CN113876765A

CN113876765A - Application of Werethrin in the preparation of medicines and health products for preventing and/or treating herpes virus infection

Info

Publication number: CN113876765A
Application number: CN202111261751.5A
Authority: CN
Inventors: 郝翠; 王伟; 王兆琦; 姚如永
Original assignee: Ocean University of China; Affiliated Hospital of University of Qingdao
Current assignee: Ocean University of China; Affiliated Hospital of University of Qingdao
Priority date: 2021-10-28
Filing date: 2021-10-28
Publication date: 2022-01-04

Abstract

The invention discloses application of wedelolactone in preparation of a medicine and a health-care product for preventing and/or treating herpes virus infection. Experiments prove that wedelolactone has an activity inhibiting effect on herpes simplex virus infection, namely wedelolactone has the activity of resisting HSV-1 and HSV-2, and is low in toxicity. Wedelia lactone can obviously inhibit related protein and mRNA expression of HSV-1 and HSV-2, and both have obvious dose dependence, and have the best effect in the early stage after HSV infection, namely 2-4 h. Wedelolactone can directly inactivate virus particles or intervene the replication process of viruses after entering cells, inhibit virus proliferation to block herpes virus infection, and can reduce the virus load in mouse tissues at the level of in vivo animals. Therefore, wedelolactone has the potential to be developed into a novel anti-HSV preparation of the targeted virus particles.

Description

Application of wedelolactone in preparation of medicines and health products for preventing and/or treating herpes virus infection

Technical Field

The invention belongs to the field of medicines, and particularly relates to application of wedelolactone in preparation of medicines and health-care products for preventing and/or treating herpes virus infection.

Background

Herpes virus is an enveloped double-stranded DNA virus belonging to the family of herpesviridae, most typically herpes simplex virus, and is mainly classified into type 1 (HSV-1) and type 2 (HSV-2). Infections are very common in people, 80% -90% of primary infections are recessive infections and have no obvious symptoms, wherein HSV-1 infection causes skin injury and is generally limited to oral, nasal and ocular levels, while for HSV-2, the infection is most common in genital skin, mucous membranes and other parts. Current FDA-approved antiviral agents for herpes viruses are primarily directed to nucleoside analogs, such as Acyclovir (ACV) and penciclovir, which primarily inhibit viral genome replication. Despite these successes, drug resistance and side effects have not solved the problem of HSV infection. Therefore, the development of novel anti-HSV agents with mechanisms of action different from nucleoside analogues is of great clinical significance.

Wedelolactone (C)₁₆H₁₀O₇) Is prepared from wedelia chinensis (Wedelia chinensis)Wedelia chinensis) Yerbadetajo herb (Yerbadetajo herb)Eclipta prostrataL.) coumarin compound extracted from the above-mentioned raw materials. Wedelolactone has various biological activities, and has pharmacological effects of protecting liver, resisting immunosuppression, resisting inflammation, promoting bone differentiation, and resisting cancer. However, the effect of wedelolactone on a simple viral infection has not been reported.

Disclosure of Invention

The invention provides application of wedelolactone in preparation of a medicine and a health-care product for preventing and/or treating herpes virus infection. Experiments verify that wedelolactone has an inhibition effect on HSV-1 and HSV-2, is extremely low in toxicity, and has the potential of being developed into novel anti-HSV drugs and health care products.

In order to realize the purpose of the invention, the invention is realized by adopting the following technical scheme:

the invention provides application of wedelolactone in preparation of a medicine and a health-care product for preventing and/or treating herpes virus infection.

Further, the herpes virus is herpes simplex virus, specifically HSV-1 virus and HSV-2 virus.

Further, the wedelolactone is used at a concentration of 0.1-50 μ M.

Furthermore, the wedelolactone can effectively inhibit HSV-2 virus at a use concentration of 0.25-2 mu M.

