An antiviral pharmaceutical composition containing a therapeutic agent derived from Astraeus Asiaticus

Abstract

The present invention relates to an antiviral pharmaceutical composition comprising a polar extract of Astraeus asiaticus; and one or more pharmaceutically acceptable excipients selected from solvents, additives, buffers, preservatives, flavoring agents and stabilizers. The composition also comprises acyclovir. The present invention also relates to the use of compounds for treating diseases caused by enterovirus 71 (EV71), coxsackievirus A16 (CA V16), herpes simplex virus type 1 (HSV‑1) and / or herpes simplex virus type 2 (HSV‑2), wherein HSV‑2 strains are resistant to acyclovir, wherein the diseases include hand, foot and mouth disease (HFMD). The present invention also relates to the use of ergosterol for treating hand, foot and mouth disease.

Description

Technical Field

[0001] The present disclosure relates to a pharmaceutical composition having antiviral properties. The disclosed composition comprises at least one active therapeutic agent derived from Astraeus asiaticus or a derivative thereof. Alternatively, the disclosed composition may have one or more antiviral drugs or agents to synergistically improve the therapeutic effect achieved. Background Art

[0002] Enterovirus 71 (EV71) and coxsackievirus A16 (CA16) belong to the Picornaviridae family and are pathogens that cause multiple pandemic diseases worldwide[1]. In particular, both EV71 and CA16 are the main causes of hand, foot and mouth disease (HFMD) and herpangina[2]. In general, EV71 is an RNA-based virus that can cause central nervous system (CNS)-related complications in humans, including cerebellar ataxia, acute brainstem encephalitis, and poliomyelitis-like paralysis[3]. More importantly, EV71 primarily infects infants and children under 5 years of age[4], who are more susceptible to the complications caused than adults. As a result, infants and children infected with HFMD have a higher mortality rate. Common symptoms of HFMD are blisters on the hands, soles of the feet, and buttocks or maculopapular rashes on the skin of infected subjects[5]. HFMD is easily transmitted through the fecal-oral route or through contact with surfaces contaminated by oropharyngeal secretions[6]. Unfortunately, there is currently no effective clinical drug available to treat or significantly alleviate the symptoms of HFMD patients[7].

[0003] Astraeus is a member of the Diplocystaceae (Astraeaceae) family and is a fungus commonly found in tropical countries [8]. Due to the widespread traditional knowledge in the region, there has been a lot of research on the medicinal value of different Astraeus members, including A. sirindhorniae [9], A. asiaticus, and A. odoratus

[10] in Southeast Asia. Some early studies reported that the most abundant compounds in Astraeus are triterpenoids. In these studies, extracts of A. odoratus containing astraodorols such as astraodoric acid A and B were found to have great anti-cancer cell proliferation and anti-tuberculosis potential

[11] . Modifications of the chemical structure of astraodorol also showed anti-malarial activity in vero cell lines

[12] . In addition, antibacterial and antifungal properties of astraodorol have been observed

[13] . Nevertheless, the effects of Astraeus-derived compounds and / or extracts on viral activity remain unclear. Therefore, it is highly worthwhile to explore the potential use of extracts, crude or purified, of A. asiaticus, especially from different plant parts and any other derivatives obtained thereof, in neutralizing harmful viral activity in subjects suffering from enteroviral infections. Summary of the invention

[0004] One object of the present disclosure is to provide an antiviral composition comprising at least one active therapeutic agent of natural origin. The inventors of the present disclosure have found that the active therapeutic agent has no or little cytotoxicity to a subject treated with the disclosed composition.

[0005] A further object of the present disclosure is to provide an antiviral pharmaceutical composition that can effectively inhibit viral replication of EV 71, CA 16, herpes simplex virus type 1 (HSV-1) and / or herpes simplex virus type 2 (HSV-2) in a subject. Therefore, the disclosed composition can be used as a means of treating conditions associated with or caused by these viruses.

[0006] Another object of the present disclosure is to obtain a synergistic therapeutic effect through the disclosed composition by incorporating one or more other antiviral compounds.

[0007] In addition, another object of the present disclosure relates to the use of ergosterol or any derivative obtained thereof for treating hand, foot and mouth disease.

[0008] One aspect of the present disclosure relates to an antiviral pharmaceutical composition comprising a polar extract of A. asiaticus; and one or more pharmaceutically acceptable excipients selected from solvents, additives, buffers, preservatives, flavoring agents and stabilizers. Preferably, the composition is effective against viral replication in a host cell infected with a predetermined virus.

