Matrine type alkaloid SA as well as preparation method and application thereof

Abstract

The invention discloses matrine type alkaloid SA as well as a preparation method and application thereof, and belongs to the field of traditional Chinese medicine preparation. The matrine type alkaloid with a new structure is extracted from sophora moorcroftiana, and cell experiments find that the matrine type alkaloid not only has the effect of resisting hepatitis B virus (HBV), but also has the inhibitory effect on hepatitis B surface antigen (HBsAg) superior to that of matrine and oxymatrine of the same kind, and also superior to that of lamivudine. Therefore, the matrine type alkaloid can be used for preparing the medicine for treating or preventing hepatitis B virus (HBV) infection.

Description

Technical Field

[0001] The invention relates to a matrine-type alkaloid SA and a preparation method and application thereof, belonging to the field of traditional Chinese medicine pharmacy. Background Art

[0002] Infection with hepatitis B virus (HBV) can cause acute and chronic hepatitis B, and is one of the important factors leading to liver cirrhosis and hepatocellular carcinoma. According to statistics, 2 billion people have been infected with HBV in the world. Every year, there are about 1 million deaths due to liver cirrhosis, liver failure and primary hepatocellular carcinoma (HCC) caused by HBV infection. Hepatitis B has become one of the top ten causes of death in the world. HBV infection can develop into chronic hepatitis, cirrhosis, liver failure and liver cancer, which seriously threatens human life and health. At present, clinical anti-HBV drugs are mainly interferon and nucleoside drugs (lamivudine, adefovir dipivoxil, entecavir, telbivudine, tenofovir, etc.). Although these two types of drugs have achieved relatively good therapeutic effects in the treatment of HBV infection, it has been reported that interferon has poor tolerance, many side effects, and a narrow range of indications. It can only relieve the condition of 20-50% of chronic hepatitis B patients for a long time, and its side effects also limit its application. Although nucleoside drugs have a strong anti-HBV effect, they are prone to relapse after discontinuation of the drug, and long-term use can easily lead to drug resistance. Inappropriate discontinuation of the drug can lead to recurrence or deterioration of the disease, and some patients are prone to adverse reactions such as virus mutation, nephrotoxicity, and bone marrow suppression, which leads to their inability to be widely used in clinical practice. Currently, there is no stable and ideal anti-HBV drug in clinical practice. Therefore, the development of new anti-HBV drugs has always been the focus of effective prevention and treatment of hepatitis B. Summary of the invention

[0003] In order to more effectively treat hepatitis B virus, one of the purposes of the present invention is to provide a matrine-type alkaloid SA, the structure of which is shown in Formula I:

[0004]

[0005] Preferably, the matrine-type alkaloid SA is obtained from Sophora morabica seeds by extraction, separation or artificial synthesis.

[0006] The second object of the present invention is to provide a method for preparing matrine-type alkaloid SA, comprising the following steps:

[0007] (1) Sophora moorii seeds are dried, crushed, and then cold-extracted with a mixed solution of ethanol and water. The extracts are combined and concentrated by vacuum distillation to obtain an extract.

[0008] (2) The extract is distributed in a hydrochloric acid aqueous solution, and then extracted with ethyl acetate. The pH value of the water-soluble part after extraction is adjusted with an ammonia aqueous solution, and then extracted with chloroform to obtain a total alkaloid extract.

[0009] (3) Matrine-type alkaloid SA was isolated from the alkaloid extract.

[0010] Preferably, in step (1), the volume ratio of ethanol to water is 95:5, and the number of cold soaking extractions is 3 times.

[0011] Preferably, in step (2), the pH of the hydrochloric acid aqueous solution is 1-2, extracted with ethyl acetate three times, the pH value is adjusted to 8-9 with an ammonia aqueous solution, and extracted with chloroform three times.

[0012] Preferably, the separation in step (3) comprises the following steps:

[0013] (a) The alkaloid extract was separated by normal silica gel, and petroleum ether-ethyl acetate was first used as the mobile phase with a volume ratio of 9:1 to 1:1 to obtain F0, F1, F2, and F3, and then petroleum ether-acetone was used as the mobile phase with a volume ratio of 5:1 to 1:1 to obtain F4, F5, and F6. Finally, chloroform-methanol was used as the mobile phase with a volume ratio of 9:1 to 1:1 to obtain F7, F8, F9, F10, and F11. All samples were rotated, concentrated, and weighed in turn to obtain 12 polar segments F0, F1, F2, F3, F4, F5, F6, F7, F8, F9, F10, and F11.

