Anti-hepatitis B virus streblus extract and extracting technique thereof

Abstract

The invention discloses a streblus indica bur extract with hepatitis B virus resistance and a relative extraction method, wherein the basic formula of the extract is C3-C6-C6-C3 as streblus indica bur lignan compound. And the extraction method comprises breaking streblus indica bur, extracting via alcohol, methanol or water, extracting concentrated solution, extracting the concentrated solution via ligarine, ether, acetic ester and butanol respectively, processing silica gel column chromatography on the extracts, gradually eluting via at least one of ligarine, chloroform, acetic ester, ether and methanol, and obtaining streblus indica bur lignan compound. When the drug dosage is 100ug / ml, the inhibition rate on the HBsAg expression of hepatitis B virus cell line reaches at least 76.6% in vitro experiment, and when the drug dosage is 100ug / ml, the inhibition rate on the HbeAg expression reaches at least 81.6%. The invention discloses a new Chinese medicine source with hepatitis B virus resistance, with simple and mature extraction method and high value for developing high-effect low-toxin hepatitis B virus resistant drug.

Description

Technical field:

[0001] The invention relates to a traditional Chinese medicine and an extraction method, in particular to an anti-hepatitis B virus magpie kidney extract and an extraction process thereof. Background technique:

[0002] Hepatitis B is an infectious disease that seriously endangers human health. The pathogen that causes the disease is hepatitis B virus. At present, the medicines for treating hepatitis B at home and abroad mainly use interferon and nucleoside drugs, but these medicines are expensive, and the therapeutic effect is only about 30%, and after stopping the medicine, it is easy to produce recurrence. my country is a high-incidence area of ​​hepatitis B. The current prevalence rate is 277 people / 10,000 people, and the annual incidence rate is 95 people / 10,000 people. About 130 million people in the country are carriers of hepatitis B virus. The task of preventing and treating the disease is heavy and urgent. At present, more and more scientific and ...

Examples

Embodiment 1

[0017] (1) Extraction: Get 2 kilograms of naturally air-dried magpie kidney branches and leaves, heartwood, root, and bark, and use 75% ethanol to reflux after crushing, (3 × 30 liters), combine the extracts, recycle the solvent after filtration, and obtain red 1000 ml of the concentrated solution was placed in a 5000 ml separating funnel and extracted with petroleum ether, diethyl ether, ethyl acetate and n-butanol respectively. After recovering the solvent, 0.8 g of petroleum ether extract, 3.2 g of diethyl ether extract, acetic acid 33 grams of ethyl ester extract, 38 grams of n-butanol extract, and 87 grams of water-soluble matter.

[0018] (2) Separation:

[0019] Take 3.2 g of the ether extract and carry out silica gel column chromatography (300-400 mesh, 200 g), and use MeOH:CHCL 3 (0:100→5:95) gradient elution, five components were obtained, the weights were 0.3g, 0.6g, 0.5g, 0.2g and 1.2g, and each component was subjected to preparative thin-layer chromatography , e...

Embodiment 2

[0023] The effect of n-butanol extract of magpie kidney tree on the expression of HepG2.2.15, cell lines HBsAg and HbeAg:

[0024] Inoculate HepG2.2.15 cells 2×105 / ml into 96-well cell culture plate, 190ul per well, add the drug concentration of 800ug / ml, 400ug / ml, 200ug / ml, 100ug / ml into the cell culture plate, each concentration Set 3 wells, set 10% DMSO group, 3 wells are used as negative control, take the medium supernatant on the 3rd day and 6th day respectively, and store it at -20°C, and use the ELISA method to measure HBsAg and HbeAg at a wavelength of 450nm The absorbance at the place, the inhibition rate of the drug on the cells was calculated according to the following formula.

[0025]

[0026] See Table 1 and Table 2 for the results of the inhibition rate of the drug on the secretion of HbsAg and HbeAg:

[0027] Table 1 Suppressive effect of Que Kidney tree extract on HepG2.2.15 cell line HbsAg expression (X±D, n=3)

[0028]

[0029] The inhibitory ...

Technical field:

[0001] The invention relates to a traditional Chinese medicine and an extraction method, in particular to an anti-hepatitis B virus magpie kidney extract and an extraction process thereof. Background technique:

[0002] Hepatitis B is an infectious disease that seriously endangers human health. The pathogen that causes the disease is hepatitis B virus. At present, the medicines for treating hepatitis B at home and abroad mainly use interferon and nucleoside drugs, but these medicines are expensive, and the therapeutic effect is only about 30%, and after stopping the medicine, it is easy to produce recurrence. my country is a high-incidence area of ​​hepatitis B. The current prevalence rate is 277 people / 10,000 people, and the annual incidence rate is 95 people / 10,000 people. About 130 million people in the country are carriers of hepatitis B virus. The task of preventing and treating the disease is heavy and urgent. At present, more and more scientific and ...

Examples

Embodiment 1

[0017] (1) Extraction: Get 2 kilograms of naturally air-dried magpie kidney branches and leaves, heartwood, root, and bark, and use 75% ethanol to reflux after crushing, (3 × 30 liters), combine the extracts, recycle the solvent after filtration, and obtain red 1000 ml of the concentrated solution was placed in a 5000 ml separating funnel and extracted with petroleum ether, diethyl ether, ethyl acetate and n-butanol respectively. After recovering the solvent, 0.8 g of petroleum ether extract, 3.2 g of diethyl ether extract, acetic acid 33 grams of ethyl ester extract, 38 grams of n-butanol extract, and 87 grams of water-soluble matter.

[0018] (2) Separation:

[0019] Take 3.2 g of the ether extract and carry out silica gel column chromatography (300-400 mesh, 200 g), and use MeOH:CHCL 3 (0:100→5:95) gradient elution, five components were obtained, the weights were 0.3g, 0.6g, 0.5g, 0.2g and 1.2g, and each component was subjected to preparative thin-layer chromatography , e...

Embodiment 2

[0023] The effect of n-butanol extract of magpie kidney tree on the expression of HepG2.2.15, cell lines HBsAg and HbeAg:

[0024] Inoculate HepG2.2.15 cells 2×105 / ml into 96-well cell culture plate, 190ul per well, add the drug concentration of 800ug / ml, 400ug / ml, 200ug / ml, 100ug / ml into the cell culture plate, each concentration Set 3 wells, set 10% DMSO group, 3 wells are used as negative control, take the medium supernatant on the 3rd day and 6th day respectively, and store it at -20°C, and use the ELISA method to measure HBsAg and HbeAg at a wavelength of 450nm The absorbance at the place, the inhibition rate of the drug on the cells was calculated according to the following formula.

[0025]

[0026] See Table 1 and Table 2 for the results of the inhibition rate of the drug on the secretion of HbsAg and HbeAg:

[0027] Table 1 Suppressive effect of Que Kidney tree extract on HepG2.2.15 cell line HbsAg expression (X±D, n=3)

[0028]

[0029] The inhibitory ...

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