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Method of preparing extract of Mallotus apelta and uses for fighting hepatitis B virus

The technology of hepatitis B virus and extract is applied in the field of preparation of white back leaf extract and the application of anti-hepatitis B virus, and achieves the effects of significant anti-hepatitis B virus effect, inhibition of DNA replication of hepatitis B virus, and simple and easy preparation method.

Inactive Publication Date: 2009-11-25
广西中医学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, that part of the extract of the white back leaf has the effect of inhibiting hepatitis virus and preventing and treating liver fibrosis, which is not disclosed in the open literature.

Method used

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  • Method of preparing extract of Mallotus apelta and uses for fighting hepatitis B virus
  • Method of preparing extract of Mallotus apelta and uses for fighting hepatitis B virus
  • Method of preparing extract of Mallotus apelta and uses for fighting hepatitis B virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Extraction and purification of flavonoids in the white-backed leaf plant:

[0029] Take the dry plant leaves of the white-backed leaves, remove impurities, crush or shred them, and extract in boiling water. Add 1-20L of water per kilogram of dry plant leaves, heat and extract for 1-8 hours, filter while hot, and collect the filtrate. The filter residue was extracted again with boiling water for 1 to 3 times, filtered while hot and then combined with the obtained filtrates.

[0030] The obtained filtrate is passed through a macroporous adsorption resin column at a temperature of 20°C to 80°C, eluted with water, and then eluted with 10% to 80% ethanol, and the ethanol eluate that reacts positively with the hydrochloric acid-magnesium powder solution is collected , dried under reduced pressure, and recrystallized to obtain the final product.

[0031] The macroporous resin is D101 macroporous resin or AB-8 resin. Plug the bottom of the chromatographic column...

Embodiment 2

[0033] Example 2: Inhibition of the expression of HBsAg and HBeAg in HepG 2215 cells by the flavonoids of Alba dorsa leaves. 2.1 Reagents: Eagles MEM dry powder (product of Gibco, USA); fetal bovine serum (product of Hyclone Lab, USA); G-418 (product of Genticin); L-glutamine (Amresco subpackage); Kanamycin (Amresco subpackage) ; DMSO (product of Biotech); 0.25% trypsin (product of Hyclone Lab, USA); hepatitis B surface antigen test kit and hepatitis B e antigen test kit (enzyme-linked immunoassay, product of Shanghai Rongsheng Biotechnology Co., Ltd.).

[0034] 2.2 In vitro cell culture: 2215 cells were constructed by the Mount Sinai Medical Center in the United States and donated by the Institute of Pharmaceutical Biotechnology, Chinese Academy of Medical Sciences. MEM medium for cell culture containing 10% fetal bovine serum, 300 μg / ml glutamine, 380 μg / ml G-418, 750 μg / ml kanamycin, with 5% NaHCO 3 Adjust the pH value to 7.2-7.4. Cells were cultured in MEM medium contain...

Embodiment 3

[0052] Example 3: Inhibition of DHBV-DNA replication by the flavonoids of the white back leaf

[0053] 3.1 Reagents: Positive control drug lamivudine (lamivudine, 3TC, GlaxoSmithKline Pharmaceutical Company); DMSO (Guangzhou Xingang Chemical Co., Ltd.); α-32P-Dctp (Beijing Furui Biotechnology Engineering Company); gap translation Kit (Promega); SephadexG-50 (Ficoll); PVP (Pharmacia); SDS (Merck); 0.45 μm nitrocellulose membrane (Amersham); protist DNA and bovine serum albumin were purchased from China Institute of Biophysics, Academy of Sciences.

[0054] 3.2 Experimental animals and source of virus: 1-day-old Peking duck, weighing 60-100 g, purchased from Beijing Qianjin Duck Farm. DHBV-DNA strong positive serum collected from Nanjing shelduck and stored at -70°C.

[0055] 3.3 Experimental method: 1-day-old Peking ducks were injected with 0.2ml of DHBV-DNA positive serum through the tibial vein of the legs per duck. Ducklings were randomly divided into 4 groups on the 7th ...

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Abstract

The invention relates to extract activity of Mallotus apelta and uses in prepartion of medicine for treating related diseases caused by hepatitis B. The extract of Mallotus apelta provided by the invention has basic components of apiolin, apiin-7-O-belta-D-glucoside, and flavonoids compounds of 5,7-dihydroxy-6-isopentene group-4'-methoxyl flavonone, flavonoids compounds Mallotus apelta have obvious inhibiting effect on hepatitis B virus surface antigen and hepatitis B virus e antigen ono surface of HepG 2215, and cabaple of inhibiting duplication of Duck Hepatitis B Virus deoxyribonucleic acid, withdraw rebound is weaker than positive reference medicine lamivudine, and hepatitis B is provided with a subsequent inhibiting effect after withdraw. The medicine according to the invention has strong function for fighting hepatitis B virus, clear active ingredient, which is suitable for industrial production, predictablly used for preparing medicine of preparing diseases infected by hepatitis B virus.

Description

technical field [0001] The present invention relates to an anti-hepatitis B virus Chinese medicine extract and its preparation method, in particular to the use of the anti-hepatitis B virus active agent of the active ingredient group obtained from the extraction of white back leaves to prepare drugs with the effect of treating various diseases caused by hepatitis B virus infection . Background technique [0002] Hepatitis B virus (HBV) is the pathogen of viral hepatitis B, and persistent HBV replication leads to liver inflammatory disease and causes hepatitis B. The disease is a common disease and frequently-occurring disease in our country, which has the characteristics of strong infectivity, complicated transmission route, wide epidemic area and high incidence rate. According to statistics from the Ministry of Health, the recorded cases of hepatitis B reached 910,000 in 2004 alone, and chronic asymptomatic HBV carriers may account for 9.09% of the general population, reac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7048A61K31/352A61P31/20A61P1/16A61K36/47A61K127/00
Inventor 郑作文韦松邓家刚
Owner 广西中医学院
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