Preferably, the wedelolactone is used for effectively inhibiting HSV-2 virus at a concentration of 0.5-2 mu M.

Furthermore, the wedelolactone can effectively inhibit HSV-1 virus at the use concentration of 1.25-10 mu M.

Preferably, the wedelolactone is used for effectively inhibiting HSV-1 virus at a concentration of 5-10 mu M.

Further, the wedelolactone is used at a concentration of 10 μ M with the best effect against herpes viruses.

Furthermore, the medicament and the health-care product are prepared by taking wedelolactone and pharmaceutically acceptable salts, stereoisomers, crystals or derivatives thereof as main active components and together with pharmaceutically auxiliary components.

Furthermore, the using time of the medicine and the health care product is within 2-6 hours after herpes virus infection.

Preferably, the using time of the medicine and the health care product is within 2-4 hours after herpes virus infection.

Furthermore, the dosage of the main active component wedelolactone in the medicine and the health care product is 2.5 mg/kg/d-5 mg/kg/d.

Preferably, the medicament and the health-care product contain wedelolactone as a main active component, and the dosage of the wedelolactone is 5 mg/kg/d.

Furthermore, the medicament and the health care product achieve the effect of blocking the infection of the herpesvirus by the direct action of the wedelolactone contained in the medicament and the health care product and the herpesvirus.

The invention also provides a medicament for preventing and/or treating herpes virus infection, which takes wedelolactone and pharmaceutically acceptable salts, stereoisomers, crystals or derivatives thereof as main active ingredients.

Compared with the prior art, the invention has the advantages and positive effects that:

the wedelolactone is selected as a research object, and the effect of the wedelolactone on inhibiting the herpes simplex virus is disclosed at the in vivo and in vitro levels. Experiments prove that the natural product wedelolactone has a good inhibition effect on HSV-1 and HSV-2 infection, and can inhibit related protein and mRNA expression of HSV-1 and HSV-2. Wedelolactone can interfere virus adsorption, improve herpes simplex virus infected mouse body condition, and inhibit virus proliferation in vivo. The wedelolactone is firstly determined to have the inhibition effect on HSV-1 and HSV-2 and has extremely low toxicity. Therefore, the wedelolactone can be possibly developed into a novel HSV (herpes Simplex Virus) resistant medicament, and has a good market application prospect.

Drawings

FIG. 1 is a diagram of the results of detection of the cytotoxicity of wedelolactone on Vero, Hep-2 and HeLa cells.

FIG. 2 shows the result of plaque experiment of wedelolactone entering Vero cell to inhibit HSV-1 and HSV-2 virus in different action modes.

Figure 3 is a plot of the results of a plaque reduction experiment in which wedelolactone has a direct inactivation effect on HSV.

Figure 4 is a graph of the inhibition of HSV by wedelolactone at different action times.

FIG. 5 is a graph showing the results of inhibition of ICP27 protein expression of HSV-1 virus by different concentrations of wedelolactone.

FIG. 6 is a graph of the results of inhibition of gB protein expression by different concentrations of wedelolactone on HSV-2 virus.

FIG. 7 is a graph of the results of inhibition of HSV-1 gD protein mRNA expression by wedelolactone.

Figure 8 is a graph of the effect of wedelolactone on the body weight of HSV-1 infected mice.

Figure 9 is a result graph of the effect of wedelolactone on survival of HSV-1 infected mice.

Figure 10 is a graph of the results of wedelolactone reducing viral load in HSV-1 infected mouse spinal cord tissue.

Detailed Description

The technical solution of the present invention will be described in further detail with reference to the accompanying drawings and the detailed description.

According to an internationally recognized method, the inhibition effect and mechanism of wedelolactone on herpes simplex virus are evaluated. The wedelolactone disclosed by the invention can be prepared from a commercially available product.

