.1(C _ar ),128.1(C _ar ),128.0(C-12) ,72.5(C-3),66.2(Bn-CH ₂ ),63.1(C-1),56.2(C-2),55.9(C-5),48.3(C-18),45.3(C-14) ,43.9(C-20),43.3(C-8),41.7(C-9),40.9(C-19),37.6(C-22),37.3(C-10),36.4(C-4), 32.4(C-7), 31.7(C-17), 31.1(C-21), 28.4(C-29), 28.2(C-28), 26.4(C-15), 26.3(C-16), 25.8 (C-23), 23.7(C-27), 23.3(C-24), 18.9(C-26), 17.8(C-25), 16.6(C-6); EI(m/z): 575( M ⁺⁺ H), 91(100), 560, 491, 393, 352, 225, 135.

Compound Iu, white powder, C ₃₇ H ₅₀ O ₅ , m.p114～116℃; ¹ H NMR (400MHz, CDCl ₃ )δ: 7.41-7.33(m,5H,H-Ar), 5.57(s,1H, H-12),5.20(d,1H,J=12.4Hz,Bn-CHH'),5.09(d,1H,J=12.0Hz,Bn-CHH'),4.19(d,1H,J=4.0Hz, H-3),3.50(brs,1H,H-2),3.40(t,1H,J=4.0Hz,H-1),2.82(s,1H,H-9),1.32(s,3H), 1.25(s,3H),1.17(s,3H),1.08(s,3H),0.88(s,3H),0.82(s,3H),0.64(s,3H);EI(m/z):576 (M ⁺ ),91(100),560,491,393,225,135,43.

Compound Ⅰv, white powder, C ₃₇ H ₅₀ O ₅ , m.p100～102℃; ¹ H NMR (400MHz, CDCl ₃ )δ: 7.41-7.35(m,5H,H-Ar), 5.55(s,1H, H-12),5.20(d,1H,J=12.4Hz,Bn-CHH'),5.10(d,1H,J=12.0Hz,Bn-CHH'),4.14(d,1H,J=3.6Hz, H-3),3.56(brs,1H,H-2),3.42(t,1H,J=4.0Hz,H-1),2.80(s,1H,H-9),1.33(s,3H), 1.21(s,3H),1.14(s,3H),1.10(s,3H),0.98(s,3H),0.88(s,3H),0.69(s,3H); EI(m/z):576 (M ⁺ ),91(100),560,491,393,225,135,43.

Embodiment 10: the preparation of compound Ik, Il, Iw, Ix:

Compounds Ik, Il, Iw and Ix were prepared from compound Id by the same method as that of compounds Ii, Ij, Iu and Iv. The yields were 79.4%, 44.8%, 31.2% and 18.6%, respectively.

Compound Ik, white powder, C ₃₇ H ₅₂ O ₄ , m.p90～92℃; IR(KBr),σ/cm ^-1 :3442,2955,1729,1658,1455,1381,1214,1148,1082,1049 , 1027,752,697; ¹ H NMR (400MHz, CDCl ₃ ) δ: 7.37-7.32 (m, 5H, H-Ar), 5.20 (d, 1H, Bn-CHH', J=12.0Hz), 5.18 (brs, 1H,H-12),5.08(d,1H,Bn-CHH',J=12.8Hz),3.57(s,1H,H-3),3.07(d,J=4.0Hz,1H,H-2) ,2.99(d,1H,J＝3.6Hz,H-1),1.15(s,3H),1.14(s,3H),1.13(s,3H),0.97(s,3H),0.95(s,3H ), 0.79(s,3H), 0.74(s,3H); ¹³ C NMR (100MHz, CDCl ₃ )δ: 176.9(C-30), 144.4(C-13), 136.4(C _ar ), 128.4(C _ar ),128.4(C _ar ),128.0(C _ar ),127.9(C _ar ),127.9(C _ar ),121.9(C-12),75.8(C-3),65.9(Bn-CH ₂ ),60.3 C-1), 57.3(C-2), 48.1(C-18), 45.5(C-5), 44.2(C-20), 42.7(C-19), 41.5(C-10), 41.1(C -9), 39.7(C-8), 38.2(C-22), 36.9(C-14), 36.4(C-4), 32.3(C-7), 31.8(C-17), 31.3(C- 15), 28.5(C-23), 28.4(C-27), 28.1(C-28), 26.8(C-16), 26.1(C-21), 25.8(C-29), 23.6(C-11 ),18.0(C-24),16.9(C-25),16.8(C-6),16.6(C-26); EI(m/z):558(M ⁺ -2H),247(100), 542, 451, 423, 405, 338, 247, 91, 69, 43.

Compound Il, white powder, C ₃₇ H ₅₂ O ₄ , m.p96～98℃; IR(KBr),σ/cm ^-1 :3432,2956,1730,1455,1382,1212,1152,1085,1046,1027 ,999,736,696; ¹ H NMR (400MHz, CDCl ₃ )δ: 7.35-7.29 (m, 5H, H-Ar), 5.18 (d, 1H, J=12.4Hz, Bn-CHH'), 5.16 (brs, 1H, H-12), 5.06(d, 1H, J=12.4Hz, Bn-CHH'), 3.51(brs, 1H, H-3), 3.42(t, 1H, J=4.4Hz, H-2), 3.16 (d,1H,J=4.0Hz,H-1),1.14(s,3H),1.13(s,3H),1.07(s,3H),0.94(s,3H),0.87(s,3H), 0.78(s,3H),0.72(s,3H); ¹³ C NMR(100MHz,CDCl ₃ )δ:176.9(C-30),144.6(C-13),136.4(C _ar ),128.4(C _ar ) ,128.4(C _ar ),127.9(C _ar ),127.9(C _ar ),127.9(C _ar ),121.8(C-12),72.7(C-3),65.9(Bn-CH ₂ ),62.3(C -1), 55.4(C-2), 48.1(C-18), 44.2(C-20), 42.7(C-19), 42.6(C-5), 41.5(C-10), 41.4(C- 9), 39.8(C-8), 38.2(C-22), 37.2(C-14), 36.4(C-4), 32.3(C-7), 31.8(C-17), 31.3(C-15 ),28.5(C-27),28.1(C-28),26.8(C-16),26.0(C-21),25.9(C-23),25.8(C-29),23.8(C-24) ,23.6(C-11),17.3(C-6),16.9(C-25),16.4(C-26); EI(m/z):558(M ⁺ -2H),247(100),542,451,423,405,338,247 ,91,69,43.

Compound Iw, white powder, C ₃₇ H ₅₂ O ₄ , m.p79～81℃; ¹ H NMR (400MHz, CDCl ₃ )δ: 7.35-7.27(m,5H,H-Ar), 5.16(d,1H, J=12.4Hz, Bn-CHH'),5.15(brs,1H,H-12),5.06(d,1H,J=12.4Hz,Bn-CHH'),3.48(brs,1H,H-3), 3.32(t,1H,J=4.0Hz,H-2),3.16(d,1H,J=4.0Hz,H-1),1.19(s,3H),1.15(s,3H),1.00(s, 3H),0.94(s,3H),0.77(s,3H),0.78(s,3H),0.69(s,3H); EI(m/z):560(M ⁺ ),247(100),451,423,338,247 ,91,69,43.

Compound Ix, white powder, C ₃₇ H ₅₂ O ₄ , m.p113～115℃; ¹ H NMR (400MHz, CDCl ₃ )δ: 7.33-7.29(m,5H,H-Ar), 5.18(d,1H, J=12.4Hz, Bn-CHH'),5.17(brs,1H,H-12),5.04(d,1H,J=12.4Hz,Bn-CHH'),3.47(d,1H,J=3.6Hz, H-3),3.40(brs,1H,H-2),3.15(d,1H,J＝4.0Hz,H-1),1.16(s,3H),1.10(s,3H),1.07(s, 3H),0.98(s,3H),0.90(s,3H),0.88(s,3H),0.75(s,3H); EI(m/z):560(M ⁺ ),247(100),542,451,423,405,338,247 ,91,69,43.

Embodiment 11: the preparation of compound Ie, If, Ig, Ih:

Compounds Ie, If, Ig, and Ih were prepared by using 0.11 mmol of compounds Ii, Ij, Ik, and Il as raw materials respectively, adding 100 ml of methanol to dissolve them, adding 18 mg of palladium carbon, introducing hydrogen, stirring the reaction at room temperature, and detecting the reaction by TLC. process, the reaction was completed in 10 hours, the palladium carbon was recovered by filtration, the filtrate was concentrated under reduced pressure to obtain the crude product, and the pure product was obtained by silica gel column chromatography.

