KR20130095010A - Natural anti-herpes virus agent and compositions comprising thereof - Google Patents

Abstract

PURPOSE: A natural anti-herpes virus agent is provided to have excellent anti-herpes virus activity and to produce pharmaceutical, cosmetic, and food compositions with safety and efficacy. CONSTITUTION: An anti-herpes virus agent contains one or more extracts prepared from ingredients selected among Agrimonia pilosa L., Allium thunbergii G. Don, Poria cocos, Acer tegmentosum Maxim., Paeonia suffruticosa Andr., Scutellariae radix, or Artemisia annua L. as active ingredients. A pharmaceutical composition for preventing and treating herpes virus infectious diseases contains the anti-herpes virus agent as an active ingredient. A cosmetic composition for preventing and treating herpes virus infectious diseases contains the anti-herpes virus agent as an active ingredient. [Reference numerals] (AA) Experimental example of medicinal herb on a lesion; (BB) Control cell; (CC) Cell+virus+medicinal herb ingredient; (DD) Cell+virus; (EE) Cell+virus+tamiflu; (FF) Cell+virus+acyclovir; (GG) Cell+virus+echinacea

Description

Natural anti-herpesvirus agents and compositions comprising them [0002]

The present invention relates to a natural antifoaginous virus agent exhibiting an inhibitory effect on the growth of herpes zoster infection including herbal plant extracts, and a pharmaceutical, cosmetic and food composition containing the same.

Human herpesviruses are known to have eight types. They are latent in the body after infection, reactivated frequently according to their health status, and then developed or recovered. The biological properties of host cells and infected cells are classified into subfamilies of alpha, beta and gamma. Especially alpha aphasia is a herpes simplex virus which is latently infected with mucosa, epithelial cells and ganglion of human skin. (HSV-1), type 2 (Herpes simplex virus type 2, HSV-2) and varicella-zoster virus (VZV) , Is a viral disease that requires treatment and prevention accompanied by severe pain. Among them, herpes simplex virus 1 also causes dental infection, stomatitis, lips herpes, cornea and herpes encephalitis, and herpes simplex virus type 2 is also classified as genital herpesvirus. Shingles virus causes chickenpox at the time of initial infection and causes shingles when reactivated. BETA and GammaAsia have a limited range of symptoms compared to the alpha-amyloidosis virus. Cetomegalovirus (CMV), human herpesvirus type 6 or type 7 (human herpes type 6 or type 7, HHV-6 or HHV-7). CMV is infected by immune immature fetuses, immune-compromised AIDS patients, organ transplant recipients, cancer patients through inapparent infection at the time of initial infection, and manifests as host CMV encephalitis, CMV retinitis, and pneumonia. HHV-6 and HHV-7 cause sudden onset of infants. Epstein-Barr virus (EBV) and human herpes type 8 (HHV-8) are known to be present in Gamma asiatica. EBV is infectious mononucleosis, Causing tumors and causing B-lymphoma in patients with AIDS or organ transplants. HHV-8 is known to cause Kaposi's sarcoma and B-cell lymphoma in AIDS patients.

Conventionally developed antiviral drugs include acyclovir, amatadine, rimantadine, and zanamivir, and are synthetic drugs for external or oral drugs that inhibit the DNA synthesis of target viruses. Frequent use of these antiviral agents is reported to have side effects such as hypersensitivity, mental neuropathy, digestive system symptoms, and autonomic nervous system symptoms. Therefore, a lot of caution is needed and inevitably leads to resistance to medicines and cross resistance between medicines. The herpes virus vaccine belonging to the Alpha subspecies has not been commercialized yet, and the preventive measures are still insufficient.

For this reason, the traditional herbal medicine herbal medicine ingredients are extracted, and the anti-herpes virus product with high efficacy that can be routinely used due to the development of natural antiseptic virus material and product which does not cause any problem in stability and resistance Development is demanded.

Korean Patent No. 10-0347651

The present invention is to provide an antiseptic virus agent using a natural product extract showing a high inhibitory effect against herpes zoster virus infection.

The present invention provides an antiseptic agent comprising as an active ingredient, any one or more extracts selected from the group consisting of Seongeuksu, Mountain Leek, Baekbokryeong, Sangcheok, Mokdepi, Golden or Dogwood.

The present invention also provides a pharmaceutical composition for preventing and treating herpes zoster infectious diseases comprising the anti-herpesvirus agent as an active ingredient.