Example 1: wedelolactone for inhibiting herpes simplex virus in vitro

1. Experimental methods

1.1 cytotoxicity assay of Wedelia lactone

The cytotoxicity of wedelolactone on three different cells was determined. Respectively inoculating Vero, Hela and Hep-2 cells into a 96-well plate until the cells grow into a monolayer, removing the original culture solution, adding wedelolactone (with final concentration of 400 μ M, 200 μ M, 100 μ M, 50 μ M, 25 μ M, 12.5 μ M or 300 μ M, 150 μ M, 175 μ M, 37.5 μ M and 18.75 μ M) into the 96-well plate, wherein each concentration is three multiple wells, and a blank control group is arranged. The cells were incubated at 37 ℃ in 5% CO₂Culturing for 24h in a constant-temperature cell culture box, removing the drug, adding 4% paraformaldehyde, fixing at room temperature for 15min, removing the drug, adding 5% crystal violet, dyeing at room temperature for 15min, cleaning, air drying, and measuring A with an enzyme-labeling instrument_540nmThe value is obtained.

Cell viability test group a_540nmNegative control group A_540nm×100%。

1.2 Wedelia lactone virus inhibition experiment by detecting cytopathic effect (CPE)

Vero, Hela, Hep-2 cells in 96-well plates were infected with HSV-1 or HSV-2 with a multiplicity of infection (MOI) of 0.1. The wedelolactone is diluted according to gradient, and Vero, HeLa and Hep2 cells infected by HSV-1 and HSV-2 are subjected to whole-process administration (four modes of pretreating cells and viruses, adsorbing and then adsorbing). Pretreatment cells (predose + cells): wedelia lactone acts on cell 1 at 37 ℃h, absorbing and discarding, adsorbing HSV (MOI = 0.1) at 37 ℃ for 1h, and changing into a maintenance solution; pretreatment virus (predose + HSV): mixing wedelolactone and HSV at 37 deg.C, incubating for 1h, adding into 96-well plate with original culture medium, reacting at 37 deg.C for 1h, and changing into maintenance solution; administration during adsorption (adsorption dosing): mixing wedelolactone and HSV (MOI = 0.1), adding into 96-well plate, reacting at 37 deg.C for 1h, and changing into maintenance liquid; administration after adsorption (dosing after adsorption): HSV is used for adsorbing cells for 1h at 37 ℃ and is changed into maintenance liquid containing wedelolactone. Each concentration was set with 3 replicate wells, along with a virus control and a blank control. After 30h, test A according to 1.1_540nm。

Cell survival rate ═ test group a_540nmViral control group A_540nm) /(blank control group A)_540nmViral control group A_540nm）×100%。

1.3 detection of Wedelia lactone in Virus inhibition by plaque titer test

Vero cells were seeded into 12-well plates until they grew full of a monolayer. Wedelolactone (final concentration 10 μ M) was added to each of 4 different administration modes in the 1.2 experimental method, and after 24h, the supernatant was collected and a virus control group was set. Diluting the collected supernatant 10^-1~10^-3And adsorbing the cells at 37 ℃ for 1h, removing the cells by suction, adding the prepared overlay culture medium, cooling and solidifying the cells, and then inversely placing the cells in an incubator at 37 ℃. After spotting, the staining was fixed and the number of plaques was counted.

1.4 detection of the direct Virus-inhibiting Effect of Wedelia lactone in the plaque reduction assay

Vero cells were seeded into 12-well plates until they grew full of a monolayer. Mixing wedelolactone (10 μ M, 5 μ M, 2.5 μ M, 1.25 μ M, 0.625 μ M) and HSV-1 at 37 deg.C for 1h, adding into 12-well plate with original culture medium, allowing to act at 37 deg.C for 1h, removing, adding prepared overlay culture medium, cooling and solidifying, and placing in 37 deg.C incubator. After spotting, the staining was fixed and the number of plaques was counted.