Compound Ie, yield 81.5%, white powder, C ₃₀ H ₄₄ O ₅ , m.p 156～158°C; ¹ H NMR (400MHz, CDCl ₃ )δ: 5.20(s, 1H, H-12), 3.40(d ,1H,J=4.0Hz,H-3),3.35(t,1H,J=4.8Hz,H-2),3.20(d,1H,J=4.0Hz,H-1),1.25(s,3H ),1.19(s,3H),1.09(s,3H),0.98(s,3H),0.87(s,3H),0.78(s,3H),0.72(s,3H); EI(m/z) :484(M ⁺ ),247(100),466,469,338,247.

Compound If, yield 37.2%, white powder, C ₃₀ H ₄₄ O ₅ , m.p97～99℃; ¹ H NMR (400MHz, CDCl ₃ )δ: 5.20(s, 1H, H-12), 3.45(d ,1H,J=4.4Hz,H-3),3.38(brs,1H,H-2),3.16(d,1H,J=4.0Hz,H-1),1.20(s,3H),1.16(s ,3H),1.12(s,3H),0.96(s,3H),0.93(s,3H),0.85(s,3H),0.75(s,3H); EI(m/z):484(M ⁺ ), 247(100), 466, 469, 338, 247.

Compound Ig, white powder, C ₃₀ H ₄₆ O ₄ , m.p135～137°C; ¹ H NMR (400MHz, CDCl ₃ ) δ: 5.18(brs,1H,H-12), 3.59(d,1H,J= 3.6Hz, H-3), 3.07(brs, 1H, H-2), 2.93(d, 1H, J=4.0Hz, H-1), 1.25(s, 3H), 1.19(s, 3H), 1.14 (s,3H),0.99(s,3H),0.94(s,3H),0.79(s,3H),0.75(s,3H); EI(m/z):470(M ⁺ ),247(100 ), 452, 435, 338, 247, 138.

Compound Ⅰh, yield 82.4% white powder, C ₃₀ H ₄₆ O ₄ , m.p130～132°C; ¹ H NMR (400MHz, CDCl ₃ ) 5.25(s,1H,H-12), 4.19(d,1H, J＝4.0Hz, H-3), 3.64(t, 1H, J＝4.4Hz, H-2), 3.32(t, 1H, J＝4.4Hz, H-1), 2.90(s, 1H, H- 9),1.50(s,3H),1.48(s,3H),1.27(s,3H),1.15(s,3H),0.81(s,3H),0.82(s,3H),0.58(s,3H ); EI (m/z): 470 (M ⁺ ), 247 (100), 452, 435, 138.

Embodiment 12: the preparation of compound Iq, Ir, Is, It:

Compounds Iq, Ir, Is, Itt are prepared from compounds Iu, Iv, Iw, Ix respectively, and the method is the same as that of compounds Ie, If, Ig, Ih.

Compound Iq, yield 24.6%, white powder, C ₃₀ H ₄₄ O ₅ , m.p98～100℃; ¹ H NMR (400MHz, CDCl ₃ )δ: 5.20(s,1H,H-12), 4.04(d ,1H,J=4.0Hz,H-3),3.86(brs,1H,H-2),3.13(d,1H,J=4.0Hz,H-1),2.24(s,3H),2.04(s ,3H),1.17(s,3H),0.96(s,3H),0.93(s,3H),0.88(s,3H),0.79(s,3H); EI(m/z):484(M ⁺ ), 247(100), 466, 469, 338, 247.

Compound Ⅰr, yield 18.6%, white powder, C ₃₀ H ₄₄ O ₅ , m.p93～95°C; ¹ H NMR (400MHz, CDCl ₃ ) δ: 5.22(s, 1H, H-12), 4.00(d ,1H,J=2.8Hz,H-3),3.96(brs,1H,H-2),3.82(d,1H,J=4.0Hz,H-1),2.13(s,3H),2.00(s ,3H),1.18(s,3H),0.98(s,3H),0.93(s,3H),0.78(s,3H),0.73(s,3H); EI(m/z):484(M ⁺ ), 247(100), 466, 469, 338, 247, 43.

Compound Is, yield 38.4% white powder, C ₃₀ H ₄₆ O ₄ , m.p157～159°C; ¹ H NMR (400MHz, CDCl ₃ ) 5.22(s, 1H, H-12), 4.10(d, 1H, J＝4.4Hz, H-3), 3.85(t, 1H, J＝4.4Hz, H-2), 3.51(d, 1H, J＝4.4Hz, H-1), 2.90(s, 1H, H- 9),1.94(s,3H),1.56(s,3H),1.21(s,3H),1.10(s,3H),0.91(s,3H),0.85(s,3H),0.68(s,3H ); EI (m/z): 470 (M ⁺ ), 247 (100), 452, 435, 138.

Compound It, yield 22.7% white powder, C ₃₀ H ₄₆ O ₄ , m.p101～103°C; ¹ H NMR (400MHz, CDCl ₃ ) 5.24(s, 1H, H-12), 4.09(d, 1H, J＝3.6Hz, H-3), 3.88(brs, 1H, H-2), 3.68(d, 1H, J＝4.4Hz, H-1), 2.90(s, 1H, H-9), 1.55( s,3H),1.41(s,3H),1.37(s,3H),1.19(s,3H),0.89(s,3H),0.82(s,3H),0.60(s,3H); EI(m /z): 470(M ⁺ ), 247(100), 452, 435, 138.

Embodiment 13: the preparation of compound Ia, Im:

Compounds Ia and Im were prepared using compounds Ic and Io as raw materials respectively, and the methods were the same as those for compounds Ie, If, Ig and Ih.

Compound Ia, white powder, C ₃₀ H ₄₂ O ₅ , m.p169～171°C; ¹ H NMR (400MHz, CDCl ₃ )δ: 5.60(s, 1H, H-12), 4.49(d, 1H, J＝ 4.4Hz, H-2), 4.18(d, 1H, J=4.4Hz, H-1), 2.90(s, 1H, H-9), 1.65(s, 3H), 1.44(s, 3H), 1.28 (s,3H),1.17(s,3H),1.10(s,3H),0.89(s,3H),0.72(s,3H); EI(m/z):482(M ⁺ ),467,247(100 ), 202, 138, 43.

Compound Im, white powder, C ₃₀ H ₄₂ O ₅ , m.p128～130°C; ¹ H NMR (400MHz, CDCl ₃ )δ: 5.60(s, 1H, H-12), 4.50(d, 1H, J= 4.0Hz, H-2), 4.47(d, 1H, J=4.0Hz, H-1), 2.91(s, 1H, H-9), 1.51(s, 3H), 1.28(s, 3H), 1.26 (s,3H),1.13(s,3H),1.03(s,3H),0.81(s,3H),0.77(s,3H); EI(m/z):482(M ⁺ ),467,247(100 ), 202, 138, 43.

Embodiment 14: the preparation of compound Ib, In:

Compounds Ib and In were prepared using compounds Id and Ip as raw materials respectively, and the method was the same as that of compounds Ie, If, Ig and Ih.

Compound Ib, white powder, C ₃₀ H ₄₄ O ₄ , m.p84～86°C; ¹ H NMR (400MHz, CDCl ₃ )δ: 5.30(s, 1H, H-12), 3.57(d, 1H, J= 4.8Hz, H-2), 3.39(d, 1H, J=4.8Hz, H-1), 2.92(s, 1H, H-9), 1.25(s, 3H), 1.20(s, 3H), 1.15 (s,3H),0.96(s,3H),0.84(s,3H),0.81(s,3H),0.72(s,3H); EI(m/z):468(M ⁺ ),422,248,202(100 ), 202, 43.

Compound In, white powder, C ₃₀ H ₄₄ O ₄ , m.p 114～116°C; ¹ H NMR (400MHz, CDCl ₃ )δ: 5.30(s, 1H, H-12), 3.59(d, 1H, J= 4.8Hz, H-2), 3.43(d, 1H, J=4.8Hz, H-1), 2.92(s, 1H, H-9), 1.21(s, 3H), 1.20(s, 3H), 1.18 (s,3H),0.99(s,3H),0.85(s,3H),0.79(s,3H),0.70(s,3H); EI(m/z):468(M ⁺ ),422,248,202(100 ), 202, 43.