The present invention also provides a cosmetic composition for preventing and treating herpes zoster infectious diseases comprising the anti-herpesvirus agent as an active ingredient.

The present invention also provides a food composition for preventing and treating herpes zoster virus infection comprising the anti-herpesvirus agent as an active ingredient.

The present invention relates to a natural antifojin virus agent comprising an herbal extract as an active ingredient and having excellent antiposonic virus activity, and can provide medicines, cosmetics and food compositions having safe and effective efficacy by using the anti-herpes virus agent according to the present invention have.

1 shows a monolayer cultured cell of an antiviral experimental host cell.
Figure 2 shows cytotoxicity experiments by treatment with herb extracts on monolayer cultured cells.
Fig. 3 shows results of herpes zoster virus infection (virus attack experiment) on monolayer cultured cells.
Fig. 4 shows the results of an antiseptic virus activity test (CPE experiment) of herbal extract components.
Fig. 5 shows the results of an efficacy test (MTT test) of herpes simplex virus type 1 herbal extract components.
6 shows the results of an efficacy test (MTT test) of the herpes simplex virus type 2 herbal extract component.
7 shows the efficacy test (MTT experiment) of herpes zoster virus against herbal extract components.
Fig. 8 shows the results of LC-MASS instrumental analysis of the components of the extract of Zenchigaku (strawberry) herb.
FIG. 9 shows the results of LC-MASS instrumental analysis of the extract components of the acid leek extract.
Fig. 10 shows the result of LC-MASS instrumental analysis of the extract of the saponin extract.
Fig. 11 shows the result of LC-MASS instrumental analysis of extract of Baekbokryeong.

The present invention relates to Agrimonia pilosa LEDEB.), mountain leek ( Allium thunbergii G. Don, Poria cocas Wolf, Acer tegmentosum Maxim., Paeonia suffruticosa ), golden ( Scutellaria baicalensis Georgi) or Artemisia annua Linne) as an active ingredient.

The anti-herpesvirus agent of the present invention may be an extracting component of herbal medicine and may include glycosides such as glycolipid and glycoprotein, and saponins, flavonoids, fragrance components, other pigment components, and cell components.

The herbal extract may be a freeze-dried antiviral agent prepared by concentrating a hot-water extract or an ethanol extract in a vacuum or a composition containing the anti-apocclusive agent. The anti-herpes virus agent in the present invention means an agent which weakens or extinguishes the action of the virus invaded into the human body as an antiviral agent.

 It is obvious to a person skilled in the art that antivenin antiviral agents do not normally contain pathogenic microorganisms and thus are harmless to the human body even if they are ingested or used as external preparations.

The herpes zoster virus infection, which is an anti-herpes target, may be an infection by herpes simplex virus type 1, herpes simplex virus type 2 or herpes zoster virus.

The present invention also provides a pharmaceutical composition for preventing and treating herpes zoster infectious diseases comprising an anti-herpesvirus agent according to the present invention as an active ingredient.

The medicinal composition may be formulated into an appropriate carrier, an excipient, a disintegrant, a sweetener, a coating agent, a swelling agent, a lubricant, a lubricant, a flavoring agent, an antioxidant, a buffer, a bacteriostatic agent, a diluent, A lubricant, a binder, and a lubricant.

Specific examples of carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulite

The solid preparations for oral administration may be in the form of tablets, pills, powders, granules, capsules, powders, granules, powders, granules, These solid preparations can be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. For oral use

Examples of the liquid preparation include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to commonly used simple diluents such as water and liquid paraffin. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil,

Injectable esters such as ethyloleate and the like can be used. As the suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

The pharmaceutical composition may be selected from the group consisting of granules, powders, coated tablets, tablets, pills, capsules, suppositories, gels, syrups, juices, suspensions, emulsions, drops or solutions.

The pharmaceutical composition of the present invention may be administered orally or parenterally through intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, percutaneous, intranasal, inhalation, topical, rectal, 0.0 > a < / RTI >

The dosage of the pharmaceutical composition for preventing and treating herpes zoster infectious disease comprising the herb extract of the present invention as an active ingredient may be varied depending on the condition and body weight of the subject, the type and degree of the disease, the drug form, And may be suitably selected by those skilled in the art. The dosage of the pharmaceutical composition for preventing and treating herpes zoster infectious disease comprising the herb extract as an active ingredient is not particularly limited, but the daily dose may be 0.0001 to 100 mg / kg, 0.001 to 100 mg / kg, or 0.01 to 100 mg / mg / kg. The administration may be performed once a day or divided into several times, and thus the scope of the present invention is not limited thereto.