[0009] Further embodiments of the disclosed compositions also include acyclovir, which has been found to synergize with the polar extract of Astraeus asiaticus against herpes simplex virus.

[0010] For more embodiments, for the disclosed antiviral composition, the weight ratio of the extract of A. asiaticus to acyclovir is 1:1 to 4:1.

[0011] For more embodiments, the concentrations of the polar extract and acyclovir in the disclosed composition are 60 to 500 ug / ml and 0.10 to 100 ug / ml, respectively.

[0012] In several embodiments, the polar extract used in the disclosed compositions is obtained by heating A. asiaticus with water for a predetermined time, removing the A. asiaticus from the water, and concentrating the water to produce a polar extract.

[0013] For more embodiments, the predetermined virus is any one or a combination of enterovirus 71, coxsackievirus A16, herpes simplex virus type 1, herpes simplex virus type 2, acyclovir-resistant herpes simplex virus type 1, and acyclovir-resistant herpes simplex virus type 2.

[0014] In some embodiments, herpes simplex virus type 1 and herpes simplex virus type 2 refer to strains that are resistant to acyclovir when acyclovir is used alone to combat infection caused.

[0015] Another aspect of the present disclosure relates to an aqueous extract of A. asiaticus for use in treating hand, foot and mouth disease.

[0016] In addition, another aspect of the present disclosure relates to the use of ergosterol or any derivatives obtained thereof for treating hand, foot and mouth disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 is the HPLC chromatogram of the aqueous extract of A. asiaticus showing the peak corresponding to ergosterol;

[0018] Figure 2 It is a graph showing the cytotoxic effects of different concentrations of aqueous extracts of A. asiaticus on Vero cells;

[0019] Figure 3 is a graph showing the results of the efficiency of the aqueous extract of A. asiaticus in inhibiting the entry of HSV 1 and HSV 2 into Vero cells pretreated with different concentrations of the extract;

[0020] Figure 4 is a graph showing the results of the efficiency of aqueous extracts of A. asiaticus as virucide in killing and / or inhibiting HSV 1 and HSV 2 infection of Vero cells treated with different concentrations of the extracts;

[0021] Figure 5 is a graph showing the results of the efficiency of the aqueous extract of A. asiaticus in inhibiting infection of HSV 1 and HSV 2 on Vero cells treated with different concentrations of the extract;

[0022] Figure 6 is a graph showing the results of the efficiency of aqueous extracts of A. asiaticus in inhibiting viral replication of HSV 1 and HSV 2 in infected Vero cells treated with different concentrations of the extracts; and

[0023] Figure 7 is a graph illustrating the results and efficiency of aqueous extracts of A. asiaticus (H2O [μg / ml]) in inhibiting viral replication of enterovirus 71 and coxsackievirus A16 in Vero cells pretreated with the extracts. DETAILED DESCRIPTION

[0024] Hereinafter, the present invention will be described according to preferred embodiments and with reference to the attached description and drawings. However, it should be understood that the description with reference to the preferred embodiments and drawings of the present disclosure is only for the convenience of discussing the various disclosed embodiments, and it is conceivable that various modifications may be made by those skilled in the art in the case of departing from the scope of the attached claims.

[0025] Unless otherwise noted, the terms "include" and "comprising" and grammatical variations thereof as used herein are intended to represent "open" or "inclusive" language such that they include the listed elements but also permit the inclusion of additional, non-listed elements.

[0026] As used herein, the phrase "an embodiment" or "an embodiment" means in some embodiments but not necessarily in all embodiments.

[0027] As used herein, in the context of component concentrations, conditions, other measurements, and the like, the term "approximately" or "about" means + / - 5% of the stated value, or + / - 4% of the stated value, or + / - 3% of the stated value, or + / - 2% of the stated value, or + / - 1% of the stated value, or + / - 0.5% of the stated value, or + / - 0% of the stated value.

[0028] The terms "aqueous extract" and "water extract" are used interchangeably throughout the description provided below and refer to an extract obtained by contacting A. asiaticus with water, preferably deionized water, under one or more predetermined conditions, promoting the dissolution of polar or water-soluble compounds present in A. asiaticus in the water, separating the water and the dissolved compounds from the A. asiaticus, and removing and concentrating the separated water, unless otherwise specified. Preferably, the A. asiaticus and water used in the extraction process are prepared in a ratio of 1:1 to 1:6 (by w / v).