Embodiment 15: Preparation of benzyl substituted ring-opened compound:

Take 200.00mg (0.35mmol) of compound Ii, Ij, Iu, Iv or Ik, Il, Iw, Ix dissolved in 50ml of acetone, stir in ice bath for 15min, then add perchloric acid 25μL (0.45mmol) dropwise, add distilled water 10μL, nitrogen Stir at room temperature under protection until the reaction is complete (TLC detects the progress of the reaction). After 15 hours of reaction completion, adjust the pH value of the reaction solution to 7 with 2mol/L sodium hydroxide solution, evaporate the acetone under reduced pressure, add water and ethyl acetate to the residue for extraction, and wash the organic layer with saturated _NaHCO3 solution and saturated NaCl solution in turn and anhydrous magnesium sulfate, dried and filtered, and the solvent was distilled off under reduced pressure to obtain a crude white powder. Perform silica gel column chromatography (V (petroleum ether): V (acetone) = 2:1) to obtain benzyl-substituted ring-opened compounds, such as G-6-2, G-6-4, and Ggc.

Compound G-6-2, yield 73.5%, white powder, C ₃₇ H ₅₂ O ₆ , m.p89～91℃.IR(KBr),σ/cm ^-1 :3441,2948,2863,1731,1655, 1455, 1384, 1214, 1159, 1045, 999, 732, 695; ¹ H NMR (400Hz, CDCl ₃ ) δ: 7.41-7.33 (m, 5H, H-Ar), 5.61 (s, 1H, H-12), 5.20 (d ,1H,J=12.4Hz,Bn-CHH'),5.09(d,1H,J=12.0Hz,Bn-CHH'),4.69(brs,1H,H-1),4.00(brs,1H,H- 2),3.57(brs,1H,H-3),3,28(brs,1H,OH),3.24(s,1H,H-9),3.00(brs,1H,OH),2.68(brs,1H ,OH),1.39(s,3H),1.36(s,3H),1.16(s,3H),1.15(s,3H),1.02(s,3H),0.99(s,3H),0.74(s, 3H); ¹³ C NMR (100MHz, CDCl ₃ ) δ: 202.7(C-11), 176.3(C-30), 171.4(C-13), 136.1(C _ar ), 128.6(C _ar ), 128.6(C _ar ),128.4(C-12),128.3(C _ar ),128.2(C _ar ),128.2(C _ar ),76.0(C-1),75.2(C-2),74.5(C-3),66.2 (Bn-CH ₂ ), 54.7(C-9), 48.0(C-18), 47.5(C-5), 44.9(C-10), 43.9(C-20), 43.6(C-14), 41.2 (C-8), 41.1(C-19), 38.0(C-4), 37.7(C-22), 31.8(C-21), 31.7(C-17), 31.1(C-7), 29.8( C-23), 28.5(C-28), 28.2(C-29), 26.5(C-15), 26.4(C-16), 23.5(C-27), 19.2(C-26), 17.7(C -6, C-24), 16.9 (C-25); EI (m/z): 592 (M ⁺ ), 91 (100), 574, 557, 541, 491, 393, 352, 225, 135, 69, 43.

Compound G-6-4, yield 46.8%, white powder, C ₃₇ H ₅₂ O ₆ , m.p114～116℃.IR(KBr),σ/cm ^-1 :3424,2959,2869,1728,1650, 1456,1385,1214,1152,1007,876,751,698; ¹ H NMR (400MHz, CDCl ₃ )δ:7.41-7.32(m,5H,H-Ar),5.70(s,1H,H-12),5.21(d ,1H,J=12.0Hz,Bn-CHH'),5.10(d,1H,J=12.4Hz,Bn-CHH'),3.90(dd,1H,J=4.0Hz,7.2Hz,H-1), 3.72(d,1H,J=11.6Hz,H-3),3.54(dd,1H,J=8.0Hz,11.6Hz,H-2),3.14(s,1H,H-9),2.89(brs, 1H,OH),2.77(brs,1H,OH),2.48(brs,1H,OH),1.35(s,3H),1.17(s,3H),1.16(s,3H),1.15(s,3H) ,1.02(s,3H),0.94(s,3H),0.76(s,3H); ¹³ C NMR(100MHz,CDCl ₃ )δ:203.6(C-11),176.1(C-30),172.5(C -13),136.0(C _ar ),128.5(C _ar ),128.5(C _ar ),128.3(C _ar ),128.2(C-12),128.1(C _ar ),128.1(C _ar ),82.6(C ar -1), 74.6(C-2), 74.3(C-3), 66.2(Bn-CH ₂ ), 54.5(C-9), 48.1(C-18), 46.8(C-5), 44.5(C -10), 43.9(C-20), 43.7(C-14), 42.0(C-8), 41.1(C-19), 37.6(C-22), 37.5(C-4), 31.6(C- 17), 31.0(C-21), 30.9(C-7), 28.5(C-29), 28.1(C-28), 26.5(C-15), 26.3(C-16), 23.4(C-27 ),23.3(C-23,C-24),21.3(C-25),19.3(C-6),18.8(C-26); EI(m/z):574(M ⁺ -H ₂ O) ,91(100),,557,541,491,393,352,225,135,69,43.

Compound Ggc, yield 78.8%, white powder, C ₃₇ H ₅₄ O ₅ , _m.p 110～112℃; IR(KBr),σ/cm ^-1 :3425,2949,1729,1455,1382,1214,1155 , 1029,697; ¹ H NMR (400MHz, CDCl ₃ ) δ: 7.39-7.32 (m, 5H, H-Ar), 5.19 (d, 1H, Bn-CHH', J=12.0Hz), 5.13 (brs, 1H,H-12),5.07(d,1H,Bn-CHH',J=12.0Hz),3.93(t,1H,J=4.0Hz,H-2),3.61(d,J=2.4Hz,1H ,H-3),3.45(d,1H,J=4.0Hz,H-1),1.24(s,3H),1.17(s,3H),1.11(s,3H),1.00(s,3H), 0.99(s,3H),0.97(s,3H),0.71(s,3H); ¹³ C NMR(100MHz,CDCl ₃ )δ:177.1(C-30),144.5(C-13),136.3(C _ar ),128.4(C _ar ),128.4(C _ar ),128.0(C _ar ),128.0(C _ar ),128.0(C _ar ),122.2(C-12),75.9(C-1),74.5(C- 2), 73.9(C-3), 65.9(Bn-CH ₂ ), 48.1(C-18), 47.9(C-5), 44.2(C-20), 42.7(C-19), 42.0(C- 10), 40.2(C-4), 39.6(C-8), 38.5(C-22), 38.3(C-9), 38.2(C-14), 32.0(C-7), 31.8(C-17 ), 31.2(C-15), 29.5(C-27), 28.5(C-23), 28.1(C-28), 26.9(C-16), 26.0(C-21), 26.0(C-29) ,23.4(C-11),17.9(C-6),17.2(C-24),17.0(C-25),14.3(C-26); EI(m/z):578(M ⁺ ),247 (100), 560, 542, 527, 423, 405, 338, 247, 91, 69, 43.

Example 16: Preparation of Compounds IIa～IIh, IIi～IIp:

Take 100.00 mg (0.17 mmol) of the benzyl-substituted ring-opened compound prepared in Example 15 and dissolve it in 50 ml of methanol, add 20 mg of palladium carbon, feed hydrogen, and stir at room temperature for the reaction. TLC detects the reaction process (V (petroleum ether): V ( Acetone)=3:1), the reaction was completed in 12h, the palladium carbon was recovered by filtration, the filtrate was concentrated under reduced pressure to obtain the crude product, and the corresponding debenzyl ring-opened compound IIa～IIh or IIi～IIp was obtained by silica gel column chromatography.