In the present invention, the "subject" may be a mammal including a human, but is not limited to these examples.

The present invention also provides a cosmetic composition for preventing and treating herpes zoster infectious diseases comprising the anti-herpesvirus agent according to the present invention as an active ingredient.

The cosmetic composition according to the present invention is a raw material or an article which is commonly used as an external preparation for skin to prevent herpes virus or alleviate or improve herpes zoster.

The cosmetic composition may further comprise at least one additive selected from the group consisting of diluent water, gel, dispersant, vitamin, surfactant, emulsifier, perfume, colorant, stabilizer, preservative, antioxidant, ultraviolet screening agent, pH adjuster and chelating agent As shown in FIG.

The cosmetic composition of the present invention for use in the prevention and treatment of herpes virus infection diseases is not limited to the formulation, but may be, for example, a softening agent, a nutritional lotion, a massage cream, a nutritional cream, a pack, a gel, Lipstick, makeup base, foundation, lotion, ointment, gel, cream, cleansing, cleanser, soap, shampoo, rinse, treatment and serum. Such cosmetics may contain conventional ingredients such as aqueous vitamins, oily vitamins, high molecular weight peptides, polymeric polysaccharides, sphingolipids and the like, and may be easily prepared according to techniques well known to those skilled in the art.

The cosmetic composition according to the present invention has no particular limitation on the other ingredients other than the natural antiseptic agent as an essential ingredient in the indicated proportions and may further contain ingredients for ordinary cosmetic preparation, but is not limited thereto. Functional cosmetics may also be included in the cosmetics.

The functional cosmetics are those to which an additional function is imparted to a cosmetic to function and express the function of the composition by a physical, biochemical, or biotechnological method. The cosmetics are used for controlling the body defense rhythm, And the like. The term " composition "

The cosmetic composition may be added to cosmetics added to or processed into raw materials of a spray, a skin lotion, a cosmetic, a cream, a cosmetic, a functional cosmetic, a patch, a cosmetic mask, a toilet paper containing antiviral component, And may be added with cosmetically acceptable auxiliary additives. In this case, the amount of antifoaginic agent added to the cosmetic composition may be 0.01 to 90% by weight, 0.1 to 90% by weight or 1 to 90% by weight of the total cosmetics, 0.0001 to 0.2 g / ml, 0.005 To 0.2 g / ml or 0.005 to 0.1 g / ml, but the present invention is not limited thereto.

The present invention also provides a food composition for preventing and treating herpes zoster infectious diseases comprising an anti-herpesvirus agent according to the present invention as an active ingredient.

The food composition according to the present invention can be generally used for foods commonly used in beverages, confectionery, health foods and the like containing an antiseptic virus agent. More specifically, but not exclusively, it is also possible to use other ingredients such as confectionery, sugars, ice cream products, dairy products, meat products, fish meat products, tofu or glutinous rice, edible oils, noodles, Dried ginseng products, kimchi pickles, dried foods, dried fruits, vegetables, fruits or vegetables, cutting products, fruit juice, vegetable juice, mixed juice, chips, noodles, processed livestock, It can be used for the production of foods such as fermented milk food, bean curd food, cereal food, fermented microorganism food, confectionery bakery, condiments, meat processing industry, acidic beverage, licorice and herb.

The food composition may further comprise at least one additive selected from the group consisting of organic acids, phosphates, antioxidants, lactose casein, dextrin, glucose, sugar and sorbitol.

The food composition may be in the form of a powder, a granule, a tablet, a capsule or a drink, although not limited thereto.

The food composition according to the present invention means a natural product or a processed product containing one or more nutrients and means that the food composition can be directly eaten through a certain processing step.

The food composition according to the present invention is not particularly limited to ingredients other than those containing the antivenzin virus agent as an essential ingredient in the indicated ratios and may contain a flavoring agent or a carbohydrate as an additional ingredient, But is not limited thereto. The food may also contain a functional food.