[0029] The term "antiviral" or "virus inhibition" or "antiviral replication" refers to interference with the replication of the viral life cycle, including preventing the virus from penetrating into cells and / or directly killing the virus. However, it should not be interpreted as having a direct effect on the virus, such as preventing cell receptors from binding to the virus, inhibiting gene expression during viral replication, etc. "Killing the virus" refers to a direct effect on the virus itself, resulting in viral inactivation. The antiviral properties of A. asiaticus extracts mentioned below may refer to the ability of the extract to interfere with viral replication, which is achieved specifically by inhibiting the binding between host cell receptors and viruses, or by inhibiting the penetration of host cells, or even inhibiting or killing the virus after replication.

[0030] As used herein, the term "synergistic effect" may refer to a combination of at least two therapeutic agents in a treatment or therapeutic process, wherein the combination of at least two therapeutic agents produces a therapeutic effect greater than the sum of each therapeutic agent alone, for example, when acyclovir is combined with an extract of A. asiaticus, the resulting therapeutic result is very effective against viral replication in a synergistic manner, particularly against the replication of drug-resistant strains.

[0031] According to one aspect of the present disclosure, an antiviral pharmaceutical composition is disclosed. In essence, the disclosed antiviral composition comprises a polar extract of A.asiaticus; and one or more pharmaceutically acceptable excipients selected from solvents, additives, buffers, preservatives, flavoring agents and stabilizers. The disclosed composition is effective against viral replication in a host cell infected with a predetermined virus. For many embodiments, the disclosed composition is effective against, but not limited to, enterovirus 71, coxsackievirus A16, herpes simplex virus type 1 and / or herpes simplex virus type 2.

[0032] Preferably, the polar extract is obtained by heating the cleaned A.asiaticus with water or an aqueous phase at a temperature of 95 to 100° C. After the heat treatment that causes the desired polar compounds to leave the A.asiaticus matrix, the water with the dissolved compounds is separated from the spent A.asiaticus by one or more filtration steps to remove any unwanted solid residues from the aqueous phase. The aqueous phase free of solid residues is then concentrated by further heating or vacuum drying to precipitate a polar extract for the preparation of an antiviral pharmaceutical composition. It will be appreciated by those skilled in the art that other polar solvents such as solvent alcohols, acetonitrile, etc. may also be used for extraction, by mixing and contacting A.asiaticus with these solvents, with or without heating to bring out and dissolve the desired polar compounds. The inventors of the present disclosure have found that ergosterol may be one of the available active therapeutic agents in the polar extract, giving the disclosed composition antiviral properties.

[0033] In addition, the type of excipient used with the A. asiaticus aqueous extract depends largely on the preparation to be prepared to deliver a given bioactive compound in the extract to obtain the desired therapeutic effect. For example, the disclosed composition can be in an emulsified formulation for oral administration, and similar emulsified formulations can contain one or more buffers and / or stabilizers to preserve the prepared emulsion, thereby promoting the absorption of the bioactive compound. Alternatively, the disclosed composition can be in the form of a cream, lotion, ointment, etc., for topical application to the skin, especially the feet and hands of the subject where the injury is caused.

[0034] According to a more preferred embodiment, the disclosed composition may also contain acyclovir. In particular, acyclovir is a known antiviral drug for treating symptoms caused by certain types of viruses. For example, it has been used to treat herpes zoster caused by herpes labialis associated with herpes simplex and herpes zoster. Typically, acyclovir is diphosphorylated and triphosphorylated by the kinases of the infected virus and host cells to inhibit the operation of HSV DNA polymerase and cause chain termination. Although acyclovir is an antiviral drug, it cannot cure the infection caused by the above-mentioned viruses. It only reduces the number of outbreaks in infected subjects, reduces the severity, and shortens the duration of each outbreak. Nevertheless, what is more worrying is that the number of cases reported about HSV strains resistant to acyclovir treatment has been increasing. Through the experiments conducted, the inventors of the present disclosure have noticed that in the presence of acyclovir, the antiviral efficiency against HSV, especially HSV 1 and HSV 2, has been improved. More importantly, the HSV 1 and HSV 2 mentioned in these embodiments can be strains that are resistant to the treatment or therapeutic effect of acyclovir. Therefore, the synergistic therapeutic effect produced by the combined use of the aqueous extract of A. asiaticus and acyclovir can prevent the replication of HSV 1 or HSV 2, which refers to strains that are resistant to acyclovir when acyclovir is used alone.

