Application of lindley eupatorium herb in preparing anti-hepatitis B virus medicaments
An anti-hepatitis B, wild horse technology, used in antiviral agents, drug combinations, pharmaceutical formulations, etc.
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Embodiment 1
[0030] Example 1: Preparation of Four Groups of Yemazhui Extracts and Chain Diterpene Compounds in Yemazhui
[0031] (1) Weigh 1000g of Yemazhui dried whole herb, cut into sections of 2-3cm, add 15 times the volume of 80% ethanol aqueous solution, reflux extraction for 2 hours, repeat twice, recover the solvent under reduced pressure at 60°C to obtain Yemazhui Ethanol extract (EUP-EtOH) 158g, dissolved in about 150ml water, adsorbed by 1800ml AB-8 macroporous adsorption resin, first eluted with 20000ml water to remove macromolecular compounds, and then eluted with 20000ml volume fraction 95% ethanol aqueous solution, The solvent was recovered under reduced pressure at 60°C to obtain 61g of macroporous resin parts; the obtained macroporous resin parts were adsorbed by 1500ml Cosmsil ODS reversed-phase silica gel filler particle size 70um reversed-phase silica gel chromatography column, and eluted with 10000ml volume fraction as 50% methanol aqueous solution, 60 The solvent was ...
Embodiment 2
[0033] Example 2: Detection of active ingredients of sesquiterpene parts, flavonoid parts and chain diterpene compounds in Yemazhui
[0034] (1) Detection of the active ingredient in the sesquiterpene part of Yemazhui (EUP-SQT): In this example, high performance liquid chromatography was used to determine its ingredients. High performance liquid chromatography (Agilent1260Infinity, Agilent Corporation of the United States), wherein mobile phase A is acetonitrile; mobile phase B is water; mobile phase conditions are shown in Table 1. Chromatographic column: YMC-PACK ODS-A (Ф4.6mm×250mm, 5μm); column temperature: 28℃; detection wavelength: 210nm; injection volume: 20μl; flow rate: 1ml min -1 .
[0035] Table 1 HPLC chromatographic analysis conditions
[0036] time (min)
Mobile phase A(%)
Mobile phase B(%)
0
30
70
5
30
70
15
32
68
25
36
64
40
36
64
[0037] Accurately weigh 6.02 ...
Embodiment 3
[0063]Example 3: Anti-hepatitis B virus HBV experiment of four kinds of Yemazhui extracts and chain diterpene compounds in Yemazhui
[0064] 1. Experimental drugs and reagents: chain diterpene compounds (1) and (2) prepared in the above-mentioned embodiment 1; dimethyl sulfoxide; MTT (thiazolium blue), Shanghai Jianglai Biotechnology Co., Ltd.; reference drug, Lamy Furidin Tablets (3TC); GlaxoSmithKline Pharmaceuticals (Suzhou) Co., Ltd.; cell culture medium DMEM, growth fluid, prepared, containing 10% fetal bovine serum, 380 μg / ml G418, 0.03% glutamine, penicillin, streptamine Mycin each 100μg / ml, etc.;
[0065] 2. Experimental equipment: ELISA kits for detecting hepatitis B surface antigen HBsAg and hepatitis B core antigen HBeAg; incubators; 24-well plates, etc.
[0066] 3. Cell line: 2.2.15 cell line of Hep G2, Ministry of Education / Ministry of Health Key Laboratory of Medical Molecular Virology, Shanghai.
[0067] 4. Experimental method:
[0068] Take the 2.2.15 cell l...
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