CN103446110A - Applications of Sarcaboside A in medicament against herpes virus - Google Patents

Abstract

The invention discloses applications of Sarcaboside A in preparing a medicament against herpes virus. Experiments prove that the Sarcaboside A plays a remarkable role in inhibiting Herpes simplex virus HSV-1 and Herpes simplex virus HSV-2, thus indicating that the Sarcaboside A has potential application values in the field of treating herpes virus-infected diseases. The applications provide an experimental basis for the Sarcaboside A in clinically treating herpes virus-infected diseases, and offer certain guiding significance for developing medicaments against HSV-1 and HSV-2, thus having an important reference value.

Description

The application of Sarcaboside A in the medicine of anti-herpesvirus

Technical field

The present invention relates to the application of Sarcaboside A in preparing anti-herpes simplex virus 1 type (Herpes simplex virus, HSV-1) and herpes simplex virus type 2 (Herpes simplex virus, HSV-2) medicine.

Background technology

HSV-1, HSV-2 belong to the herpes coe virus, and HSV-1 and HSV-2 not only cause lip or reproduction herpes mucosae, are also to cause the even important pathogen of intrauterine infection of encephalitis, hepatitis, whole body severe infections simultaneously;

The compound S arcaboside A the present invention relates to is one and within 2012, delivers (Li, X.et al., 2012.Two New-Skeleton Compounds from Sarcandra glabra.Helvetica Chimica Acta95 (6), 998-1002.) New skeleton compound, this compound has brand-new framework types, at present not about active report, purposes for the Sarcaboside A the present invention relates in preparation treatment herpesvirus infection medicine belongs to open first, because framework types belongs to brand-new framework types, and it suppresses active unexpectedly strong for herpesvirus, there do not is the possibility that is provided any enlightenment by other compounds, possesses outstanding substantive distinguishing features, control for herpesvirus infection simultaneously obviously has significant progress.

Summary of the invention

For above-mentioned prior art, the invention provides the new purposes of Sarcaboside A in pharmacy:

The application of Sarcaboside A in the medicine of the anti-HSV-1 of preparation.

The application of Sarcaboside A in the medicine of the anti-HSV-2 of preparation.

The experiment proved that, Sarcaboside A has significant inhibitory action to HSV-1, HSV-2: suppress HSV-1, HSV-2 and CPE effect (Cytopathic Effect, CPE), IC occur at the HEK293 cell ₅₀be respectively 2 ^-12.11, 2 ^-11.44, TI is respectively 1807.77,1136.20.

Described compound S arcaboside A structure is as shown in formula I:

The present invention is used for the treatment of the herpesvirus infection disease for Sarcaboside A is clinical provides experimental basis, to the drug provision of developing anti-HSV-1, HSV-2 virus certain directive significance, there is important reference value.

The purposes of the Sarcaboside A the present invention relates in preparation treatment herpesvirus infection medicine belongs to open first, because framework types belongs to brand-new framework types, and it suppresses active unexpectedly strong for herpesvirus, there do not is the possibility that is provided any enlightenment by other compounds, possess outstanding substantive distinguishing features, the control for yellow herpesvirus infection simultaneously obviously has significant progress.

In order to understand better essence of the present invention, below in conjunction with embodiment, the present invention is further illustrated.

The specific embodiment

The preparation method of compound S arcaboside A involved in the present invention is referring to document (Li, X.et al., 2012.Two New-Skeleton Compounds from Sarcandra glabra.Helvetica Chimica Acta95 (6), 998-1002.).

The present invention is further detailed explanation by the following examples, but protection scope of the present invention is not subject to any restriction of specific embodiment, but be limited by claim.

Embodiment 1: the preparation of compound S arcaboside A tablet involved in the present invention:

Get 20 and digest compound Sarcaboside A, add conventional adjuvant 180 grams that prepare tablet, mix, conventional tablet machine is made 1000.

Embodiment 2: the preparation of compound S arcaboside A capsule involved in the present invention:

Get 20 and digest compound Sarcaboside A, add the conventional adjuvant for preparing capsule as starch 180 grams, mix, encapsulatedly make 1000.

Further illustrate its pharmaceutically active below by pharmacodynamic experiment.