Compound Ⅱa, yield 83.0%, white powder, m.p113～115℃; IR(KBr),σ/cm ^-1 :3443,2925,1627,1384; ¹ H-NMR(CD ₃ OD)δ:5.24( s,1H,H-12),3.93(d,1H,J=6.8Hz,H-2),3.61(d,1H,J=2.4Hz,H-3),3.45(d,1H,J=4.4 Hz, H-1); ¹³ C-NMR (100MHz, CD ₃ OD) δ: 205.9 (C-11), 180.5 (C-30), 139.5 (C-13), 127.1 (C-12), 77.4 ( C-3), 75.3(C-1), 74.9(C-2), 49.6(C-5), 47.6(C-17), 47.2(C-19), 43.4(C-14), 42.7(C -18), 41.5(C-8), 40.3(C-4), 39.7(C-9), 39.3(C-20), 39.2(C-10), 34.8(C-21), 33.8(C- 7), 33.6(C-29), 33.5(C-22), 30.1(C-30), 28.8(C-15), 26.5(C-27), 24.4(C-16), 24.0(C-28 ),24.0(C-11),19.1(C-6),18.1(C-25),17.8(C-24),14.8(C-26); EI(m/z):502(M ⁺ ), 91 (100), 484, 469, 401, 303, 207, 135, 43.

Compound Ⅱb, yield 80.5%, white powder, m.p167～169℃; IR(KBr),σ/cm ^-1 :3431,2949,2867,1704,1646,1464,1387,1219,1117,1049,975,819 ; ¹ H NMR (400MHz, C ₅ D ₅ N) δ: 6.75(brs,1H,OH),6.06(s,1H,H-12),5.74(brs,1H,H-1),4.81(t, 1H,J=3.6Hz,H-2),4.28(d,1H,J=4.0Hz,H-3),3.63(s,1H,H-9),2.56(d,1H,J=10.4Hz, OH),2.29(d,1H,J=11.2Hz,OH),1.78(s,3H),1.51(s,3H),1.46(s,3H),1.43(s,3H),1.32(s,3H ),1.29(s,3H),0.83(s,3H); ¹³ C NMR(100MHz,C ₅ D ₅ N)δ:201.4(C-11),182.9(C-30),169.7(C-13) ,129.0(C-12),77.4(C-2),74.8(C-1),74.4(C-3),54.9(C-9),48.7(C-18),48.4(C-5), 45.4(C-14), 44.2(C-10), 44.1(C-8), 41.9(C-20), 41.7(C-19), 39.6(C-4), 38.5(C-22), 32.8 (C-7), 32.2(C-17), 31.6(C-21), 30.4(C-23), 28.8(C-29), 28.8(C-28), 26.8(C-15), 26.7( C-16),23.7(C-27),19.5(C-26),18.1(C-24),17.8(C-6),16.6(C-25).EI(m/z):503(M ⁺ +1),91(100),484,469,401,303,207,135,43.

Compound Ⅱc, yield 32.5%, white powder, m.p136～138℃; ¹ H NMR (400MHz, C ₅ D ₅ N) δ: 6.06(s,1H,H-12), 5.66(brs,1H,H -1),4.95(dd,1H,J=3.6Hz,11.6Hz,H-2),4.34(d,1H,J=4.0Hz,H-3),3.62(s,1H,H-9), 2.23(d,1H,J=10.4Hz,OH),2.14(d,1H,J=11.2Hz,OH),1.76(s,3H),1.52(s,3H),1.40(s,3H),1.38 (s, 3H), 1.35 (s, 3H), 1.24 (s, 3H), 0.81 (s, 3H); EI (m/z): 502 (M ⁺ ), 91 (100), 484, 469, 303, 207, 135, 43.

Compound Ⅱd, yield 22.8%, white powder, m.p145～147℃; ¹ H NMR (400MHz, C ₅ D ₅ N) δ: 6.06(s, 1H, H-12), 5.55(d, 1H, J =4.4Hz,H-1),5.04(d,1H,J=4.0Hz,H-2),4.74(d,1H,J=3.6Hz,H-3),3.63(s,1H,H-9 ),1.84(s,3H),1.76(s,3H),1.58(s,3H),1.34(s,3H),1.22(s,3H),1.21(s,3H),0.95(s,3H) ; EI (m/z): 502 (M ⁺ ), 91 (100), 484, 469, 303, 207, 135, 43.

Compound Ⅱe, yield 79.5%, white powder, m.p125～127℃; ¹ H-NMR (400MHz, CD ₃ OD) δ: 5.24(s, 1H, H-12), 3.93(d, 1H, J= 6.8Hz, H-1), 3.61(d, 1H, J=2.4Hz, H-3), 3.45(d, 1H, J=4.4Hz, H-2), 1.28, 1.23, 1.13, 1.00, 0.87( 3H×7, s, 23, 24, 25, 26, 27, 28, 29); EI (m/z): 502 (M ⁺ ), 91 (100), 484, 469, 401, 303, 207, 135, 43.

Compound IIf, yield 85.6%, white powder, m.p 155-157°C; 502(M ⁺ ), 91(100), 484, 401, 303, 207, 135, 43.

Compound IIg, yield 81.5%, white powder, m.p199～201°C; 503(M ⁺ +1), 91(100), 484, 469, 401, 303, 207, 135.

Compound Ⅱh, yield 18.7%, white powder, m.p185～187℃; ¹ H NMR (400MHz, C ₅ D ₅ N) δ: 6.08(s, 1H, H-12), 5.68(brs, 1H, H -1),4.99(brs,H-2),4.50(d,1H,J=11.6Hz,H-3),3.62(s,1H,H-9),2.00(d,1H,J=10.4Hz ,OH),1.95(d,1H,J=11.2Hz,OH),1.85(s,3H),1.63(s,3H),1.58(s,3H),1.40(s,3H),1.30(s, 3H), 1.14 (s, 3H), 0.86 (s, 3H); EI (m/z): 502 (M ⁺ ), 91 (100), 484, 469, 303, 207, 135, 43.

Compound Ⅱi, yield 85.5%, white powder, C ₃₀ H ₄₈ O ₅ , m.p 194～196℃; IR(KBr),σ/cm ^-1 :1695,3443,2947,1643,1386; ¹ H-NMR (400MHz,CD ₃ OD)δ:5.24(s,1H,H-12),3.91(d,1H,J＝6.8Hz,H-2),3.59(d,1H,J＝2.8Hz,H-3 ),3.44(d,1H,J＝10.0Hz,H-1),1.20,1.18,1.04,0.99,0.92,0.88,0.83(3H×7,s,23,24,25,26,27,28, 29); ¹³ C-NMR (100MHz, CD ₃ OD) δ: 180.5 (C-30), 144.9 (C-13), 123.9 (C-12), 77.4 (C-3), 75.3 (C-1) ,74.9(C-2),49.6(C-5),47.6(C-17),47.2(C-19),43.4(C-14),42.7(C-18),41.5(C-8), 40.3(C-4), 39.7(C-9), 39.3(C-20), 39.2(C-10), 34.8(C-21), 33.8(C-7), 33.6(C-29), 33.5 (C-22), 30.1 (C-30), 28.8 (C-15), 26.5 (C-27), 24.4 (C-16), 24.0 (C-28), 24.0 (C-11), 19.1 ( C-6), 18.1 (C-25), 17.8 (C-24), 14.8 (C-26); MS (EI) m/z: 488 (M ⁺ ), 203 (100), 248, 119, 133, 452, 470.

Compound Ⅱj, yield 89.0%, white powder, C ₃₀ H ₄₈ O ₅ , m.p 255～257°C; IR(KBr), σ/cm ^-1 :3428, 2948, 1701, 1455, 1383, 1227, 1177, 1029,969; ¹ H NMR (400MHz, C ₅ D ₅ N) δ: 6.35 (brs, 1H), 5.59 (brs, 1H, H-12), 4.80 (t, 1H, J=4.0Hz, H-2 ),4.29(brs,1H,H-1),4.27(d,1H,J=4.0Hz,H-3),3.03(q,J=4.0Hz,1H,H-9),1.69(s,3H ),1.49(s,3H),1.41(s,3H),1.40(s,3H),1.40(s,3H),1.18(s,3H),0.95(s,3H); ¹³ C NMR(100MHz, C ₅ D ₅ N)δ: 179.8(C-30), 145.1(C-13), 123.5(C-12), 77.6(C-2), 74.7(C-3), 74.3(C-1), 49.1(C-5), 48.8(C-18), 44.4(C-20), 43.6(C-19), 42.7(C-8), 41.2(C-10), 40.1(C-14), 39.3 (C-4), 39.2(C-22), 38.9(C-9), 32.9(C-7), 32.5(C-17), 31.9(C-21), 30.5(C-23), 29.2( C-29), 28.7(C-28), 27.4(C-16), 26.7(C-15), 26.4(C-27), 24.0(C-11), 18.7(C-6), 18.3(C -24), 17.6 (C-26), 15.0 (C-25); EI (m/z): 488 (M ⁺ ), 248 (100), 470, 452, 437, 233, 98, 43.