[0035] For more embodiments, the weight ratio of the extract of A.asiaticus to acyclovir is 1:1 to 4:1 to achieve a synergistic therapeutic effect. Preferably, the ratio can be adjusted according to the formulation adopted by the disclosed composition to provide therapeutic results. More importantly, in addition to acyclovir, other antiviral drugs can be incorporated into the disclosed composition to cover a better broad spectrum against different viruses.

[0036] According to further embodiments, the concentration of the polar or aqueous extract may be 60 to 500 ug / ml, and the desired results are obtained after the disclosed composition is administered to a subject infected by a predetermined virus. Likewise, the concentration or amount of acyclovir used in the disclosed composition is about 0.20 to 12.50 ug / ml.

[0037] Another aspect of the present disclosure relates to an aqueous extract of A.asiaticus for use in treating hand, foot and mouth disease (HFMD). Similar aspects of the present disclosure may include an aqueous extract of A.asiaticus for use in treating herpes simplex infections caused by HSV 1 and / or HSV 2. For further embodiments, HSV 1 and HSV 2 may be strains that are resistant to treatment with acyclovir. Notably, the treatments described herein should include alleviating symptoms experienced by subjects infected with enterovirus 71, coxsackievirus A16, HSV 1 or HSV 2. Treatment may include improving lesion healing, reducing the time to outbreaks caused by the virus that occur in a subject, and / or shortening the time to occurrence.

[0038] More aspects of the present disclosure relate to ergosterol for treating hand, foot and mouth disease or any derivatives thereof obtained. The ergosterol used in these embodiments can be obtained from one or more natural sources, such as A.asiaticus or any other organism capable of spontaneously synthesizing it. Alternatively, the ergosterol used can be produced by chemical synthesis using chemical reactions known in the art.

[0039] The following examples are provided to more clearly illustrate the present invention, but these examples are not intended to limit the scope of the present invention.

[0040] Example 1

[0041] 500 g of fresh fruiting bodies of the two mushrooms were washed, boiled in 1500 mL of deionized water (1 g fresh weight: 3 mL water ratio) for 15 min, and filtered through Whatman No. 1 filter paper. The filtrate was freeze-dried and cooled to -20°C for later use. The yields of A. asiaticus were 0.42% and 1.48%, respectively.

[0042] Example 2

[0043] Vero cell lines were cultured in a 96-well plate in a CO2 incubator at 37°C. After overnight, the cultured cells were further incubated with different concentrations of A.asiaticus aqueous extract for 24 h, while DMSO was used as a control group. MTT solution was then added to the cells and incubated with the extract for another 4 h, and then DMSO was used to dissolve the formazan crystals and the OD values ​​were calculated. 560 Based on the results obtained, the experiments showed that both A. asiaticus aqueous extracts were able to inhibit the proliferation of Vero cells in the dose range of 7.80 μg / ml to 2000 μg / ml, and even at doses as high as 2000 μg / ml, this did not seem to produce detectable toxic effects on the Vero cell line ( Figure 2 ).

[0044] Example 3

[0045] Vero cells were cultured in Earle's essential medium (EMEM) containing 10% fetal bovine serum (FBS), penicillin and streptomycin using 24-well culture plates at 37°C and 5% CO2 for 24 hours. The culture medium in each well was then replaced and incubated at 37°C for 1 hour with fresh culture medium spiked with different concentrations of extracts at 62.5 μg / ml, 125 μg / ml, 250 μg / ml and 500 μg / ml. Again, the culture medium combined with the extract was replaced with fresh culture medium, and then the cells were incubated at 37°C with cells isolated from patient samples with herpes simplex virus type 1 (HSV 1) and herpes simplex virus type 2 (HSV 2) for 1 hour. Subsequently, the unattached virus was removed and the Vero cells were washed. The washed Vero cells were placed in a 1.2% microcrystalline cellulose containing The extracts were incubated in a 5% CO2 medium for 3 days at 37°C and 5% CO2. The manner in which the extracts prevented herpes simplex virus attachment was investigated using a plaque reduction assay.

