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Medicinal herb solvent extract for treating hepatic disease

A technology of medicinal plants and extracts, applied in antiviral agents, drug combinations, plant raw materials, etc., can solve problems such as hepatitis B virus mutation and side effects

Active Publication Date: 2009-02-04
BELX BIO PHARMA (TAIWAN) CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Interferon was approved by the FDA for the treatment of chronic hepatitis B in 1992, but it will cause very large side effects during use, and only about 20% of liver B patients respond to interferon, and Ganan can be approved in 1998 Approved by the FDA to treat chronic hepatitis B, but only 17-33% of patients respond, and Chinese people have even lower response to Gananneng, and the worst is that long-term use of Gananneng can cause hepatitis B virus The mutation rate increases with the prolongation of use time, about 24% in the first year of use, 42% in the second year, 52% in the third year, and 52% in the fourth year as high as 67%
Therefore, it can be seen that so far, there is still no drug for the treatment of hepatitis B with high efficacy and low side effects and will not cause viral mutations

Method used

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  • Medicinal herb solvent extract for treating hepatic disease
  • Medicinal herb solvent extract for treating hepatic disease
  • Medicinal herb solvent extract for treating hepatic disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1: the extraction of medicinal plant extract

[0068] The extraction method of medicinal plant extract is as follows:

[0069] (1) Select Urticaceae plants of mountain ramie, long-leaf ramie, biting cat, water hemp, ramie, wood ramie.

[0070] (2) Wash the roots or stems of the nettle plant medicinal material in step (1), scrape off the outer skin, cut into thin slices, and grind them into powder.

[0071] (3) Get 600g of the powder in step (2) respectively, carry out reflux heating with 3000ml of 95% alcohol for 10 hours, filter and collect the pure liquid, concentrate under reduced pressure to remove ethanol, add ethyl ethyl ester and distilled water each 1000ml, After stirring for 30 minutes, it was left to stand for 1 hour.

[0072] (4) Extract the aqueous layer product of step (3) with 500 ml of ethyl acetate, and freeze-dry and concentrate the aqueous layer to obtain the extract.

Embodiment 2

[0073] Embodiment 2: Cell test of medicinal plant extract

[0074] The cell tests of medicinal plant extracts are as follows:

[0075] (1) The cell line HepG2.2.15 containing hepatitis B virus was selected and cultured in DMEM medium.

[0076] (2) Add 500 μg / ml of the extract of Example 1 to the cell line in step (1) and culture it for five days.

[0077] (3) On the first, third, and fifth days, the cell culture fluid in step (2) was collected and centrifuged (2000×g for 2 minutes).

[0078] (4) The centrifuged supernatant of step (3) is subjected to the test of hepatitis B virus surface antigen and e antigen inhibition; the centrifuged lower layer of cells is subjected to the test of cytotoxicity or hepatitis B virus DNA content. After the cells were washed twice with buffer solution, add 50 μl of MTT prepared in advance in DMEM culture medium and put it in 37°C culture medium to react for 30 minutes to 2 hours, then add 150 μl of DMSO to make the reaction color, and read t...

Embodiment 3

[0083] Embodiment 3: Urticaceae plant ITS sequence identification of medicinal plants

[0084] (1) The roots and stems of the Urticaceae plant medicinal materials are cleaned, scraped off the outer skin to reduce microbial contamination, then cut into thin slices, frozen and brittle with liquid nitrogen, and then ground into powder;

[0085] (2) The DNA extraction method mainly uses CTAB, PVPP and different salt concentrations, and is obtained through precipitation, centrifugation, Chloroform extraction and alcohol precipitation;

[0086] (3) Design the primer pair (primer) for PCR amplification amplification ITS sequence as follows:

[0087] 5'-CACACCGCCCGTCGCTCCTACCGA-3'

[0088] 5'-ACTCGCCGTTACTAGGGGAA-3'

[0089] Perform polymerase chain reaction at Tm=60°C;

[0090] (4) After the copied DNA fragments are analyzed by an automatic sequence analyzer (ABI 3100), the sequence similarity is compared with DNAMAN⑧ software;

[0091] Wherein the extraction step of the step (2)...

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Abstract

The invention relates to a solvent extract of uricaceous plant root or stem for hepatic disease treatment. The uricaceous plant has a ribosome intergenic sequence similarity greater than 70% with mountain ramie, ramie and longleaf ramie. In addition, The medical plant extract for hepatic disease treatment has an inhibitory effect to the wild type hepatitis B virus and the hepatitis B virus with lamivudine resistance.

Description

technical field [0001] The present invention relates to a kind of medicinal plant extract for liver disease treatment which can inhibit the replication of hepatitis B virus DNA and the secretion of hepatitis B virus surface antigen (HBs) and e antigen (HBe); the extract is not only effective for wild type ( Wild type) hepatitis B virus has an inhibitory effect, and it also has an inhibitory effect on viral DNA replication and secretion of viral surface antigen and e antigen of lamivudine-resistant hepatitis B virus. The source of the medicinal plant used for the treatment of liver disease is the Urticaceae plant with a specific ribosomal DNA internal transcribed spacer (ITS sequence for short), and the ITS sequence can be used as an identification comparison of the medicinal plant. Background technique [0002] There are 350 million people in the world who are chronic hepatitis B carriers. In the United States, there are about 1.25 million people with chronic hepatitis infec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K36/185C07H21/04A61P31/12A61K125/00A61K31/7088A61P1/16C07H1/08C12Q1/68
Inventor 李连滋张秀凤李承榆邱淑娇童天送
Owner BELX BIO PHARMA (TAIWAN) CORP
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