Compound Ⅱk, yield 56.4%, white powder, C ₃₀ H ₄₈ O ₅ , m.p148～150℃; ¹ H NMR (400MHz, C ₅ D ₅ N) δ: 5.60(brs,1H,H-12), 4.87(t,1H,J=4.0Hz,H-2),4.46(d,1H,J=4.4Hz,H-1),4.38(d,1H,J=4.0Hz,H-3),3.03( q,J＝4.0Hz,1H,H-9),1.66(s,3H),1.44(s,3H),1.41(s,3H),1.35(s,3H),1.34(s,3H),1.12 (s,3H), 0.97(s,3H); MS (EI) m/z: 488 (M ⁺ ), 203 (100), 248, 119, 133, 452, 470.

Compound IIl, yield 32.0%, white powder, C ₃₀ H ₄₈ O ₅ , m.p122～124°C; ¹ H NMR (400MHz, C ₅ D ₅ N) δ: 5.59(brs,1H,H-12), 4.92(dd, 1H, J=4.0Hz, 11.6Hz, H-2), 4.54(d, 1H, J=11.6Hz, H-1), 4.36(d, 1H, J=4.4Hz, H-3) ,3.04(q,J＝4.0Hz,1H,H-9),1.75(s,3H),1.53(s,3H),1.46(s,3H),1.42(s,3H),1.40(s,3H ), 1.08(s,3H), 0.98(s,3H); MS(EI) m/z: 488(M ⁺ ), 203(100), 248,119,133,452,470.

Compound Ⅱm, yield 73.1%, white powder, C ₃₀ H ₄₈ O ₅ , m.p183～185℃; ¹ H NMR (400MHz, C ₅ D ₅ N) δ: 5.59(brs,1H,H-12), 4.95(brs,1H,H-2),4.51(d,1H,J=4.4Hz,H-1),4.31(d,1H,J=4.4Hz,H-3),3.03(s,1H,H -9),1.72(s,3H),1.69(s,3H),1.51(s,3H),1.45(s,3H),1.42(s,3H),1.28(s,3H),0.99(s, 3H); MS (EI) m/z: 488 (M ⁺ ), 203 (100), 248, 119, 133, 452, 470.

Compound IIn, yield 75.7%, white powder, C ₃₀ H ₄₈ O ₅ , m.p 238～240°C; EI(m/z): 488(M ⁺ ), 248(100), 470, 452, 437, 233, 98, 43.

Compound IIo, yield 78.9%, white powder, C ₃₀ H ₄₈ O ₅ , m.p 178～180°C; EI(m/z): 488(M ⁺ ), 248(100), 470, 452, 437, 233, 98, 43.

Compound Ⅱp, yield 9.8%, white powder, C ₃₀ H ₄₈ O ₅ , m.p168～170℃; I ¹ H NMR (400MHz, C ₅ D ₅ N) δ: 5.61(brs,1H,H-12) ,4.85(brs,1H,H-2),4.72(d,1H,J=4.4Hz,H-1),4.23(d,1H,J=11.6Hz,H-3),3.03(s,1H, H-9),1.69(s,3H),1.55(s,3H),1.48(s,3H),1.31(s,3H),1.14(s,3H),1.12(s,3H),0.98(s , 3H); MS (EI) m/z: 488 (M+), 203 (100), 248, 119, 133, 452, 470.

Example 17: Preparation of compounds in which R ₄ is -COOR ¹ in general formulas I and II:

Take 1 mmol of the compound whose R ₄ is -COOH in the general formula I or II as the raw material, dissolve it in N,N-dimethylformamide (DMF), add 5 mmol of potassium carbonate and stir for 30 minutes, then add the haloalkane (R ¹ X) 1.1 mmol, reacted at room temperature under nitrogen protection, and detected the progress of the reaction by TLC. After the reaction is complete, water and ethyl acetate are added for extraction, the ethyl acetate layer is dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain a crude product, which is separated and purified by silica gel column chromatography to obtain the compound in which R ₄ is -COOR ¹ in the general formula I (such as GFO-715) or compounds in which R ₄ is -COOR ¹ in the general formula II (such as G-8, GF-73, GLP-h).

Compound G-8, yield 95.5%, white powder, C ₃₁ H ₄₈ O ₆ , m.p 148～150°C; IR(KBr), σ/cm ^-1 :3438, 2945, 2864, 1733, 1655, 1464, 1386,1216,1155,998,866; ¹ H NMR (400MHz, CDCl ₃ ) δ: 5.72(s,1H,H-12),4.68(brs,1H,H-1),4.00(brs,1H,H-2 ),3.70(s,3H,OCH ₃ ),3.57(brs,1H,H-3),3.29(brs,1H,OH),3.24(s,1H,H-9),3.03(brs,1H,OH ),2.69(brs,1H,OH),1.41(s,3H,H-25),1.39(s,3H,H-27),1.19(s,3H,H-29),1.18(s,3H, H-26),1.02(s,3H,H-23),0.99(s,3H,H-24),0.88(s,3H,H-28); ¹³ C NMR(100MHz,CDCl ₃ )δ:202.7 (C-11),177.0(C-30),171.4(C-13),128.4(C-12),76.0(C-1),75.3(C-2),74.3(C-3),54.7( C-9), 51.8(OCH ₃ ), 48.2(C-18), 47.5(C-5), 44.9(C-10), 44.0(C-20), 43.7(C-14), 41.2(C- 8), 41.1(C-19), 38.0(C-4), 37.7(C-22), 31.8(C-21), 31.7(C-17), 31.1(C-7), 29.8(C-23 ),28.5(C-28),28.2(C-29),26.5(C-15),26.4(C-16),23.5(C-27),19.2(C-26),17.7(C-6, C-24), 16.9 (C-25); EI (m/z): 516 (M+), 498 (100), 483, 415, 385, 317, 276, 135.

Compound GF-73, yield 96.0%, light yellow powder, C ₃₇ H ₅₂ O ₆ , m.p89～91℃; IR(KBr), σ/cm ^-1 :3425, 2944, 2870, 1727, 1652, 1460 , 1384, 1216, 1173, 1082, 1033, 817, 768; ¹ H NMR (400MHz, CDCl ₃ ) δ: 5.72 (s, 1H, H-12), 4.61 (brs, 1H, H-1), 3.97 (brs, 1H,H-2),3.42(brs,1H,H-3),3.21(s,1H,H-9),2.85(brs,1H,OH),2.64(brs,1H,OH),2.45(brs ,1H,OH),1.38(s,3H,H-25),1.37(s,3H,H-27),1.18(s,3H,H-29),1.14(s,3H,H-26), 1.02(s,3H,H-23),0.99(s,3H,H-24),0.83(s,3H,H-28); ¹³ C NMR(100MHz,CDCl ₃ )δ:203.0(C-11) ,176.5(C-30),172.0(C-13),128.5(C-12),76.0(C-1),75.2(C-2),74.7(C-3),60.35(COOCH ₂ ),54.7 (C-9), 48.2(C-18), 47.6(C-5), 44.9(C-10), 43.8(C-20), 43.7(C-14), 41.3(C-8), 41.2( C-19), 37.9(C-4), 37.8(C-22), 31.8(C-21), 31.7(C-17), 31.1(C-7), 29.9(C-23), 28.7(C -28),28.2(C-29),26.5(C-15),26.4(C-16),23.6(C-27),19.2(C-26),17.8(C-6)),17.0(C -24), 14.3 (C-25); ESI (m/z): 532 (M+), 532 (M+-H).

Compound GLP-h, yield 93.5%, white powder, C ₃₇ H ₅₃ ClO ₅ , m.p98～100℃; IR(KBr),σ/cm ^-1 :3383,2939,1728,1640,1457,1383, 1158,1028,803,767,672; ¹ H NMR (400MHz, CDCl ₃ )δ:7.34(m,4H,Ph),5.60(brs,1H,H-12),5.12(d,1H,J＝12.0Hz,Bn- CHH'), 5.09 (d, 1H, J=12.4Hz, Bn-CHH'), 3.90 (brs, 1H, H-2), 3.50 (brs, 1H, H-1), 3.18 (s, 1H, H -3),2.56(brs,1H,OH),2.45(brs,1H,OH),2.28(brs,1H,OH),1.37(s,3H),1.35(s,3H),1.17(s,3H ),1.14(s,3H),1.02(s,3H),0.99(s,3H),0.77(s,3H); EI(m/z):612(M ⁺ ),256(100),522,488,358,437,233,126, 43.