[0014] (b) The F11 segment obtained by eluting with chloroform-methanol volume ratio of 3:1 in step (a) was passed through a normal silica gel column with petroleum ether-acetone-diethylamine in a volume ratio of 100:10:1 to 20:10:1 as the mobile phase to obtain 7 segments, namely F11-1, F11-2, F11-3, F11-4, F11-5, F11-6 and F11-7.

[0015] (c) The component F11-2 obtained by eluting with petroleum ether-acetone-diethylamine in a volume ratio of 100:10:1 in step (b) was passed through a normal silica gel column and eluted with chloroform-methanol in a volume ratio of 10:1 to 1:1 as the mobile phase to obtain four parts, namely F11-2-1, F11-2-2, F11-2-3 and F11-2-4.

[0016] (d) The F11-2-1 segment obtained by eluting with chloroform-methanol volume ratio of 10:1 in step (c) was separated by a gel column with methanol as the elution condition, and then passed through a forward silica gel column with chloroform-acetone as the mobile phase in a volume ratio of 100:1 to obtain matrine-type alkaloid SA.

[0017] Preferably, in step (a), F0 is eluted with a petroleum ether-ethyl acetate volume ratio of 9:1, F1 is eluted with a petroleum ether-ethyl acetate volume ratio of 5:1, F2 is eluted with a petroleum ether-ethyl acetate volume ratio of 1:1, F3 is eluted with a petroleum ether-ethyl acetate volume ratio of 1:1, F4 is eluted with a petroleum ether-acetone volume ratio of 5:1, F5 is eluted with a petroleum ether-acetone volume ratio of 3:1, F6 is eluted with a petroleum ether-acetone volume ratio of 1:1, F7 is eluted with a chloroform-methanol volume ratio of 9:1, F8 is eluted with a chloroform-methanol volume ratio of 9:1, F9 is eluted with a chloroform-methanol volume ratio of 6:1, F10 is eluted with a chloroform-methanol volume ratio of 3:1, and F11 is eluted with a chloroform-methanol volume ratio of 3:1.

[0018] Preferably, in step (b), F11-1 is eluted by petroleum ether-acetone-diethylamine in a volume ratio of 100:10:1, F11-2 is eluted by petroleum ether-acetone-diethylamine in a volume ratio of 100:10:1, F11-3 is eluted by petroleum ether-acetone-diethylamine in a volume ratio of 80:10:1, F11-4 is eluted by petroleum ether-acetone-diethylamine in a volume ratio of 60:10:1, F11-5 is eluted by petroleum ether-acetone-diethylamine in a volume ratio of 60:10:1, F11-6 is eluted by petroleum ether-acetone-diethylamine in a volume ratio of 40:10:1, and F11-7 is eluted by petroleum ether-acetone-diethylamine in a volume ratio of 20:10:1.

[0019] Preferably, in step (c), F11-2-1 is obtained by eluting with a chloroform-methanol volume ratio of 10:1, F11-2-2 is obtained by eluting with a chloroform-methanol volume ratio of 8:1, F11-2-3 is obtained by eluting with a chloroform-methanol volume ratio of 5:1, and F11-2-4 is obtained by eluting with a chloroform-methanol volume ratio of 3:1.

[0020] The third object of the present invention is to provide a medicine.

[0021] Preferably, the drug comprises matrine-type alkaloid SA.

[0022] Preferably, the drug is a microparticle drug delivery system, and the dosage form is tablets, capsules, pills, injections, sustained-release preparations, and controlled-release preparations.

[0023] Preferably, the drug is obtained by combining matrine-type alkaloid SA and at least one pharmaceutically acceptable carrier.

[0024] Preferably, the combination drug is a drug consisting of matrine-type alkaloid SA and a pharmaceutically acceptable salt.

[0025] Preferably, the pharmaceutically acceptable salts include: citric acid, maleic acid, fumaric acid, hydrochloric acid, sulfuric acid, phosphoric acid, lithium, sodium, potassium, calcium, magnesium or lysine.

[0026] The fourth object of the present invention is to provide a use of the matrine-type alkaloid SA or drug in the preparation of a drug for treating or preventing hepatitis B virus infection.