2. Results of the experiment

2.1 cytotoxicity assay of Wedelia lactone

The experimental result is shown in figure 1, and it can be seen that the cell survival rate of Vero is close to 80% when the wedelolactone concentration is 50 μ M; at a concentration of 75. mu.M, HeLa showed a cell survival rate of approximately 80%, and at a concentration of 150. mu.M, Hep-2 showed a cell survival rate of approximately 80%. With the increasing concentration of wedelolactone, the cell survival rate gradually decreases. CC in Vero cells₅₀CC in HeLa cells at 175.6. mu.M₅₀CC at 227.4. mu.M in Hep-2 cells₅₀It was 250.0. mu.M.

2.2 evaluation of Wedelia chinensis lactone anti-HSV Activity in different cells by CPE inhibition experiment

CPE inhibition experiment evaluates the activity of wedelolactone in resisting HSV-1 and HSV-2 in Vero, HeLa and Hep-2 cells, and half Inhibition Concentration (IC) is calculated by the administration of the whole process₅₀) And calculating the therapeutic index (SI = CC)₅₀/IC₅₀). As shown in Table 1, wedelolactone has a very good anti-HSV effect on three cells, and IC thereof₅₀The values are all less than 5 μ M and have a certain dose dependence. The comparison shows that the inhibition effect of the wedelolactone has certain cell specificity, and the effect of the wedelolactone on resisting HSV-2 on Vero cells is particularly obvious.

TABLE 1 cytotoxic and antiviral effects of wedelolactone in different cells

2.3 evaluation of the mode of action of Wedelia lactone on the inhibition of HSV

As shown in fig. 2, wedelolactone acts on Vero cells by four modes of cell pretreatment, virus pretreatment, administration during adsorption and administration after adsorption. Compared with HSV-1 virus group, the virus titer is respectively reduced by 0.88, 5.9, 0.24 and 1.24 Log₁₀(PFU/mL); compared with HSV-2 virus group, the virus titer is respectively reduced by 0.12, 3.09, 0.13 and 1.91 Log₁₀(PFU/mL)。

2.4 plaque reduction test evaluation of the direct inactivation of HSV by Wedelia lactone

The experiments show that wedelolactone can reduce virus titer by directly interacting with HSV, so that we further verify whether wedelolactone can be inactivated by directly interacting with virus by using a plaque reduction test. As shown in figure 3, a plaque reduction test is more intuitive, and the wedelolactone has a direct inactivation effect on HSV, and can obviously inhibit the formation of plaques of HSV virus at a concentration of more than 0.625 mu M.

Example 2: mechanism of action of wedelolactone for resisting HSV

1. Experimental methods

1.1 Indirect immunofluorescence assay

Inoculating Vero cells into a 3.5 cm glass substrate, treating HSV-1 (MOI = 1) by wedelolactone (final concentration is 10 mu M) according to 4 different action modes in 1.2 in example 1 when the cells grow to the density of 30-40%, and culturing in an incubator at 37 ℃ for 8 hours; then, cells were washed with PBS, fixed with 4% paraformaldehyde at room temperature for 15min, washed with PBS for 2min, permeabilized with 0.25% Triton X-100 for 10min, washed with PBS for 2min, blocked with 2% BSA at 37 ℃ for 1h, washed with PBS three times, incubated with an antibody to ICP5 (1: 100 dilution) at 4 ℃ for 16h, washed, incubated with FITC-conjugated secondary antibody at 4 ℃ for 4h, washed three times, treated with DAPI for 10min, washed, confocal imaged, and analyzed using Image J (NIH).

1.2 Western blot assay

Inoculating Vero cells into a 6-well plate, mixing and pretreating wedelolactone, HSV-1 and HSV-2 in a gradient dilution mode for 1h at 37 ℃ after the cells grow over a monolayer, adding the mixture into the 6-well plate, adsorbing the cells for 1h at 37 ℃, removing the solution, replacing the solution with a maintenance solution, and meanwhile, arranging a virus control group and a blank control group. Vero cells are inoculated in a 12-hole plate, HSV-1 (MOI = 0.1) adsorbs the cells for 1h at 37 ℃, and wedelolactone (the final concentration is 40 mu M) is added in different time periods (0-2 h, 2-4h, 4-6h, 6-8h, 8-10h, 10-12h, 0-6h and 0-12 h) after adsorption.