[0046] It was found that the extract of A. asiaticus did prevent the attachment of herpes simplex virus type 1 and herpes simplex virus type 2 to the tested Vero cells with an efficiency of 50 to 100%, which corresponds to an extract concentration of less than or equal to 62.50 μg / ml to 500 μg / ml, as shown in Figure 3 The present disclosure has demonstrated that polar or aqueous extracts of A. asiaticus can prevent herpes simplex virus from attaching to host cells, thereby preventing the virus from entering and infecting cells by attachment.

[0047] Example 4

[0048] Vero cells were cultured and seeded in 24-well tissue culture plates in EMEM containing FBS, penicillin and streptomycin at 37°C and 5% carbon dioxide for 1 day. The culture medium was removed, and the Vero cells were then washed with cold PBS and contacted with different solvent mixtures containing extracts incubated with HSV 1 or HSV 2, respectively. In particular, the solvent mixtures were prepared by mixing aqueous extracts of A. asiaticus at concentrations of 62.5 μg / ml, 125 μg / ml, 250 μg / ml and 500 μg / ml with HSV 1 and HSV 2 isolated from patients, respectively, and incubating at 4°C for 1 hour, and then contacting them with Vero cells. Vero cells and the solvent mixture were incubated at 4°C for 1 hour. Subsequently, unattached viruses were removed and the Vero cells were washed with cold phosphate-buffered saline (PBS). The washed Vero cells were placed in another medium containing 1.2% microcrystalline cellulose The cells were further cultured at 37°C and 5% CO2 for 3 days, and the effect of the water extract was investigated using a plaque reduction assay.

[0049] The present disclosure found that when used at concentrations of 62.50 μg / ml to 500 μg / ml, aqueous extracts of A. asiaticus were able to kill 50-100% of HSV 1 and HSV 2 isolated from patient samples in all samples, such as Figure 4 Therefore, the present disclosure concludes that the aqueous extract of A. asiaticus is able to kill enveloped viruses HSV 1 and HSV 2 to prevent potential infections caused by these viruses.

[0050] Example 5

[0051] Vero cells were cultured and seeded in 24-well tissue culture plates in EMEM containing FBS, penicillin and streptomycin at 37°C and 5% carbon dioxide for 1 day. The culture medium was then removed and the Vero cells were washed with cold PBS. HSV 1 and HSV 2 isolated from patient samples were then added to the cultured cells and incubated at 4°C for 1 hour. Unattached viruses were removed and the Vero cells were washed with cold PBS solution. A. asiaticus aqueous extract at concentrations of 200 μg / ml, 300 μg / ml, 400 μg / ml and 500 μg / ml was added to the Vero cells and incubated at 4°C for 1 hour. The culture medium was then removed again and the Vero cells were washed with cold PBS solution. Finally, the washed Vero cells were placed in another medium containing 1.2% microcrystalline cellulose. The cells were incubated in fresh medium at 37°C and 5% CO2 for 3 days and studied using the plaque reduction assay.

[0052] Based on the results obtained, the present invention found that the aqueous extract of A. asiaticus was able to inhibit the entry of HSV 1 and HSV 2 into host cells with an efficiency of about 50 to 100%, which corresponds to a concentration range of 62.50 μg / ml to 500 μg / ml, as Figure 5 shown.

[0053] Example 6

[0054] Vero cells were cultured and seeded in 24-well tissue culture plates in EMEM containing FBS, penicillin, and streptomycin at 37°C and 5% carbon dioxide for 1 day. After removing the culture medium, the Vero cells were washed with PBS solution. HSV 1, HSV 2, enterovirus 71, and coxsackievirus A16 isolated from patient samples were contacted with the cells and then incubated at 37°C for 1 hour. Unattached viruses were removed and the Vero cells were washed with PBS solution. Finally, the Vero cells were placed in 1.2% microcrystalline cellulose containing The aqueous extract of A. asiaticus was added to fresh medium at a concentration of 200 μg / ml, 300 μg / ml, 400 μg / ml and 500 μg / ml and incubated at 37° C. for 3 days and 5 days, respectively. The efficiency of the aqueous extract in preventing viral replication in infected cells was investigated using a plaque reduction assay.

