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Method for extracting medicament for killing hepatitis C virus from garlic

A technology for extracting hepatitis C virus and garlic, which is applied in antiviral agents, drug combinations, and pharmaceutical formulas, can solve the problems of no hepatitis C virus drugs, and achieve the effects of low cost, simple method, and cheap raw materials

Inactive Publication Date: 2010-12-29
姚传凤
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But then there is no report for being able to kill hepatitis C virus from garlic; There is no report for extracting the material of killing hepatitis C virus contained in garlic with what method
After retrieving domestic and foreign patent documents and consulting public publications at home and abroad, there is no report on the method of extracting and killing hepatitis C virus medicine from garlic

Method used

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  • Method for extracting medicament for killing hepatitis C virus from garlic

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Determination of the cytotoxicity of the extract to Huh-7.5.1

[0027] Digest Huh-7.5.1 cells with trypsin, press 2×10 4 Inoculated into 96-well cell culture plates at a density of 5% CO 2 Incubate in an incubator at 37°C for 6 hours. After the cells adhere to the wall, add 10 μl of various extracts of different concentrations to each well, culture overnight at 37°C, add 10 μl of CCK-8 reagent to each well, and continue to incubate for 4 hours. At the same time, set up a negative control, add CCK-8 to the cell wells without extracting solution; add CCK-8 to the blank control, without cells, add DMEM culture solution. The 450nm absorbance value (OD value) of each well was measured with a microplate reader.

Embodiment 2

[0029] Antiviral activity of garlic extract:

[0030] In order to screen whether the extract has anti-HCV activity, as well as the pathway of antiviral effect, the following three groups of parallel experiments were designed:

[0031] The first group: kill the virus:

[0032] Take 3 concentrations: 10 μl each of garlic extracts of 20mg / ml, 5mg / ml, and 1mg / ml, and first mix them with 10μl HCVcc in equal amounts, and then inoculate them into 96-well plates after acting at 37°C for 4 hours. The cell density was 3 x 10 4 / hole, the same below. After culturing for 72 hours, the IFA experiment was carried out to detect the expression of HCV protein in the cells. A well directly infected with HCVcc was used as a positive control, and a well without extract and virus was used as a negative control.

[0033] The second group: blocking virus infection:

[0034] First add 10 μl of different concentrations of extracts to a 96-well plate containing Huh-7.5.1 cells, culture overnight...

Embodiment 3

[0038] IFA detection: Aspirate the culture supernatant in the 96-well plate, add 100 μl of methanol to each well, fix at -20°C for 20 minutes, wash with PBS 3 times for 3 minutes each time. Add 100 μl of 0.1% TritonX-100( ) prepared in PBS to each well, permeabilize at room temperature for 30 min, and wash with PBS (3 min×3 times). Add 100 μl of blocking solution (PBS containing 3% BSA) to each well, and block for 1 hour at room temperature. The blocking solution was sucked off; 50 μl of hepatitis C patient serum was added to each well (the serum diluted 1:50 with the blocking solution was used as the primary antibody), incubated at room temperature for 2 hours, and washed 3 times with PBS, 10 minutes each time. Fluorescein FITC-labeled anti-human IgG was added to each well as the secondary antibody, incubated in the dark for 1 hour, and washed with PBS (3 min×3 times).

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Abstract

The invention belongs to medicament preparation methods, in particular to a method for extracting a medicament for killing hepatitis C viruses from garlic. The method comprises the following steps of: accurately weighing 20 parts by weight of garlic, pulverizing to 100 meshes, and adding 110 parts by weight of 95-percent ethanol for soaking for 2 hours; carrying out hot reflux for 4 hours on a sample at the temperature of 80 DEG C through a Soxhlet extractor; filtering a solution; concentrating the filtered solution to 20 parts by weight through a rotary evaporator; and sterilizing the concentrated solution through filtration, and diluting through a DMEM (Dulbecco's Modified Eagle Medium) culture fluid. The invention has the advantages of cheap raw materials, low cost, simple method, convenient extraction and better killing effect for the hepatitis C viruses.

Description

technical field [0001] The invention belongs to a method for preparing medicine, in particular to a method for extracting and killing hepatitis C virus medicine from garlic. Background technique [0002] Hepatitis C is a disease that seriously endangers human health and is difficult to cure. Hepatitis C is a disease caused by infection of the human body with the hepatitis C virus. Due to the extremely strong replication ability of the hepatitis C virus, there is a hard shell on the surface of the virus, and it is difficult for ordinary drugs to penetrate into the virus body, so the virus is difficult to kill, and hepatitis C is also difficult to cure. The prior art is to treat with antiviral western medicine, but due to the particularity of the hepatitis C virus structure, it can only alleviate the symptoms of the patient in a short period of time, but cannot cure the disease. Therefore, it is an important task to invent a method for extracting and killing hepatitis C viru...

Claims

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Application Information

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IPC IPC(8): A61K36/8962A61P1/16A61P31/14A61K125/00
Inventor 姚传凤杨宇
Owner 姚传凤
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