[0027] Beneficial effects of the present invention

[0028] (1) The present invention provides a new matrine-type alkaloid SA, wherein the matrine-type alkaloid SA has C 15 The alkaloids have an N2 carbon skeleton, and their structural characteristics are that the A, B and C rings form an unusual decahydro-1H-pyrrolo[3,2,1-ij][1,6]naphthyridine structural unit, and the B, C and D rings form a dodecahydropyrrolo[3,2-a]quinolinazine structure that is even rarer in natural products.

[0029] (2) The matrine-type alkaloid SA provided by the present invention can not only be used to prepare drugs for treating or preventing hepatitis B, but also has better therapeutic effects than existing matrine and oxymatrine. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is a schematic diagram of the structure of the matrine-type alkaloid SA of the present invention.

[0031] Figure 2 It is a schematic diagram of the single crystal X-ray diffraction structure of the matrine-type alkaloid SA of the present invention.

[0032] Figure 3 The present invention is a flow chart of the preparation method of the matrine-type alkaloid SA. DETAILED DESCRIPTION

[0033] The technical solution of the present invention is further described below in conjunction with the accompanying drawings and through specific implementation methods. However, the following embodiments are only simple examples of the present invention and do not represent or limit the scope of protection of the present invention. The scope of protection of the present invention shall be subject to the claims.

[0034] Example 1

[0035] A preparation method and structural identification of a matrine-type alkaloid SA, the specific steps are as follows:

[0036] (1) Separation process: Take the dried seeds of Tibetan medicine Sophora japonica (20 kg), grind them, and use a mixed solution of ethanol and water with a volume ratio of 95:5 to cold-extract them three times at room temperature. Combine the extracts, and concentrate them under reduced pressure to remove the organic solvent to obtain an extract.

[0037] (2) The extract was distributed in a hydrochloric acid aqueous solution at a pH of 2, and then extracted three times with ethyl acetate to remove most of the non-alkaloid components; the water-soluble part after extraction was adjusted to pH 8.2 with an ammonia solution, and then fully extracted three times with chloroform to obtain a total alkaloid extract (826.7 g).

[0038] (3) The alkaloid extract was separated by normal silica gel, and petroleum ether-ethyl acetate was first used as the mobile phase with a volume ratio of 9:1, 5:1, 1:1, and 1:1 to obtain F0, F1, F2, and F3; petroleum ether-acetone was then used as the mobile phase with a volume ratio of 5:1, 3:1, and 1:1 to obtain F4, F5, and F6; finally, chloroform-methanol was used as the mobile phase with a volume ratio of 9:1, 9:1, 6:1, 3:1, and 3:1 to obtain F7, F8, F9, F10, and F11.

[0039] (4) The F11 segment (54.6 g) was passed through a normal silica gel column and eluted with petroleum ether-acetone-diethylamine as the mobile phase at a volume ratio of 100:10:1, 100:10:1, 80:10:1, 60:10:1, 40:10:1, and 20:10:1 to obtain 7 segments, namely F11-1, F11-2, F11-3, F11-4, F11-5, F11-6, and F11-7.

[0040] (5) F11-2 fragment (345.4 mg) was passed through a normal silica gel column and eluted with chloroform-methanol mobile phase at volume ratios of 10:1, 8:1, 5:1, and 3:1 to obtain four fractions, namely F11-2-1, F11-2-2, F11-2-3, and F11-2-4.

[0041] (6) The F11-2-1 (33.9 mg) fragment was separated by a gel column with methanol as the elution condition to remove impurities, and then passed through a normal silica gel column with chloroform-acetone volume ratio of 100:1 as the mobile phase to obtain a compound SA (5.0 mg).

[0042] Through the analysis of MS and NMR spectral data (Table 1), the molecular formula of compound SA is C 14 H 18 N2O2 is a colorless needle crystal with a melting point of 108-109°C. Its molecular ion peak in high-resolution mass spectrometry (HR-ESI-MS) is m / z 249.1596 [M+H] + (calculated value is 249.1598), the structure of compound SA is as follows Figure 1 As shown, it is a matrine-type alkaloid.

[0043] Finally, the stereo configuration of the matrine-type alkaloid SA (Sophormodine A) was further determined by X-ray single crystal diffraction analysis. Figure 2 shown.