[0055] In view of the results obtained, the present disclosure found that the aqueous extract of A. asiaticus was able to inhibit the viral replication of HSV 1 and HSV 2 isolated from patient samples with an efficiency of about 50 to 100%, which corresponds to a concentration range of 400 μg / ml to 500 μg / ml of the aqueous extract of A. asiaticus, as Figure 6 In addition, the aqueous extract of A. asiaticus exhibited the ability to inhibit the viral replication of enterovirus 71 and coxsackievirus A16 with an efficiency of 50% to 100%, which corresponds to a concentration range of 100 μg / ml to 400 μg / ml, as shown in FIG. Figure 7 shown.

[0056] Example 7

[0057] Vero cells were cultured and seeded in 24-well tissue culture plates in EMEM containing FBS, penicillin and streptomycin at 37°C and 5% carbon dioxide for 1 day. The culture medium was then removed and the cultured cells were washed with PBS solution. Subsequently, the washed cells were contacted with HSV 1 and HSV 2 isolated from patient samples for further incubation at 37°C for 1 hour. Unattached viruses were removed and the Vero cells were then washed with PBS solution. The washed Vero cells were then placed in different solution groups: (a) extract group, (b) acyclovir group and (c) extract and acyclovir group. For the relevant groups, the water extract concentrations used were 500μg / ml, 400μg / ml, 300μg / ml, 200μg / ml and 100μg / ml, while the acyclovir concentrations used were 0.20μg / ml to 12.5μg / ml. The cultured cells, together with the corresponding solutions, were placed in a 1.2% microcrystalline cellulose containing solution. The extracts were then incubated in a culture medium of 5% CO2 and incubated for 3 days at 37°C and 5% CO2. The efficiency of the extracts and in combination with acyclovir in preventing HSV 1 and HSV 2 replication was then investigated using a plaque reduction assay.

[0058] According to the results obtained, the present disclosure found that the aqueous extract of A. asiaticus and acyclovir can act synergistically to inhibit the viral replication of HSV1 and HSV 2 in infected cells. In particular, the combination index value (CI value) was less than 1, indicating the synergistic effect of both the aqueous extract and acyclovir as shown in Table 1. The results further showed that HSV 1 and HSV 2 were resistant to treatment with acyclovir alone in all samples tested.

[0059] Table 1 shows the synergistic results of water-extracted A. asiaticus extract and acyclovir against herpes simplex virus from patient samples isolated from Thailand.

[0060]

[0061] Note: Combination Index (CI) <1 = synergistic effect, 1 = additive effect, 1> antagonistic effect

[0062] It should be understood that the present invention can be embodied in other specific forms and is not limited to the sole embodiment described above. However, modifications and equivalents of the disclosed concepts such as those readily apparent to those skilled in the art are intended to be included within the scope of the appended claims.

[0063] References

[0064] 1.Wang H, Li K, Ma L, Wu S, Hu J, Yan H, et al. Berberine inhibitsenterovirus 71 replication by downregulating the MEK / ERK signaling pathway and autophagy. Virol J. 2017; 14(1):2.

[0065] 2.Lee JT, Yen TY, Shih WL, Lu CY, Liu DP, Huang YC, et al. Enterovirus 71seroepidemiology in Taiwan in 2017 and comparison of those rates in 1997,1999and 2007.PLoS One.2019;14(10):e0224110.

[0066] 3.Chong P,Liu CC,Chow YH,Chou AH,Klein M.Review of enterovirus 71vaccines.Clin Infect Dis.2015;60(5):797-803.

[0067] 4.Yi EJ,Shin YJ,Kim JH,Kim TG,Chang SY.Enterovirus 71 infection andvaccines. Clin Exp Vaccine Res.2017;6(1):4-14.

[0068] 5.Lee MS,Chiang PS,Luo ST,Huang ML,Liou GY,Tsao KC,et al.Incidencerates of enterovirus 71 infections in young children during a nationwideepidemic in Taiwan,2008-09.PLoS Negl Trop Dis.2012;6(2):e1476.

[0069] 6.Chang LY,Tsao KC,Hsia SH,Shih SR,Huang CG,Chan WK,etal.Transmission and clinical features of enterovirus 71 infections inhousehold contacts in Taiwan.JAMA.2004;291(2):222-7.

[0070] 7.Wang SM,Liu CC.Enterovirus 71:epidemiology,pathogenesis andmanagement. Expert Rev Anti Infect Ther.2009;7(6):735-42.

[0071] New Phytol.2012:194(4):1079-95.8.Wilson AW,Binder M,Hibbett DS.

