Prunella vulgaris L. extract with anti-HSV activity, and preparation method and medical application of Prunella vulgaris L. extract

A technology of Prunella vulgaris and its extract, applied in the field of medicine, can solve problems such as easy generation of drug resistance, and achieve the effect of stable anti-HSV effect, good curative effect, and inhibition of hind limb paralysis

Active Publication Date: 2021-10-08
SHANGHAI UNIV OF T C M
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinical treatment drugs are mainly nucleoside analogues such as acyclovir (ACV). For severe and frequent relapse cases, large doses of long-term administration are required, which is prone to drug resistance

Method used

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  • Prunella vulgaris L. extract with anti-HSV activity, and preparation method and medical application of Prunella vulgaris L. extract
  • Prunella vulgaris L. extract with anti-HSV activity, and preparation method and medical application of Prunella vulgaris L. extract
  • Prunella vulgaris L. extract with anti-HSV activity, and preparation method and medical application of Prunella vulgaris L. extract

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Technical scheme of the present invention carries out according to the following steps:

[0042] The preparation method of different extracting parts of Prunella vulgaris comprises the following steps:

[0043] (1) Preparation of different solvent extracts of Prunella vulgaris (see figure 1 )

[0044] Prunella vulgaris 90% ethanol extract: Take 50 g of Prunella vulgaris, add 20 times the volume and mass percentage of ethanol with a concentration of 90% to extract twice, filter for 2 hours, discard the drug residue, concentrate the extract, and concentrate under reduced pressure by rotary evaporation at 50 ° C. After drying, the 90% ethanol extract of Prunella vulgaris was obtained, which was named PV90E.

[0045] Prunella vulgaris 70% ethanol extract: take 50 g of Prunella vulgaris, add 20 times the volume and mass percentage of ethanol with a concentration of 70% to extract twice, filter for 2 hours, discard the drug residue, concentrate the extract, and concentrate ...

Embodiment 2

[0055] In this example, chemical characterization was carried out by using differential reaction, HPGPC and graded alcohol precipitation parts (PVE30, PVE50, PVE70, PVE85), and the ethanol extracts of Prunella vulgaris (PV90E, PV70E) and water extraction alcohol precipitation supernatant (PVES) UPLC-DAD-QTOF-MS / MS analysis was performed.

[0056] (1) The identification reaction of each part of Prunella vulgaris water extract, alcohol extract and graded alcohol precipitation

[0057] Method: Molish reaction (sugars), iodine solution reaction (starch), Coomassie brilliant blue reaction (protein), ninhydrin reaction (free amino acid) and FeCl reaction were carried out on the above parts. 3 Reaction (tannins), acetic anhydride concentrated sulfuric acid reaction (saponins), Feigl reaction (quinones), magnesium hydrochloride powder reaction (flavonoids), potassium iodide precipitation reaction (alkaloids), etc.

[0058] Results: The Molish reaction, ferric chloride reaction, aceti...

Embodiment 3

[0087] The anti-HSV-1 / KOS, HSV-2 / G standard strain activity test was carried out on each extract part of Prunella vulgaris prepared in Example 1, and the part with the optimal anti-HSV activity was screened out.

[0088] Method: CCK-8 colorimetric method was used to detect the in vitro inhibitory effect of Prunella vulgaris extract on standard virus strains HSV-1 / KOS and HSV-2 / G.

[0089] After counting the Vero cells, they were seeded in a 96-well cell culture plate at a density of 20,000 cells per well, and placed in a constant temperature of 37°C, CO 2 In an incubator with a concentration of 5% by volume, after the cells grow to a monolayer, discard the upper layer of culture solution, and add 100×TCID 50 After one hour of adsorption, different concentrations of Prunella vulgaris extract samples (100 μg / mL, 50 μg / mL, 25 μg / mL, 12.5 μg / mL) were added, and each concentration was repeated in 3 wells, 200 μL / well, at 37 ℃, volume percentage 5% CO 2 Cultivate for 3 days, add C...

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Abstract

The invention relates to a Prunella vulgaris L. extract with an anti-HSV activity. The Prunella vulgaris L. is prepared from the following steps of: taking Prunella vulgaris L. clusters or powders, adding 10-22 volume times of water, carrying out heating, refluxing and extracting for 2-4 times, filtering, blending filtrate, concentrating under reduced pressure, then adding 0.2-1 volume times of ethanol until the ethanol concentration is 10-50% v/v, standing for 6-24 hours at 4 DEG C, centrifuging, washing the obtained precipitate with the ethanol of which the concentration is 10-50% v/v for at least two times, collecting the precipitate, and drying. The invention also provides a preparation method and application of the Prunella vulgaris L. extract. The Prunella vulgaris L. extract provided by the invention has a remarkable in-vivo and in-vitro anti-HSV effect, the in-vitro anti-HSV activity of the Prunella vulgaris L. extract is increased by one time than the in-vitro anti-HSV activity of an aqueous extract, and the Prunella vulgaris L. extract also has a stable inhibition effect for ACV drug-resistant HSV clinical strains and drug-resistant strains. Meanwhile, the Prunella vulgaris L. extract has a good curative effect for skin and genital herpes caused by HSV-1 and HSV-2 infection.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to a Prunella vulgaris, in particular to a Prunella vulgaris extract with anti-HSV activity, a preparation method and a medical application thereof. Background technique [0002] Herpes simplex virus (HSV) is divided into type I (HSV-1) and type II (HSV-2). The virus invades the body through the oral cavity and genitals. disease and serious complications. Clinical treatment drugs are mainly nucleoside analogues such as acyclovir (ACV). For severe and frequent relapse cases, large doses of long-term administration are required, and drug resistance is prone to occur. Therefore, there is an urgent need to develop anti-HSV drugs with different structures and mechanisms of action from acyclovir. [0003] The applicant team established a traditional Chinese medicine anti-virus screening platform in the early stage, and systematically screened traditional Chinese medicines with heat-clear...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/536A61P31/22A23L33/105A61K131/00
CPCA61K36/536A61P31/22A23L33/105A61K2236/331A61K2236/39A61K2236/53A61K2236/51A23V2002/00A23V2200/30A23V2250/21
Inventor 徐宏喜张群铄李洋付文卫钟煊蕾冯极灵罗小妹袁满
Owner SHANGHAI UNIV OF T C M
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