The above-mentioned functional food refers to a food group that imparts an additional function to function or express the function of the food by using physical, biochemical, biotechnological techniques, etc., or to control the bio-defense rhythm of the food composition, And the like. The term " food "

In another embodiment of the present invention, the food composition for the prevention and treatment of herpes zoster virus infection further comprises at least one additive selected from the group consisting of organic acids, phosphates, antioxidants, lactose casein, dextrin, glucose, sugar and sorbitol . The organic acid can be, but is not limited to, citric acid, fumaric acid, adipic acid, lactic acid or malic acid, and the phosphate can be sodium phosphate, potassium phosphate, acid pyrophosphate or polyphosphate (polymeric phosphate) But are not limited to, natural antioxidants such as polyphenols, catechins, alpha-tocopherol, rosemary extract, licorice extract, chitosan, tannic acid or phytic acid.

The food composition for preventing and treating herpes zoster virus infection of the present invention may contain various herb extracts, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, (Cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, . In addition, the food composition for preventing and treating herpes zoster virus infection according to one embodiment of the present invention may contain flesh for the production of natural fruit juice, fruit juice drink and vegetable drink. The proportion of such an additive is generally, but not limited to, 0 to 20 parts by weight, 5 to 20 parts by weight, or 5 to 15 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention can also be added with a food-acceptable food-aid additive. In this case, the amount of the antiseptic agent used in the food may be 0.01 to 90% by weight, 0.1 to 90% by weight or 1 to 90% by weight of the total food, 0.0001 to 0.2 g / ml, 0.005 to 0.2 g / ml, or 0.005 to 0.1 g / ml, but the present invention is not limited thereto.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example >

Example 1. Purification of an anti-herpes zoster virus extract component

Materials and Methods

(1) hot water extract

100 g of dried seaweed, bamboo shoots, bamboo shoots, sanggok mackerel, and white mugwort were added to 1000 ml of distilled water, boiled for 30 minutes, and centrifuged at 4000 rpm for 5 minutes. The supernatant was filtered with a 0.45 μm membrane filter and subdivided into a 15 ml sterile test tube It was used in the experiment while being kept frozen at -20 ℃. The herb was purchased from Kyungnam Agricultural Cooperative Federation.

(2) Ethanol extract

100 g of dried Seokhwakseong, Sungchulchu, Bongbyeol, Sanggolmyeong, and Dogum mugwort were placed in 1000 ml of ethanol (ethanol) for 24 hours at room temperature. Then, the alcohol extracts were filtered with a filter cloth, concentrated in vacuo, and lyophilized It was subdivided into 1 ml aseptic storage test tube and used in the experiment while being kept at -20 ℃ freezing.

Example 2. LC-MASS analysis of herbal extract ingredients

 (1) Experimental material

The extract was used for LC-MS analysis. The hot water extract was prepared by extracting 5.0 g dry weight of each material with 50 ml hot water and filtering the extract with a 0.2 μm membrane filter.

(2) reagents and appliances

All of the reagents and solvents used in the experiments were purchased from Agilent Technologies 6530 qTOF (Agilent Technologies, USA, Santa Clara, Calif.) Using LC-MS / MS. Instrumental analysis conditions are as follows, and MS ionization is defined as Positive ionization mode and Negative ionization mode.

LC Condition LC Condition device Agilent Technologies 6530 qTOF Culmn Halo C18 (2.1 mm * 150 mm, 2.7 Culumn temperature 40 ℃ Injection Volume 5 menstruum A: 0.1% Formic Acid in Water B: 0.1% Formic Acid in Acetonitrile Flow rate 0.6 ml / min

Solvent Gradient Condition of HPLC Time (min) B solution (%) 0 0 5 6 10 30 15 60 20 100 25 100

(3) LC-MS analysis results

Total ion chromatograms were obtained using LC-MS (q-TOF). The analytical data were analyzed using Agilent MassHunter Qualitative Analysis Software.

The chromatograms of positive ion mode and negative ion mode of LC-MS were examined for each sample, respectively.