Compound GFO-715, yield 96.8%, white powder, C ₄₅ H ₇₂ O ₅ , m.p78～80℃; IR(KBr),σ/cm ^-1 :3424,2923,2852,1728,1656,1463, 1385, 1257, 1213, 1116, 1044, 722; ¹ H NMR (400MHz, CDCl ₃ ) δ: 5.60 (s, 1H, H-12), 4.45 (d, 1H, J=4.4Hz, H-2), 4.09(t,2H,COOCH ₂ ),3.34(d,1H,J＝4.4Hz,H-1),2.98(s,1H,H-9),1.38(s,3H),1.37(s,3H) ,1.18(s,3H),1.14(s,3H),1.02(s,3H),0.99(s,3H),0.89(t,3H),0.83(s,3H); ¹³ C NMR (100MHz, CDCl ₃ ) δ: 203.0(C-3), 197.3(C-11), 176.0(C-30), 170.1(C-13), 127.1(C-12), 64.6(COO-CH ₂ ), 62.6(C -1), 56.4(C-2), 54.5(C-9), 48.0(C-18), 47.5(C-5), 45.6(C-4), 43.8(C-14), 43.1(C- 20), 41.6 (C-10), 41.0 (C-19), 36.3 (C-8), 37.1 (C-22), 31.1 (C-7), 31.0 (C-17), 29.7, 29.6, 29.5, 29.5, 29.5, 29.4, 29.3, 29.1, 28.7, 28.7, 28.6, 28.3, 28.2(13×CH ₂ ), 28.5(C-21), 28.3(C-29), 28.2(C-28), 27.9(C -23), 26.1(C-15), 24.2(C-16), 23.1(C-27), 20.3(C-24), 17.6(C-25), 17.1(C-6), 14.8(C- 26); EI (m/z): 692 (M ⁺ ), 677, 457 (100), 472, 224.

Example 18: Preparation of compounds in which R4 is -CONHR ¹ or -CONR ¹ R ² in general formulas I and II:

Take 1 mmol of the compound whose R ₄ is -COOH in the general formula I or II as the raw material, dissolve it in N,N-dimethylformamide (DMF), add 1.5 mmol of R ¹ NH ₂ or R ¹ R ² NH, and then add 3 mmol of N, N'-dicyclohexylcarbodiimide (DCC) and 0.1 mmol of 4-dimethylaminopyridine (DMAP) were reacted at room temperature under the protection of nitrogen, and the reaction progress was detected by TLC. After the reaction is complete, add 2mol/L hydrochloric acid solution to terminate the reaction, add water and ethyl acetate to extract, the ethyl acetate layer is dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain a crude product, which is separated and purified by silica gel column chromatography to obtain the general formula Ⅰ A compound in which R ₄ is -CONHR ¹ or -CONR ¹ R ² (such as GO-h), or a compound in which R ₄ is -CONHR ¹ or -CONR ¹ R ² in general formula II (such as GN-g1).

Compound GN-g1, yield 90.3%, white powder, C ₃₁ H ₅₁ NO ₄ , m.p146～148°C; ¹ H NMR (400MHz, CDCl ₃ ) δ: 5.20(s, 1H, H-12), 3.70 (d,1H,J=4.0Hz,H-1),3.72(d,1H,J=4.0Hz,H-3),3.64(d,1H,J=3.6Hz,11.6Hz,H-2), 2.69(brs,1H,OH),2.57(brs,1H,OH),2.49(brs,1H,OH),2.37(q,1H,NH),1.47(s,3H,NHCH ₃ ),1.30(s, 3H,H-27),1.15(s,3H,H-29),1.14(s,3H,H-25),1.12(s,3H,H-26),1.07(s,3H,H-23) , 0.84 (s, 3H, H-24), 0.70 (s, 3H, H-28); EI (m/z): 501 (M ⁺ ), 247 (100), 483, 261, 188, 58.

Compound GO-h, yield 91.5%, white powder, C ₃₇ H ₅₁ NO ₃ , m.p123～125°C; ¹ H NMR (400MHz, CDCl ₃ ) δ: 7.27-7.23 (m, 5H, H-Ar) ,5.22(brs,1H,H-12),3.20(s,1H,NH),3.46(d,J=4.4Hz,1H,H-1),3.32(d,J=4.4Hz,1H,H- 2),1.30(s,3H,H-27),1.26(s,3H,H-29),1.19(s,3H,H-23),1.11(s,3H,H-24),1.07(s , 3H, H-26), 0.95 (s, 3H, H-25), 0.71 (s, 3H, H-28); EI (m/z): 557 (M ⁺ ), 322 (100), 542, 524, 338, 323.

Example 19: Preparation of compounds in which R4 is-CONH2 in general formula I and II:

Take 1 mmol of the compound whose R ₄ is -COOH in the general formula I or II as the raw material, dissolve it in dichloromethane (DCM), add 10 mmol of oxalyl chloride to reflux for 1 hour, concentrate under reduced pressure to remove the residual dichloromethane, then add it and dry it. 3ml of toluene was added, ammonia water was added and stirred at 4°C for reaction, and the reaction progress was detected by TLC. After the reaction is complete, water is added for direct extraction, the organic layer is dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain a crude product, which is separated and purified by silica gel column chromatography to obtain the compound in which R ₄ in the general formula I is -CONH ₂ or the compound in the general formula II Compounds where R ₄ is -CONH ₂ (such as GN-71).

Compound GN-71, yield 83.6%, light yellow powder, C ₃₀ H ₄₇ NO ₅ , m.p133～135℃.IR(KBr),σ/cm ^-1 :3433,2948,2868,1702,1646,1465 , 1385, 1217, 1116, 1046, 976, 816; ¹ H NMR (400MHz, CDCl ₃ ) δ: 5.12 (s, 1H, H-12), 4.74 (brs, 1H, H-1), 4.61 (t, 1H, J＝3.6Hz, H-2), 4.58(d, 1H, J＝4.0Hz, H-3), 2.97(s, 1H, H-9), 2.58(s, 1H, OH), 2.49(s, 1H,OH),2.49(s,1H,OH),1.28(s,3H),1.18(s,3H,H-25),1.16(s,3H,H-24),1.13(s,3H,H -27),1.12(s,3H,H-23),1.09(s,3H,H-29),0.73(s,3H,H-28); EI(m/z):501(M ⁺ ), 246(100), 483, 465, 338.

Embodiment 20: Take any one of the compounds prepared in Examples 1-19, add an appropriate amount of starch and microcrystalline cellulose according to the pharmaceutical method and mix thoroughly, mix the hydroxymethyl cellulose solution with the above-mentioned powder, and pass Wet granules obtained through 80-mesh sieve are dried at 50-60°C, carboxymethyl starch salt, magnesium stearate and talcum powder are sieved in advance, and then added to the above granules and compressed into tablets to obtain tablets. The preparation is taken orally, twice a day, and the dose of each dose is 5-30 mg based on the crude drug. For the treatment of liver damage caused by carbon tetrachloride or infectious diseases caused by Staphylococcus aureus such as purulent infection, pneumonia, pseudomembranous enteritis, pericarditis, etc.

Example 21: Take any one of the compounds prepared in Examples 1-19, add an appropriate amount of sodium citrate, polyethylene glycol, and distilled water according to the pharmaceutical method, mix and fully dissolve, and adjust the pH of the solution to 7.5-8.5 Filtrate, the concentration of the crude drug is 1mg/ml, pack in 2ml per ampoule, and sterilize to obtain the injection. The preparation is used for injection, twice a day, and the amount of each injection is 5-30 mg based on the raw drug. For the treatment of liver damage caused by carbon tetrachloride or infectious diseases caused by Staphylococcus aureus.