[0044] Table 1 NMR data of matrine-type alkaloids SA ( 1 H and 13 C NMR spectra were recorded at600 and 150MHz, respectively.in methanol-d4)

[0045] serial number <![CDATA[δ H (J in Hz)]]> <![CDATA[δ C ]]> serial number <![CDATA[δ H (J in Hz)]]> <![CDATA[δ C ]]> 2a 3.92(d，13.2) 41.4 9 - 171.8 2b 2.62(d，13.2) - 11 3.12(dt,10.8,5.3) 57.1 3a 2.02(m) 27.1 12a 1.76(m) 26.8 3b 1.64(m) - 12b 1.76(m) - 4a 1.54(m) 19.7 13a 1.80(m) 18.7 4b 1.54(m) - 13b 1.65(m) - 5 1.94(m) 33.0 14a 2.28(m) 33.2 6 3.67(t，4.9) 58.8 14b 2.28(m) - 7 2.33(m) 39.0 15 - 177.0 8a 2.47(m) 36.5 17α 4.26 (dd, 13.0, 4.9) 42.4 8b 2.00(m) - 17β 2.69(d，13.0) -

[0046] Example 2

[0047] Example 1 Verification of the anti-HBV activity of matrine-type alkaloid SA prepared

[0048] 1. HepG 2.2.15 cell culture

[0049] (1) Cell screening model: HepG2.2.15 cells were purchased from Guangzhou Saiku Biotechnology, subcultured by our laboratory, and stored in liquid nitrogen and a -80°C freezer.

[0050] (2) Culture medium: Add 10% non-essential amino acids and 380 μg / mL G418 (aminoglycoside antibiotic) to 500 mL of MEM culture medium. Prepare before the experiment and store in a refrigerator at 4°C until use.

[0051] (3) Cell recovery: Before the experiment, prepare complete culture medium in a clean bench. The complete culture medium includes 90% culture medium + 10% FBS. Shake it up and down to mix it thoroughly, and heat it in a 37°C water bath. Take 5 mL of the complete culture medium after warming up and put it in a T25 culture bottle for use. At this time, take out a frozen HepG2.2.15 cell from the liquid nitrogen tank, quickly put it in a 37°C water bath, and continue to shake it gently. After it is completely thawed, transfer the cell suspension to a 15 mL centrifuge tube, and add 8 mL of culture medium that has been restored to room temperature dropwise, while mixing. Centrifuge at 8000 rpm for 3 minutes at room temperature, discard the supernatant, add enough culture medium to resuspend, and inoculate the cell suspension into a T25 culture bottle containing 5 mL of complete culture medium. Replace with fresh culture medium after 24 hours, and then change the medium every 3 days. After about 5 days, the cells will adhere to the wall and grow full.

[0052] (4) Cell passaging: When the cell growth density reaches 90%, remove the cells from the incubator, discard the original culture medium, add 2 mL of preheated PBS each time, wash three times, then add 1 mL of trypsin, gently shake to allow all cells to fully contact, aspirate the trypsin, put the cells into the incubator for digestion for 3 minutes, and observe under a microscope. Obvious changes in cell morphology were observed. At this time, 2 mL of complete culture medium was added to terminate digestion, the cells were blown off, transferred to a 5 mL eppendorf tube, centrifuged at 8000 rpm for 3 minutes, the supernatant was discarded, 1 mL of culture medium was added, and the mixture was blown evenly. 330 μL of single-cell suspension was inoculated into each dish, 5 mL of culture medium was added, and the mixture was blown evenly and put into the incubator.

[0053] (5) Cell freezing: Prepare freezing solution, 10% DMSO + 90% complete medium (prepared and used immediately), where the total volume percentage of DMSO and complete medium is 100. Select cells (F3 / F4) with good growth status, remove them from the incubator, discard the original medium, add 2mL of PBS, wash 3 times, then add 1mL of trypsin to digest the cells, put them in the incubator for digestion for 3min, and then check under a microscope, add 2mL of complete medium to stop digestion, blow down the cells, transfer them to an eppendorf tube, centrifuge them at 8000rpm for 3min, discard the supernatant, add 1mL of the prepared freezing solution, and indicate the time, cell generation, freezing solution, cell name and operator in the freezing tube. Perform gradient cooling freezing: place at 4℃ for 30min→-20℃ for 2h→-80℃ overnight, and place them in liquid nitrogen the next day.