LC-MS results of Seon Hakki Peak No. Name RT (min) Mass Formula Score Score (MFG) Height One n.n. 0.687 482.0639 C20 H14 N6 O7 S 48.3 96.6 6460 2 n.n 2.318 634.0738 C26 H22 N2 O15 S 45.74 91.47 6281 3 n.n 3.761 248.0285 C7 H8 N2 O8 46.78 93.56 40997 4 n.n 5.26 358.1218 C11 H22 N2 O11 46.19 92.38 9334 5 n.n 5.374 466.106 C16 H22 N2 O14 47.34 94.68 12015 6 n.n 5.765 466.1065 C16 H22 N2 O14 49.32 98.65 27966 7 n.n 6.623 434.0441 C14 H14 N2 O14 48.35 96.69 13016 8 n.n 6.691 466.1061 C16 H22 N2 O14 46.8 93.6 15447 9 n.n 6.746 450.1111 C13 H14 N12 O7 47.36 94.71 16156 10 n.n 6.825 302.0025 C9 H6 N2 O10 48.96 97.91 30521 11 n.n 7.405 462.0746 C16 H18 N2 O14 47.08 94.16 9100 12 n.n 8.335 446.0797 C16 H18 N2 O13 46.15 92.3 9828 13 n.n 12.489 476.1626 C16 H20 N12 O6 46.8 93.6 9252 14 n.n 12.489 314.1119 C14 H14 N6 O3 48.49 96.99 13143 15 n.n 13.938 254.112 C9 H14 N6 O3 46.42 92.84 8704

LC-MS results of sancheong Peak No. Name RT (min) Mass Formula Score Score (MFG) Height One Cellobiose 0.543 342.1149 C12 H22 O11 75.27 76.3 12806 2 Cordycepic acid 0.557 192.0628 C7 H12 O6 97.16 97.55 27436 3 Gallic acid 0.851 170.0208 C7 H6 O5 83.95 84.04 10987 4 Ergoflavine 0.993 610.1301 C30 H26 O14 67.64 67.79 5046 5 6-O-Galloyl-glucose 1.142 332.0743 C13 H16 O10 98.79 98.9 11823 6 3-Carboxy-4-hydroxy-phenoxy glucoside 1.273 316.0786 C13 H16 O9 80.07 80.95 10948 7 L-Epigallocatechin 1.275 306.0729 C15 H14 O7 93.47 93.66 19734 8 Biondnoid I 1.495 594.1353 C30 H26 O13 64 64.28 6263 9 Salidroside 2.507 300.1201 C14 H20 O7 95.32 95.59 47708 10 Apigenin-7-O-beta-D- (6'-p-hydroxy-cinnamoyloxy) -mannoside 2.592 578.14 C30 H26 O12 87.45 87.54 10919 11 Cuchiloside 2.9 432.1618 C19 H28 O11 89.42 90.01 15599 12 Epicatechin 3.046 290.0783 C15 H14 O6 95.26 95.93 47694 13 Marmesin 3.047 246.0882 C14 H14 O4 80.63 81.25 5795 14 2,3-Di-O-galloyl-D-glucose 3.282 484.0842 C20 H20 O14 96.88 96.89 11983 15 Cuchiloside 3.487 432.162 C19 H28 O11 94.54 94.51 26117 16 Fraxin 4.487 370.0893 C16 H18 O10 96.8 97.06 26636 17 1,2,3-Tri-O-galloyl-beta-D-glucose 5.252 636.0947 C27 H24 O18 93.8 94.36 11237 18 Curculigoside B 6.173 452.1328 C21 H24 O11 91.04 91.62 117987 19 Quercetin-3-L-arabino-7-D-glucoside 6.539 596.1368 C26 H28 O16 94.69 95.32 12813 20 Desmanthin 1 6.655 616.1056 C28 H24 O16 96.88 97.14 10096 21 Quercetin-3-L-arabino-7-D-glucoside 6.674 596.1375 C26 H28 O16 95.5 96.62 11313 22 (-) - Epicatechin-3-O-gallate 6.941 442.0898 C22 H18 O10 80.49 80.85 5813 23 8-Hydroxyluteolin-8-beta-D-glucopranoside 7.078 464.0953 C21 H20 O12 98.29 98.36 13127 24 Polystachoside 7.638 434.085 C20 H18 O11 98.89 98.94 7451