Embodiment 22: Get any one of the compounds prepared in Examples 1-19, pulverize through a 80 mesh sieve, add an appropriate amount of vegetable oil containing 10% beeswax according to the pharmaceutical method, and mix well; use gelatin: glycerin: distilled water: preservative =1:0.4:0.8:0.003 Prepare the capsule material, add the mixture of the aforementioned compound and vegetable oil, and press to obtain soft capsules. The preparation is taken orally, twice a day, and the dose of each dose is 5-30 mg based on the crude drug. For the treatment of liver damage caused by carbon tetrachloride or infectious diseases caused by Staphylococcus aureus.

Example 23: Take any one of the compounds prepared in Examples 1-19, crush it through an 80-mesh sieve, add an appropriate amount of polyethylene glycol according to the pharmaceutical method, mix well, and drip it into vegetable oil under heat preservation conditions to make pellets , that is, drop pills. The preparation is taken orally, twice a day, and the dose of each dose is 5-30 mg based on the crude drug. For the treatment of liver damage caused by carbon tetrachloride or infectious diseases caused by Staphylococcus aureus.

The application of the compound of formula 1 described in the present invention in pesticides is to make various dosage forms with any compound of general formula 1 plus pharmaceutically corresponding adjuvants such as surfactants, buffer reagents and adjuvants, which are applied to fungicides and antiviral agents, etc. It can be processed into formulations such as wettable powder, emulsifiable concentrate, suspension concentrate, water-dispersible granule and water-emulsion through preparation methods, and has good effects in the application of bactericide and antiviral agent.

The pesticide preparation uses any one compound with the general formula 1 as the main active ingredient, and the added surfactant can accelerate the diffusion of the active ingredient on or through the surface of leaves and stems. Cationic agents such as betaine, amido betaine, pyrrolone; anionic surfactants such as sodium lauryl sulfate; nonionic surfactants such as alkanolamide (FFA), fatty alcohol polyoxyethylene ether (AE), Alkylphenol Ethoxylates (APE or OP), Fatty Acid, Free Fatty Acid, Tween-80, Carboxymethylcellulose, Ethylene Glycol, Alkyl Ethoxylates, Alkyl Ethoxylates, Ethoxylated Soybean oil, hydrogenated castor oil, vegetable oil, silicone oil, methyl seed oil; penetration enhancers, such as pyrrole, N-alkylpyrrolidone, methylated pyrrolidone, polyvinylpyrrolidone, etc. These surfactants can be used alone or in combination.

Example 24: Weigh by weight percentage: (1) 1.00% of any one of the compounds prepared in Examples 15-19, (2) 4.00% of hydrogenated castor oil, (3) polyethylene glycol octylphenyl ether 1.00% %, (4) octylpyrrolidone 1.00%, (5) ethanol 2.00%, (6) water 91.00%; mixed to make water emulsion. When applied, the preparation is diluted 1:50-100 for the prevention and treatment of potato scab or tobacco mosaic disease.

Example 25: Weigh by weight percentage: (1) 10.00% of any compound prepared in Examples 1-19, (2) polyethylene glycol 1.00%, (3) sodium dihydrogen phosphate 2.00%, (4) Sodium dodecylsulfonate 10.00%, (5) white carbon black 5.00%, (6) starch 7.00%, (7) diatomaceous earth 65.00%; mixed to make water-dispersible granules. When applied, the preparation is diluted 1:50-100 for the prevention and treatment of potato scab or tobacco mosaic disease.

Example 26: Weighing by weight percentage: (1) 1.0% of any compound with general formula I or II, (2) 2.5% of polyethylene glycol octylphenyl ether, (3) 10.0% of tea saponin , (4) 2.0% white sugar, (5) 84.5% diatomaceous earth; mixed to make wettable powder. The preparation is diluted 1:100 when applied; it is used to control potato scab or tobacco mosaic.

Claims (16)