Experimental example: the effect research of Sarcaboside A to herpesvirus:

(1) cell strain and Strain:

HEKC (HEK293 is HSV-1, HSV-2 virus sensitive cells) is purchased from U.S. Clontech company; Herpes simplex virus type 1 (HSV-1) Sm44 strain, simple herpes virus 2 types (HSV-2) 333 strains are drawn from Chinese CDC virosis institute.

(2) cytotoxic assay:

Reference literature (Wang Shouchuan, Wang Lin, old superfine, the experimentation of oral liquid for clearing away lung-heat Contained Serum to viral inhibition, Nanjing University of Traditional Chinese Medicine's journal, 2008; 24(1): method 25～27), each sample is done to serial dilution (2 with 2 multiple proportions ^0～-5), then be inoculated in HEK293 in 96 plate holes by the dilution sequential lateral upper, every hole 100uL, every dilution factor vertically repeats 3 holes (A, B, C capable), parallel 1～6 hole of establishing microwell plate D is the cell contrast, 7～12 holes not inoculating cell are blank, and microscope is observed CPE, Continuous Observation 7d lower every day, neutral red staining, measure the OD value at the 540nm wavelength, experimental group is compared to the calculating cell survival rate with cell matched group OD value, by the Reed-Muench method, calculate medicine half cytotoxic concentration (TD ₅₀).

(4) toxin inhibitory test:

Reference literature (Wang Shouchuan, Wang Lin, old superfine, the experimentation of oral liquid for clearing away lung-heat Contained Serum to viral inhibition, Nanjing University of Traditional Chinese Medicine's journal, 2008; 24(1): method 25～27), sample is pressed to dilution order (2 ^-3～-14) laterally be inoculated on the cell monolayer in 96 plate holes, every hole 100uL, every dilution factor vertically repeats 4 holes, and the A of microwell plate, B, C is capable adds containing 100 TCID ₅₀viral 100uL as test group, D is capable, and the capacity cell maintenance medium that supplement to wait is the contrast of variable concentrations medicine, the parallel F that establishes is capable of virus control, what E was capable 1～12 contrasts as cell.37 ℃, 5%CO ₂cultivate, observe CPE every day, Continuous Observation 4d.When 90% above CPE appears in virus control, add 1% neutral red staining, by microplate reader, at wavelength 540nm wavelength, read the A value, each is organized the A value and removes virus control group A value, each test group is compared with cell control group A value, obtained cell survival rate, calculate half by the Reed-Muench method and press down malicious concentration (IC ₅₀), TD the most at last ₅₀and IC ₅₀compare to obtain and press down malicious index (TI).

(5) result:

1. sample TD ₅₀mensuration: the micro-Microscopic observation adherent growth of cellular control unit is fine and close, form is good.The TD of SarcabosideA ₅₀be respectively 2 at HEK293 ^-1.29.

2. press down the poison experiment: the adherent densification of cellular control unit, form are good.Microscopic observation is found, HSV-1, HSV-2 virus control group the CPE more than 90% occurs at 3d, 3d respectively, HSV1 and HSV2 HEK293CPE with swelling, many cells merge (HSV1 causes that a plurality of cells are warm becomes bulla, and HSV2 cause be fused to vesicle), become circle, come off into principal character.

And toxin inhibitory test is respectively organized cell; according to Sarcaboside A diluted sample gradient; the gradient rule occur CPE:HSV1-HEK293 before the 10th dilution factor, HSV2-HEK293 group cell before the 9th dilution factor is fully protected, CPE occurs afterwards, CPE reduces and increases the weight of gradually with sample concentration.

By 1% neutral red staining for above-mentioned brassboard, by microplate reader, survey 450nmA value, after each is organized the A value and cuts virus control group A value, each experimental group A value is compared acquisition IC with cell control group A value ₅₀, IC ₅₀with TD ₅₀compare the TI that obtains Sarcaboside A: suppress HSV-1, HSV-2, at the HEK293 cell, CPE, IC occur ₅₀be respectively 2 ^-12.11, 2 ^-11.44, TI is respectively 1807.77,1136.20.

Conclusion: Sarcaboside A has the effect of (comprising HSV-1, HEV-2) of significant herpes coe virus, and safe, so Sarcaboside A has a wide range of applications background in treatment herpesvirus infection disease.

Claims (1)

1.Sarcaboside the application of A in the medicine of anti-herpesvirus, described compound S arcaboside A structure as formula Ishown in:

formula I.