[0054] 2. MTT assay for cytotoxicity

[0055] Cell plate: Select HepG2.2.15 cells in good growth stage, with a cell density of 1×10 5 Cells were inoculated into 24-well plates, 500 μl per well, and placed in an incubator for culture. After 24 hours, drugs were administered (so that the final concentration of matrine-type alkaloids SA, matrine and oxymatrine in the cell culture wells was 0.6 mmol / L, and the final concentration of 3TC (lamivudine) in the cell culture wells was 1 mmol / L), of which lamivudine was positive. A blank control group without drug administration was set up at the same time. The supernatant was collected and replaced with fresh drug solution every 3 days, and the drug was added and cultured for a total of 6 days. At 6 days, the supernatant was aspirated and placed in a -80℃ refrigerator for standby use. After adding 200 μL of PBS to each well, 50 μl of MTT was added, and after being placed in an incubator for 4 hours, the supernatant was discarded, and 500 μL of DMSO was added to each well to measure its absorbance at 490 nm.

[0056] Cell survival rate = A 490 (Sample) / A 490 (Control)×100%

[0057] Among them A490 (Sample) is the absorbance value of adding matrine alkaloid SA, matrine, oxymatrine or positive drug, A 490 (Control) is the absorbance value of blank control. IBM SPSS Statistics 20 was used to analyze the experimental data.

[0058] 3. ELISA method to detect HBsAg content in cell supernatant

[0059] HBsAg test kit - Shanghai Kehua Bioengineering Co., Ltd., product batch number 202310271. All operations were performed according to the instructions. The specific steps are as follows:

[0060] (1) Take the test kit out of the 4°C refrigerator and equilibrate it to room temperature;

[0061] (2) Working concentration washing solution: dilute the washing solution (25×) in the kit to the working concentration. For example, to prepare 500 mL of working solution, add 480 mL of ultrapure water + 20 mL of washing solution (25×).

[0062] Operation steps: according to the number of samples, select a certain amount of reaction strips equilibrated to room temperature, add 75 μL of supernatant (the supernatant here is the supernatant saved in the MTT method cytotoxicity test step (2)) to the reaction well, set up three duplicate wells: 3 negative control wells (normal human serum), 1 positive control well and 1 blank control well, cover the reaction plate with sealing paper, and incubate at 37°C for 1 hour; then take out the reaction plate, add 50 μL of enzyme conjugate (anti-Hbs antibody labeled with horseradish peroxidase) to each well, shake gently, cover with sealing paper, and incubate at 37°C for 30 minutes; take out the reaction plate, add the prepared working washing solution Add the solution into the wells, fill it up (add slowly and gently), let it stand for 1 min, discard the liquid in the wells (you can use a pipette to suck it out, but try not to touch the bottom of the wells), pat dry on absorbent paper after each spin drying (the amplitude and force of the movement should not be too large when patting dry), repeat 5 times; mix the color developer A and color developer B in a volume ratio of 1:1 for standby use, add 100 μL of the prepared color developer into the wells immediately after washing, shake gently, cover with sealing paper, incubate at 37°C for 30 min, add 50 μL of stop solution to each well after the reaction is completed to terminate the reaction; use an enzyme marker to detect at wavelengths of 450nm and 630nm, and determine the OD value of each well.

[0063] HBsAg inhibition rate = [(A 450 Positive control group-A 450 Medication group) / A 450 Positive control group] × 100%

[0064] Test validity: If NCx≤0.100 and PC≥1.000, the test result is valid.

[0065] NCx: Negative control average OD value

[0066] PC: Negative control OD value

[0067] IBM SPSS Statistics 20 was used to analyze the experimental data. The experimental data were repeated three times and the results were expressed as mean ± standard deviation (SD). The final calculation results are shown in Table 2.