LC-MS results of mountain leek Peak No. Name RT Mass Formula Score Score (MFG) Height One gamma-L-Glutamyl-L-phenylalanine 3.027 294.1209 C14 H18 N2 O5 97.66 97.84 18231 2 Gentiatibetine 3.027 165.0783 C9 H11 N O2 81.9 82.68 5114 3 26-O-beta-D-Glucopyranosyl (25R) -5alpha-furostane-12-one-3beta, 22alpha, 26-triol-3-0beta-D-glucopyran 7.372 934.4776 C45 H74 O20 94.56 6386 4 Protoeruboside B 9.367 1260.599 C57 H96 O30 99.05 10943 5 Macrostemonoside L 10.276 918.4842 C45 H74 O19 92.24 18438 6 (25S) -26-O-beta-D-Glucopyranosyl-22-hydroxy-5beta-furostane-3beta, 26-diol 3-0beta-D-glucopyranosyl- 10.502 920.4985 C45 H76 O19 94.63 9747 7 Sanleng acid 12.805 330.2409 C18 H34 O5 98.55 98.59 12348 8 Terrestide B 12.986 754.4138 C39 H62 O14 93.19 8005

LC-MS results of Baekbok-ri Peak No. Name RT Mass Formula Score Mass (MFG) Score (MFG) Height One Harman 0.632 182.0824 C12 H10 N2 58.21 182.0844 59.42 14004 2 Afzelechin- (4alpha - > 8) -afzelechin 0.635 546.1522 C30 H26 O10 64.74 546.1526 65.41 11552 3 Dictangustinee 0.635 215.0602 C12H9NO3 56.98 215.0582 57.09 92280 4 Theobromine 0.639 180.0669 C7 H8 N4 O2 67 180.0647 67.09 25112 5 Aloeresin G 0.643 538.1823 C29 H30 O10 65.3 538.1839 66.08 10598 6 1-Formyl-beta-carboline 0.646 196.0628 C12 H8 N2 O 80.63 196.0637 80.68 385980 7 Carboline-1-carboxylic acid, methyl ester 0.647 226.073 C13 H10 N2 O2 67.64 226.0742 67.93 12697 8 6-Hydroxymethyllumazine 0.651 194.0463 C7 H6 N4 O3 64.76 194.044 65.42 29566 9 Ohioensine 0.661 372.0964 C23 H16 O5 51.69 372.0998 52.02 57375 10 Isopimpinellin 0.665 246.0551 C13 H10 O5 60.66 246.0528 62.49 13228 11 Flavidin 16.326 240.0782 C15 H12 O3 77.65 240.0786 78.83 15565

Example 3. Cytotoxicity test [

(1) viruses and cells

The herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) were distributed at the Konkuk University Medical School and the herpes zoster virus (VZV) Respectively.

Human fibroblasts (MRC-5) were purchased from the American Society of Microbial Communities and used for experiments. The viral strains were used by passaging on Vero cell lines cultured in DMEM medium (10% FBS, 1% (Penicillin 100 IU / ml, Streptomycin 100 IU / ml)).

(2) antiviral control substance

Echinacea, which is a drug that is effective against most herpesviruses, and Acyclovir, a drug used as an antiviral agent extracted from natural products, and Tamiflu were used as a control. Acyclovir and Echinacea were purchased from Sigma and dissolved in sterilized water at concentrations of 100 μg / ml and 36 μg / ml, respectively. Tamiflu was dissolved in water and then dissolved in water at a concentration of 750 μg / ml And used as a positive control.

(3) Cytotoxicity test

Vero cells and MRC-5 cells were inoculated into a T25 flask containing DMEM medium (containing 10% FBS and 1% penicillin streptomycin), cultured in a 5% CO 2 incubator at 37 ° C for 2-3 days, Vero cells were seeded at 4 x 103 cells / well and cultured for 2 days. The herbal extract extracts were treated with serum-free culture medium at a concentration of 2-fold to 70-80% of the formed monolayer cells, and cytotoxic effect (CPE) was observed. As a control, 0.89% physiological saline was used. After culturing, the cells were stained with a crystal biotite staining reagent, washed with running water, and observed with a microscope for cell observation. As a result, the cytotoxicity of the antiviral agent in the two-fold diluted sample of the herb extract was not significantly different from that of the control cells, and thus the safety of the herb extract used in the experiment was very high there was.