After 16h, samples were collected, 100. mu.L of cell lysate (800. mu.L of RIPA, 200. mu.L of 5 Xprotein loading buffer, 10. mu.L of PMSF) was added to each well, lysed on ice for 17min, the lysate was scraped off and transferred to an EP tube, cooked at 100 ℃ for 20min, and placed in a freezer at-20 ℃ for use. The extracted protein concentration is determined by using a BCA kit and then SDS-PAGE electrophoresis is carried out, electrophoresis gel is transferred to an NC membrane by a semidry method, the NC membrane is sealed by 5% milk powder at 4 ℃ overnight, washed for 3 times by 1 XTSST, incubated for 2h at 37 ℃ by using antibody diluent (1: 1000 dilution) of HSV-1/HSV-2 ICP27 and gB, washed for 3 times, and then incubated for 2h at 37 ℃ by using secondary antibody diluent (1: 5000 dilution). The color was developed using the alkaline phosphatase test kit, and the Image was photographed and processed with Image J.

1.3 real-time fluorescent quantitative PCR (RT-PCR)

Inoculating Vero cells into a 6-well plate, infecting the cells for 1h at 37 ℃ by using HSV (MOI = 0.1) when the cells grow to a monolayer, then adding maintenance liquid containing wedelolactone with different concentrations into corresponding wells, and simultaneously arranging a virus control group and a blank control group. After 16h, total RNA was extracted using RNA extraction kit. After concentration detection of the extracted RNA, the RNA was subjected to One Step TB Green by using an Applied Biosystems 7500 Fast Real Time PCR System^® PrimeScript^TMThe RT-PCR Kit II Kit instructions were used for the procedure.

The amplification primer sequences are as follows:

HSV-1 (gD) mRNA Forward Primer：5’-AGCAGGGGTTAGGGAGTTG -3’；

HSV-1 (gD) mRNA Reverse Primer：5’-CCATCTTGAGAGAGGCATC -3’；

Actin mRNA Forward Primer：5’- CTCCATCCTGGCCTCGCTGT -3’；

Actin mRNA Reverse Primer：5’-GCTGTCACCTTCACCGTTCC -3’。

1.4 statistical analysis

All data are representative of at least three independent experiments. Data are presented as mean ± Standard Deviation (SD). Statistical significance was tested in GraphPad Prism 7 software using one way-ANOVA.P <0.05 was considered statistically significant.

2. Results of the experiment

2.1 inhibition of HSV by wedelolactone in different periods of time after adsorption

The experiments show that wedelolactone can inhibit HSV infection in the post-adsorption stage, so that the inhibition activity of wedelolactone in drug administration at different time stages after HSV infection is evaluated by using a Western blot test. As shown in figure 4, the wedelolactone can be very obviously reduced in virus titer after being adsorbed by the wedelolactone after being administered for 2-4h, and the inhibition effect on virus proliferation is also very obvious after 4-6 h. Almost has no inhibition effect for 6-8h, 8-10h and 10-12 h. In addition, the wedelolactone treatment at 0-6h or 0-12h has remarkable effect in inhibiting virus proliferation. Thus wedelolactone works best in the early stages, i.e. 2-4h, after HSV infection.