[0068] 4. ELISA method to detect HBeAg content in cell supernatant

[0069] HBeAg test kit - Shanghai Kehua Bioengineering Co., Ltd.; product batch number 202310172. All operations were performed according to the instructions. The specific steps are as follows:

[0070] According to the number of samples, select a certain amount of reaction strips equilibrated to room temperature, add 50 μL of supernatant (the supernatant here is the supernatant saved in the MTT method cytotoxicity test step (2)) to the reaction wells, set up three duplicate wells: 2 negative control wells (normal human serum), 2 positive control wells and 1 blank control well; add 50 μL of enzyme conjugate (anti-Hbe antibody labeled with horseradish peroxidase) to each well, shake gently, cover with sealing paper, and incubate at 37°C for 30 minutes; add the prepared working washing solution to the wells, fill them up (add the solution gently and slowly), let them stand for 5 seconds, and discard the liquid in the wells (can be removed). Aspirate with a pipette, but try not to touch the bottom of the well), pat dry on absorbent paper after each spin (the amplitude and force of the movement should not be too large when patting dry), repeat 5 times; mix the color developer A and the color developer B in a volume ratio of 1:1 to obtain the color developer for standby use (the color developer A and the color developer B are the reagents in the kit), add 100 μL of the prepared color developer to the well immediately after the washing, shake gently, cover with sealing paper, incubate at 37°C for 15 minutes, and add 50 μL of stop solution to each well after the reaction is completed to terminate the reaction; use an enzyme marker to detect at wavelengths of 450nm and 630nm, and determine the OD value of each well.

[0071] HBeAg inhibition rate = [(A 450 Positive control group-A 450 Medication group) / A 450 Positive control group] × 100%

[0072] Reference value cut-off value (COV) = negative control average OD value × 2.1

[0073] Negative OD values ​​below 0.050 are calculated as 0.050, and those 0.050≤OD≤0.100 are calculated according to the actual OD value.

[0074] When the negative control OD value is greater than 0.100, the result is invalid and the experiment needs to be repeated.

[0075] When the OD value of the sample is less than the COV value, it means that the HBeAg test result of the sample is negative.

[0076] When the OD value of the sample is ≥COV value, it means that the HBeAg test result of the sample is positive.

[0077] IBM SPSS Statistics 20 was used to analyze the experimental data. The experimental data were repeated three times and the results were expressed as mean ± standard deviation (SD). The final calculation results are shown in Table 2.

[0078] Results Analysis

[0079] The results of HepG2.2.15 cell experiments showed that the matrine-type alkaloid SA has a good anti-HBV effect in vitro, especially the inhibitory effect on hepatitis B surface antigen (HBsAg) is better than matrine and oxymatrine, and is also better than the positive control drug lamivudine (3TC).

[0080] Table 2 Screening results of anti-HBV activity of matrine-type alkaloid SA

[0081]

[0082] (1) The cytotoxic effects of 0.6 mmol / L of matrine-type alkaloids SA, matrine, oxymatrine and 1.0 mmol / L of 3TC were determined by MTT assay. The cell survival rates were all greater than 85%, indicating no cytotoxic effects.

[0083] (2) The above experiments prove that the matrine-type alkaloids prepared by the present invention can significantly resist hepatitis B virus and have little toxicity to cells. Therefore, they can be used to prepare drugs for preventing or resisting hepatitis B virus.

Claims

1. A matrine-type alkaloid SA, characterized in that: The structure of the matrine-type alkaloid SA is shown in Formula I:

2. The matrine-type alkaloid SA according to claim 1, characterized in that: The matrine-type alkaloid SA is obtained from Sophora morafum seeds by extraction, separation or artificial synthesis.

3. The method for preparing the matrine-type alkaloid SA according to claim 1 or 2, characterized in that: The following steps are involved: (1) drying and crushing the seeds of Sophora moracha, cold-immersing and extracting them with a mixed solution of ethanol and water, combining the extracts, and concentrating the extracts by vacuum distillation to obtain an extract; (2) distributing the extract in a hydrochloric acid aqueous solution, then extracting with ethyl acetate, adjusting the pH value of the water-soluble part after extraction with an ammonia aqueous solution, and then extracting with chloroform to obtain a total alkaloid extract; (3) Matrine-type alkaloid SA was isolated from the alkaloid extract.

4. The method for preparing the matrine-type alkaloid SA according to claim 3, characterized in that: In step (1), the volume ratio of ethanol to water is 95:5, and the number of cold soaking extractions is 3 times.

5. The method for preparing matrine-type alkaloid SA according to claim 3, characterized in that: In step (2), the pH of the hydrochloric acid aqueous solution is 1-2, and the solution is extracted with ethyl acetate three times, the pH value is adjusted to 8-9 with an ammonia aqueous solution, and the solution is extracted with chloroform three times.