1. A ring polyoxygenated glycyrrhetinic acid derivatives, characterized in that: the general formula of the compound is the following structural formula 1: Among them: R ₁ is selected from carbonyl or hydroxyl; R ₂ and R ₃ are both hydroxyl or between R ₂ and R ₃ is an epoxy group; R ₄ is selected from -COOH, -COOR ¹ , -CONH ₂ , -CONHR ¹ , - any one of CONR ¹ R ² ; R ¹ and R ² are selected from one of alkyl, substituted or unsubstituted phenyl, substituted or unsubstituted phenylalkyl containing 1-15 carbon atoms, R ¹ and R ² may be the same or different; the substituents in the substituted phenyl and substituted phenylalkyl groups are selected from halogen, hydroxyl, cyano, amino, nitro, mercapto or phenyl containing 1-15 carbons, Acyl, aryl, alkoxy, alkyl; Y is CH ₂ or C=O; the 18-position hydrogen is in α configuration or β configuration; when R ₁ , R ₂ , and R ₃ are hydroxyl, the hydroxyl substitution configuration Each type is α-configuration or β-configuration; when there is an epoxy group between R ₂ and R ₃ , the epoxy group is α-configuration or β-configuration. 2. A ring polyoxidation substituted glycyrrhetinic acid derivatives according to claim 1, characterized in that: in the general formula 1, between R ₂ and R ₃ is an epoxy group, that is, the compound of formula I: 3. A ring multi-oxidation substituted glycyrrhetinic acid derivatives according to claim 2, characterized in that: in the compound of formula I, _R is selected from carbonyl or hydroxyl, and hydroxyl is α configuration or β configuration; R _and The epoxy group between R ₃ is α-configuration or β-configuration; R ₄ is selected from -COOH, benzyl ester group or benzylamino group; Y is CH ₂ or C=O; the hydrogen at position 18 is β-configuration. 4. The A-ring polyoxidized substituted glycyrrhetinic acid derivative according to claim 3, characterized in that: said compound is: Ia.3-oxo-(1α,2α)-1,2-epoxy-licorice Subacid; Ⅰb.3-Oxo-(1α,2α)-1,2-epoxy-11-deoxyglycyrrhetinic acid; Ⅰc.3-Oxo-(1α,2α)-1,2-epoxy- Benzyl glycyrrhetinate; Id.3-Oxo-(1α,2α)-1,2-epoxy-11-deoxybenzyl glycyrrhetinate; Ie.(1α,2α)-1,2-epoxy- Glycyrrhetinic acid; If.3α-Hydroxy-(1α,2α)-1,2-epoxy-11-oxo-oleanane-12-ene-30-carboxylic acid; Ig.(1α,2α)- 1,2-epoxy-11-deoxyglycyrrhetinic acid; Ⅰh.3α-hydroxy-(1α,2α)-1,2-epoxy-oleanane-12-ene-30-carboxylic acid; Ⅰi.( 1α,2α)-1,2-Epoxy-benzyl glycyrrhetinate; Ⅰj.3α-Hydroxy-(1α,2α)-1,2-epoxy-11-oxo-oleanane-12-ene -30-Benzyl carboxylate; Ⅰk.(1α,2α)-1,2-epoxy-11-deoxyglycyrrhetinic acid benzyl ester; Ⅰl.3α-hydroxy-(1α,2α)-1,2-cyclo Benzyl oxy-oleanane-12-ene-30-carboxylate; Im. 3-Oxo-(1β,2β)-1,2-epoxy-glycyrrhetinic acid; In. 3-Oxo- (1β,2β)-1,2-epoxy-11-deoxyglycyrrhetinic acid; Io.3-Oxo-(1β,2β)-1,2-epoxy-glycyrrhetinic acid benzyl ester; Ip.3- Oxo-(1β,2β)-1,2-epoxy-11-deoxyglycyrrhetinic acid benzyl ester; Ⅰq.(1β,2β)-1,2-epoxy-glycyrrhetinic acid; Ⅰr.3α-hydroxy- (1β,2β)-1,2-epoxy-11-oxo-oleanane-12-ene-30-carboxylic acid; Is.(1β,2β)-1,2-epoxy-11-deoxy Glycyrrhetinic acid; It.3α-Hydroxy-(1β,2β)-1,2-epoxy-oleanane-12-ene-30-carboxylic acid; Iu.(1β,2β)-1,2-cyclo Benzyl oxy-glycyrrhetinate; Iv. Benzyl 3α-hydroxy-(1β,2β)-1,2-epoxy-11-oxo-oleanane-12-ene-30-carboxylate; Iw .(1β,2β)-1,2-epoxy-11-deoxyglycyrrhetinic acid benzyl ester; Ⅰx.3α-hydroxy-(1β,2β)-1,2-epoxy-oleanane-12-ene -30-Benzyl carboxylate. 5. The A ring polyoxidized substituted glycyrrhetinic acid derivative according to claim 1, characterized in that: in the general formula 1, R ₁ , R ₂ , and R ₃ are all hydroxyl groups, namely the compound of formula II: 6. A ring polyoxidation substituted glycyrrhetinic acid derivatives according to claim 5, characterized in that: in the compound of formula II, the three hydroxyl groups are each in α configuration or β configuration; R ₄ is selected from -COOH or - COOR ¹ ; R ¹ is selected from one of alkyl, substituted or unsubstituted phenylalkyl containing 1-4 carbon atoms, and the substituent in the substituted phenylalkyl is selected from halogen or nitro; Y is CH ₂ or C=O; the 18-position hydrogen is in α configuration or β configuration. 7. The A-ring polyoxidized substituted glycyrrhetinic acid derivative according to claim 6, characterized in that: in the compound of formula II, R ₁ , R ₂ , and R ₃ are hydroxyl groups in α, β, and β configurations respectively, and Y is C=O; or R ₁ , R ₂ , and R ₃ are respectively α, β, and α-configured hydroxyl groups, Y is CH ₂ ; R ₄ is -COOR ¹ , and R ¹ is selected from carbon atoms containing 1-4 carbon atoms. Straight-chain alkyl or isopropyl; 18-position hydrogen is α-configuration or β-configuration. 8. The A-ring polyoxidized substituted glycyrrhetinic acid derivative according to claim 6, characterized in that: said compound is: IIa.1α, 2α, 3α-trihydroxy-11-oxo-oleanane- 12-ene-30-carboxylic acid; Ⅱb.lα,2β-dihydroxy-11-oxo-oleanane-12-ene-30-carboxylic acid; Ⅱc.lβ,2α,3α-trihydroxy-11- Oxo-oleanane-12-ene-30-carboxylic acid; Ⅱd.lβ,2β,3α-trihydroxy-11-oxo-oleanane-12-ene-30-carboxylic acid; Ⅱe.1β ,2β-dihydroxy-glycyrrhetinic acid; Ⅱf.lβ,2α-dihydroxy-glycyrrhetinic acid; Ⅱg.lα,2β,3α-trihydroxy-glycyrrhetinic acid; Ⅱh.1α,2α-dihydroxy-glycyrrhetinic acid ; Ⅱi.lα,2α,3α-trihydroxy-oleanane-12-ene-30-carboxylic acid; Ⅱj.lα,2β,3α-trihydroxy-oleanane-12-ene-30-carboxylic acid ; Ⅱk.lβ,2β,3α-trihydroxy-oleanane-12-ene-30-carboxylic acid; Ⅱl.lβ,2α,3α-trihydroxy-oleanane-12-ene-30-carboxylic acid ;Ⅱm.lβ,2β-dihydroxy-11-deoxyglycyrrhetinic acid;Ⅱn.lα,2β-dihydroxy-11-deoxyglycyrrhetinic acid;Ⅱo.lβ,2α-dihydroxy-11-deoxyglycyrrhetinic acid;Ⅱp .1α,2α-Dihydroxy-11-deoxyglycyrrhetinic acid. 9. the preparation method of A ring polyoxidation substituted glycyrrhetinic acid derivative as claimed in claim 2 is characterized in that: glycyrrhetinic acid is dissolved with DMF, i.e. N,N-dimethylformamide, and adds potassium carbonate powder , the benzyl bromide and the carboxyl group are formed into esters to obtain the benzyl compound G-1; G-1 is dissolved in absolute ethanol, zinc powder is added and hydrochloric acid is added dropwise, and the 11-position carbonyl is reduced by zinc powder to obtain Gb; G-1, Gb Dissolved in acetone respectively, and oxidized by Jones reagent to obtain compounds G-2 and Gc; G-2 and Gc were respectively dissolved in acetic anhydride and oxidized by selenium dioxide to obtain α, β-unsaturated ketone compounds G-3 and Gd; G -3. Gd is dissolved in methanol, added with 10% NaOH solution, oxidized by H ₂ O ₂ into epoxy compounds Ic, Id and isomers Io, Ip; Ii, Ij, Ik, Il, Iu, Iv, Iw, Ix were obtained by reduction; benzyl ester compounds Ic, Id, Io, Ip, Ii, Ij, Ik, Il, Iu, Iv, Iw, Ix were dissolved in methanol respectively, and the Catalytic hydrogenation of palladium carbon and hydrogen to remove benzyl groups to obtain compounds Ia, Ib, Im, In, Ie, If, Ig, Ih, Iq, Ir, Is, It in the formula I where R is _-COOH ; the reaction scheme is as follows: 10. The preparation method of A ring multi-oxidation substituted glycyrrhetinic acid derivatives as claimed in claim 2, characterized in that: the compound in which R in the formula I _is -COOH is used as raw material, and DMF, i.e. N,N-di Methylformamide is dissolved and reacted with the corresponding haloalkane R ¹ X under the action of potassium carbonate to obtain the corresponding ester compound, that is, the compound in which R ₄ is -COOR ¹ in formula I; the raw material is N,N-dimethylformamide Dissolve and react with the corresponding amine R ¹ NH ₂ , R ¹ R ² NH under the action of DCC, namely N, N'-dicyclohexylcarbodiimide and DMAP, namely 4-dimethylaminopyridine, to obtain the corresponding amide Compounds, that is, compounds in which R ₄ is -CONHR ¹ , -CONR ¹ R ² in formula I; the raw material is dissolved in DCM, that is, dichloromethane, reacted with oxalyl chloride to form an acid chloride compound, and then the acid chloride compound is dissolved in toluene, and ammonia water is added Reaction in the formula I R ₄ is-CONH ₂ compounds; its reaction scheme is as follows: 11. The preparation method of A-ring polyoxidized substituted glycyrrhetinic acid derivatives as claimed in claim 5, characterized in that: the compounds Ii, Ij, Iu, Iv, Ik, Il, Iw, Ix are respectively used as raw materials, dissolved in Add distilled water to acetone, open the ring with perchloric acid to obtain the corresponding benzyl-substituted ring-opened compound, then dissolve the ring-opened compound in methanol, and undergo catalytic hydrogenation with palladium carbon and hydrogen to obtain the compound in which _R in the formula II is -COOH; The reaction scheme is as follows: 12. The preparation method of A-ring polyoxidatively substituted glycyrrhetinic acid derivatives as claimed in claim 5, characterized in that: the compounds IIa～IIp are used as raw materials, dissolved in N,N-dimethylformamide, and dissolved in potassium carbonate Under the influence of the reaction with the corresponding haloalkane R ¹ X to obtain the corresponding ester compound, that is, the compound in which R ₄ is -COOR ¹ in formula II; the raw material is dissolved in N,N-dimethylformamide, and the N,N'-di Cyclohexylcarbodiimide and 4-dimethylaminopyridine react with the corresponding amines R ¹ NH ₂ , R ¹ R ² NH to obtain the corresponding amide compounds, that is, R ₄ in formula II is -CONHR ¹ , - The compound of CONR ¹ R ² ; the raw material is dissolved in dichloromethane, reacted with oxalyl chloride to generate an acid chloride compound, and then the acid chloride compound is dissolved in toluene, and ammonia water is added to react to obtain a compound in formula II where R ₄ is -CONH ₂ ; the reaction scheme is as follows : 13. The use of the A-ring polyoxygenated glycyrrhetinic acid derivatives and their pharmaceutically acceptable salts according to any one of claims 1-8 in the preparation of medical drugs for treating liver damage diseases. 14. The application of the A-ring polyoxidized substituted glycyrrhetinic acid derivatives and their pharmaceutically acceptable salts in the preparation of antibacterial or anti-tobacco mosaic virus drugs according to any one of claims 1-8. 15. The application according to claim 14, characterized in that: the antibacterial drug is an antibacterial or antifungal drug; the anti-tobacco mosaic virus drug is used to prevent and treat tobacco diseases caused by tobacco mosaic virus agricultural drugs. 16. The application according to claim 15, characterized in that: the antibacterial drug is an agricultural drug for preventing and treating crop diseases caused by Gram-positive bacteria or treating infectious diseases caused by Gram-positive bacteria The medical drug; the antifungal drug is an agricultural drug for preventing and treating crop diseases caused by tobacco black shank bacteria.