2.2 evaluation of Wedelia chinensis lactone inhibition effect on viral protein expression by Western blot test

The influence of wedelolactone on HSV virus protein expression is detected by adopting a Western blot test, and ICP27 protein of HSV-1 and gB protein of HSV-2 are selected for detection and quantitative analysis is carried out. The results of the experiment are shown in FIGS. 5 and 6. The result shows that the ICP27 protein of HSV-1 can hardly detect or only detect a very shallow protein band when the wedelolactone concentration is 5-10 mu M. Meanwhile, after gray level analysis and quantification, the wedelolactone has an obvious inhibiting effect on the expression of HSV-1 ICP27 protein at 1.25-2.5 mu M, and the expression quantity of HSV-1 ICP27 protein is reduced by 40-50%; the inhibition effect on the ICP27 protein expression is very obvious when the molecular weight is 5-10 mu M, and the expression quantity of ICP27 protein of HSV-1 is reduced by 80-90%.

In addition, the results in FIG. 6 show that the gB protein band of HSV-2 is hardly detected at drug concentrations of 1-2. mu.M. After gray scale analysis and quantification, the wedelolactone has a remarkable inhibiting effect on the expression of HSV-2 gB protein at 0.25 mu M; when the protein is 0.5-2 mu M, the inhibition effect on the expression of HSV-2 gB protein is very obvious, and the expression quantity of HSV-2 gB protein is reduced by 80-90%. The results show that wedelolactone can obviously reduce the expression level of HSV virus protein, and the inhibition effect has obvious dose dependence.

2.3 Wedelia lactone inhibition of HSV gD protein mRNA expression

Treating with wedelolactone, infecting Vero cell with HSV-1, extracting RNA 16 hr, detecting mRNA expression level of virus gD protein by RT-PCR method, and detecting with 2^-ΔΔCTMethod for processing experimental knotThe results are shown in FIG. 7. After infection by HSV-1, wedelolactone has obvious inhibition effect on the expression of mRNA of HSV-1 virus protein gD when the medicine concentration is 1.25 mu M, and has extremely obvious inhibition effect when the medicine concentration is more than 2.5 mu M. The experimental result is basically consistent with the detection result of the viral protein, and the experimental result and the detection result of the viral protein all present obvious dose dependence, which indicates that the wedelolactone can inhibit the replication process after the absorption of the virus, and the inhibition of the virus entry and the early replication process are possibly related.

In conclusion, the results show that wedelolactone has effective inhibitory activity against HSV-1 and HSV-2, and has very low toxicity. Wedelolactone can block HSV infection through direct action with virus particles, thereby interfering with virus adsorption and early replication processes. The action mechanism of wedelolactone is different from that of the existing HSV-resistant nucleoside drugs. Therefore, wedelolactone, a natural product from a plant source, is worthy of further study as a novel anti-HSV preparation in the future.

Example 3: wedelolactone for treating HSV-infected mice

1. Experimental methods

1.1 Experimental modeling

All the mice used in the invention meet the humanistic treatment guidance of experimental animals of the department of scientific technology of China (vgkfcz-2006-398). BALB/c female mice of 3-4 weeks old were used, weighing 14-15 g, and were randomly divided into 5 groups: a blank control group, a virus control group, a positive drug (acyclovir 10 mg/kg/day) group, a wedelolactone high-dose (5 mg/kg/day) group and a wedelolactone low-dose (2.5 mg/kg/day) group. The nasal drop method is adopted for receiving the toxin, and the drug administration is carried out 4 hours after the toxin receiving, and the drug administration is continuously carried out for 5 days.

1.2 mouse detection

And (3) dissecting 72 h after inoculation, taking the lung and spinal cord of the mouse, and verifying the effect of the medicine in resisting HSV-1 in vivo through a plaque test and real-time fluorescent quantitative RT-PCR. The virus inoculation day is taken as the initial day, the clinical symptoms of the mice are observed for 14 consecutive days, and the weight change and the survival condition of the mice are recorded every day.