6. The method for preparing matrine-type alkaloid SA according to claim 3, characterized in that: The separation in step (3) comprises the following steps: (a) The alkaloid extract was separated by normal silica gel, and petroleum ether-ethyl acetate was first used as the mobile phase with a volume ratio of 9:1 to 1:1 to obtain F0, F1, F2, and F3, and then petroleum ether-acetone was used as the mobile phase with a volume ratio of 5:1 to 1:1 to obtain F4, F5, and F6, and finally chloroform-methanol was used as the mobile phase with a volume ratio of 9:1 to 1:1 to obtain F7, F8, F9, F10, and F11. All samples were rotated, concentrated, and weighed in turn to obtain 12 polar segments F0, F1, F2, F3, F4, F5, F6, F7, F8, F9, F10, and F11; (b) passing the F11 segment obtained by eluting with chloroform-methanol volume ratio of 3:1 in step (a) through a normal silica gel column, and eluting with petroleum ether-acetone-diethylamine volume ratio of 100:10:1 to 20:10:1 as the mobile phase to obtain 7 segments F11-1, F11-2, F11-3, F11-4, F11-5, F11-6 and F11-7; (c) The component F11-2 obtained by eluting with petroleum ether-acetone-diethylamine in a volume ratio of 100:10:1 in step (b) was passed through a normal silica gel column, and the elution condition was chloroform-methanol in a volume ratio of 10:1 to 1:1 as the mobile phase to obtain 4 parts F11-2-1, F11-2-2, F11-2-3 and F11-2-4; (d) The F11-2-1 segment obtained by eluting with chloroform-methanol volume ratio of 10:1 in step (c) was separated by a gel column with methanol as the elution condition, and then passed through a forward silica gel column with chloroform-acetone as the mobile phase in a volume ratio of 100:1 to obtain matrine-type alkaloid SA.

7. The method for preparing matrine-type alkaloid SA according to claim 6, characterized in that: In step (a), F0 is eluted by petroleum ether-ethyl acetate volume ratio of 9:1, F1 is eluted by petroleum ether-ethyl acetate volume ratio of 5:1, F2 is eluted by petroleum ether-ethyl acetate volume ratio of 1:1, F3 is eluted by petroleum ether-ethyl acetate volume ratio of 1:1, F4 is eluted by petroleum ether-acetone volume ratio of 5:1, F5 is eluted by petroleum ether-acetone volume ratio of 3:1, F6 is eluted by petroleum ether-acetone volume ratio of 1:1, F7 is eluted by chloroform-methanol volume ratio of 9:1, F8 is eluted by chloroform-methanol volume ratio of 9:1, F9 is eluted by chloroform-methanol volume ratio of 6:1, F10 is eluted by chloroform-methanol volume ratio of 3:1, and F11 is eluted by chloroform-methanol volume ratio of 3:

1.

8. The method for preparing matrine-type alkaloid SA according to claim 6, characterized in that: In step (b), F11-1 is eluted by a petroleum ether-acetone-diethylamine volume ratio of 100:10:1, F11-2 is eluted by a petroleum ether-acetone-diethylamine volume ratio of 100:10:1, F11-3 is eluted by a petroleum ether-acetone-diethylamine volume ratio of 80:10:1, F11-4 is eluted by a petroleum ether-acetone-diethylamine volume ratio of 60:10:1, F11-5 is eluted by a petroleum ether-acetone-diethylamine volume ratio of 60:10:1, F11-6 is eluted by a petroleum ether-acetone-diethylamine volume ratio of 40:10:1, and F11-7 is eluted by a petroleum ether-acetone-diethylamine volume ratio of 20:10:

1.

9. The method for preparing matrine-type alkaloid SA according to claim 6, characterized in that: In step (c), F11-2-1 is obtained by eluting with a chloroform-methanol volume ratio of 10:1, F11-2-2 is obtained by eluting with a chloroform-methanol volume ratio of 8:1, F11-2-3 is obtained by eluting with a chloroform-methanol volume ratio of 5:1, and F11-2-4 is obtained by eluting with a chloroform-methanol volume ratio of 3:

1.

10. Use of the matrine-type alkaloid SA according to claim 1 or the drug according to claim 9 in the preparation of a drug for treating or preventing hepatitis B virus infection.

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