[0072] 9.Phosri C,Watling R,Suwannasai N,Wilson A,Martin MP.A newrepresentative of star-shaped fungi:Astraeus sirindhorniae sp.nov.fromThailand.PLoS One.2014;9(5):e71160.

[0073] [ PubMed ] 10.Phosri C,Martin MP,Sihanonth P,Whalley AJ,Watling R.Molecular studies of the genus Astraeus.Mycol Res.2007:111(Pt 3):275-86.

[0074] 11.Arpha K,Phosri C,Suwannasai N,Mongkolthanaruk W,SodngamS.Astraodoric acids AD:new lanostane triterpenes from edible mushroomAstraeus odoratus and their anti-Mycobacterium tuberculosis H37Ra andcytotoxic activity.J Agric Food Chem.201260(39):9834–41.

[0075] 12.Nasomjai P,Arpha K,Sodngam S,Brandt SD.Potential antimalarialderivatives from astraodorol.Arch Pharm Res.2014:37(12):1538-45.

[0076] 13.Kope HH,Tsantrizos YS,Fortin JA,Ogilvie KK.p-Hydroxybenzoylformicacid and(R)-(-)-p-hydroxymandelic acid,two antifungal compounds isolated fromthe liquid culture of the ectomycorrhizal fungus Pisolithus arhizus.Can JMicrobiol. 1991;37(4):258-64.

Claims

1. Use of the aqueous extract of Astraeus asiaticus in the preparation of antiviral drugs, characterized in that: The water extract of Astraeus asiaticus is obtained by heating Astraeus asiaticus and water, filtering and concentrating; the virus antagonized by the antiviral drug is enterovirus 71 or coxsackievirus A16; the concentration of the water extract of Astraeus asiaticus is 100 μg / mL-400 μg / mL.

2. Use of an aqueous extract of Astraeus asiaticus in the preparation of an antiviral drug, characterized in that the aqueous extract of Astraeus asiaticus is obtained by heating Astraeus asiaticus and water, filtering, and concentrating; the virus antagonized by the antiviral drug is herpes simplex virus type 1 or herpes simplex virus type 2; the concentration of the aqueous extract of Astraeus asiaticus is 62.5 μg / mL-500 μg / mL.

3. A composition of antiviral drugs, characterized in that: The invention is composed of a water extract of Astraeus asiaticus and acyclovir; the water extract of Astraeus asiaticus is obtained by heating Astraeus asiaticus and water, filtering and concentrating the water extract; the viruses antagonized by the antiviral drug composition are herpes simplex virus type 1 and herpes simplex virus type 2; in terms of μg / mL, the concentration ratio of the water extract of Astraeus asiaticus to the acyclovir is 100:0.2, 200:0.2, 300:0.2, 400:0.2 or 500:0.

2.

4. A composition for use in the preparation of an antiviral drug, characterized in that: The composition consists of an aqueous extract of Astraeus asiaticus and acyclovir; the aqueous extract of Astraeus asiaticus is obtained by heating Astraeus asiaticus and water, filtering, and concentrating; the viruses antagonized by the antiviral drug are herpes simplex virus type 1 and herpes simplex virus type 2; and the concentration ratio of the aqueous extract of Astraeus asiaticus to the acyclovir is 100:0.2, 200:0.2, 300:0.2, 400:0.2, or 500:0.2 in μg / mL.

5. An antiviral drug composition, characterized in that: The composition comprises a water extract of Astraeus asiaticus and acyclovir; the water extract of Astraeus asiaticus is obtained by heating Astraeus asiaticus and water, filtering, and concentrating; the virus antagonized by the antiviral drug composition is herpes simplex virus type 2 resistant to acyclovir; in terms of μg / mL, the concentration ratio of the water extract of Astraeus asiaticus to the acyclovir is 200:0.2, 300:0.2, 400:0.2, or 500:0.

2.

6. Use of a composition in the preparation of an antiviral drug, characterized in that: The composition consists of an aqueous extract of Astraeus asiaticus and acyclovir; the aqueous extract of Astraeus asiaticus is obtained by heating Astraeus asiaticus and water, filtering, and concentrating; the virus antagonized by the composition of antiviral drugs is herpes simplex virus type 2 resistant to acyclovir; in terms of μg / mL, the concentration ratio of the aqueous extract of Astraeus asiaticus to the acyclovir is 200:0.2, 300:0.2, 400:0.2, or 500:0.2.

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