2. Results of the experiment

2.1 weight Change in mice following HSV-1 infection

FIG. 8 is a diagram of a mouseWeight change. The results show that the blank group weight slightly decreases in the first 3 days, possibly related to environmental factors, but gradually increases overall. The viral group weight continued to decline for the first 6 days and began to rise slowly with slight fluctuations but still lighter than the original body weight after day 6 compared to the blank group. Positive drug group (10 mg. kg)^-1) The weight of the product slowly decreases in the first 3 days, and then slowly rises; wedelolactone processing group (5 mg kg)^-1Or 2.5 mg/kg^-1) The weight decreased slightly in the first 3 days, then began to rise slowly, although slightly fluctuating, but still slightly above the weight of the positive drug group.

2.2 survival Rate profiles in mice

FIG. 9 shows the survival rate of mice. The results show that the virus group began to die at day 2 after virus inoculation, and the survival rate decreased to 10% at day 13; positive drug group (10 mg. kg)^-1) Death begins to occur on day 3 after virus inoculation, and the survival rate is reduced to 60% on day 9; wedelolactone high dose group (5 mg kg)^-1) Death starts to occur on the 5 th day after virus inoculation, and the survival rate is reduced to 80% on the 7 th day; wedellactone low dose group (2.5 mg kg)^-1) Death began to occur on day 4 after inoculation and survival decreased to 60% on day 10. Compared with the virome, the survival rate (80%) of the wedelolactone high-dose group is obviously improved and is superior to that of the positive medicine group (60%). The wedelolactone can obviously improve the survival condition of the mice infected with HSV-1 virus and improve the survival rate of the mice.

2.3 viral load changes in spinal cord tissue in mice

The virus load in mouse tissue is an important index for analyzing virus infection, latent infection can be established in nerve tissue by HSV, and the virus load in mouse spinal cord tissue is detected by RT-PCR. The results are shown in FIG. 10, which shows that the positive drug group (10 mg. kg) is higher than the viral group^-1) Reduced viral load by about 90%, wedelolactone high dose group (5 mg kg)^-1) The virus load is reduced by nearly 98 percent, and the effect is slightly better than that of a positive drug acyclovir group; wedelia lactone low dose group (2.5 mg kg)^-1) Also reduce by about80% viral load. The results show that wedelolactone can obviously reduce the virus load in mice infected with HSV on the in vivo animal level, and the effect of a high-dose group is better than that of acyclovir serving as a positive drug. In summary, wedelolactone also has a very good in vivo anti-HSV infection effect.

The above embodiments are only used for illustrating the technical solution of the present invention, and not for limiting the same; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.

Claims

1. The application of Werethrin in the preparation of medicines and health products for preventing and/or treating herpes virus infection.

2. application according to claim 1 is characterized in that: described herpes virus is herpes simplex virus, is specially HSV-1 virus and HSV-2 virus.

3. application according to claim 1, is characterized in that: the use concentration of described wecarolide is 0.1 μM～50 μM.

4. application according to claim 2 is characterized in that: the use concentration that described wecarolide effectively inhibits HSV-2 virus is 0.25 μM～2 μM.

5. application according to claim 2, it is characterised in that: the use concentration that described wecarolide effectively inhibits HSV-1 virus is 1.25 μM～10 μM.

6. application according to claim 1, is characterized in that: described medicine and health-care product are main active component with wecarolide and its pharmaceutically acceptable salt, stereoisomer, crystal or derivative, It is prepared together with pharmaceutical auxiliary ingredients.

7. application according to claim 1 is characterized in that: the use time of described medicine and health care product is within 2～6 h after herpes virus infection.

8. application according to claim 7, is characterized in that: in described medicine and health product, the dosage that contains main active component wemarolide is 2.5 mg/kg/d～5 mg/kg/d.

9. application according to claim 1 is characterized in that: described medicine and health-care product reach the effect of blocking herpes virus infection through the direct action of the Werethrin and herpes virus contained therein.

10. a medicine for preventing and/or treating herpes virus infection, it is characterized in that: described medicine is main active ingredient with wecarolide and its pharmaceutically acceptable salt, stereoisomer, crystal or derivative .