Rating of response rating

1) Grade (4): Inhibition of virus damage 95% or more Normal cell maintenance

2) Grade (3): Inhibition of virus damage 90% or more Normal cell maintenance

3) Grade (2): Inhibition of virus attack 50% or more Normal cell maintenance

4) Grade (1): Inhibition of virus attack 20% or more Normal cell maintenance

5) Grade (0): Inhibition of virus attack 5% or less Normal cell maintenance

Levels of cytotoxicity of host cells by herpes simplex virus type 1 and type 2 Plants name Target cell Score * Extracts Hot water Ethanol HSV-1 HSV-2 HSV-1 HSV-2 Agrimonia pilosa Ledebour)





Vero




4 4 4 4 Mountain leek ( Allium thunbergii G.Don) 4 4 3 2 Poria cocas Wolf) 3 4 4 2 Acer tegmentosum Maximowocz) 4 4 4 3 Paeonia suffruticosa ) 3 4 3 4 Golden ( Scutellaria baicalensis Georgi) 3 3 3 3 Artemisia annua Linne) 4 4 4 3 Mixture (Seokhogcho + Mountain Leek + Baekbongseong + Socheongmyeong + Dogwood Mugwort) 4 4 4 4 virus HSV-1 HSV-2 Acyclovir (RFID) 4 4 Echinacea 3 3 Tamiflu 4 4 Saline (Control) 0 0

Levels of cytotoxicity of host cells by shingles virus Plants name Target cell Score * Extracts Hot water Ethanol VZV VZV Agrimonia pilosa Ledebour)




MRC-5




4 4 Mountain leek ( Allium thunbergii G.Don) 4 3 Poria cocas Wolf) 3 4 Acer tegmentosum Maximowocz) 4 4 Paeonia suffruticosa ) 3 3 Golden ( Scutellaria baicalensis Georgi) 3 4 Artemisia annua Linne) 4 4 Mixture (Seokhogcho + Mountain Leek + Baekbongseong + Socheongmyeong + Dogwood Mugwort) 4 4 virus VZV VZV Acyclovir (RFID) 4 4 Echinacea 4 4 Tamiflu 4 4 Saline (Control) 0 0

Example 3. Antiviral assay using MTT method by MTT method

(1) MTT assay

The cells were placed in a 48-well microtiter plate at a cell concentration of 5 × 10 ⁴ cells / ㎖ and placed in 500 μl of each well. The herb extracts (Seonghwak, Sungchul, Baekwolryeong, And then the virus was cultured for 48 hours. The MTT labeling mixture prepared by adding EC (electron coupling reagent) to MTT labeling reagent was treated with 10 μl of each well (final concentration 0.5 mg / ml) for 1 hour, The cytotoxicity was investigated using absorbance at nm wavelength.

(2) Cytoprotective effect of herb extracts on cytotoxicity induced by herpes zoster virus

To investigate the cytoprotective effect of herpesia causing herpes zoster virus (HIV-1, HIV-2 and VZV) causing cell damage in Vero cells, MTT assay, a cytotoxicity indicator, All herbal extracts (ethanol extraction, hot water extraction) extracts were able to confirm the cytoprotective effect. (Figs. 5 to 7, ** P < 0.01, * P < 0.005).

Production Example 1. Anti-herpes virus skin-lotion cosmetic composition containing herbal extracts

An example of the preparation of an anti-virus skin / lotion (100 ml) cosmetic composition containing an herbal extract according to the present invention is as follows.

ingredient Content (% by weight) Purified water 87.188 Modified alcohol 4.000 Seokhakcho + mountain leek + Baekbongseong + sancheongmyeong + white wormwood 4.000 Hyaluronic Acid 2.000 Dipropylene glycol 0.500 glycerin 0.500 Panthenol 0.500 Betaine 0.500 Fiji-60 Hydrogenated Castor Oil 0.300 Methylparaben 0.200 Spices 0.080 Propylparaben 0.050 Octyldodec-25 0.050 Acrylate / C10-30 alkyl acrylate crosspolymer 0.050 Triethanolamine 0.050 Tree sodium 0.020 Xanthan gum 0.012 Sum 100,000

Production Example 2. Serum cosmetic composition for anti-herpes virus containing herbal extract

An example of the preparation of anti-virus serum / 30 ml cosmetic composition containing herbal extracts according to the present invention is as follows.

ingredient Content (% by weight) Purified water 83.230 Modified alcohol 3.000 Methylgluses-20 3.000 Seokhakcho + mountain leek + Baekbongseong + sancheongmyeong + white wormwood 4.000 glycerin 1.500 Dipropylene glycol 1.500 Hyaluronic Acid 1,000 Butylene glycol 0.500 Fiji-60 Hydrogenated Castor Oil 0.800 Panthenol 0.500 Acrylate / C10-30 alkyl acrylate crosspolymer 0.300 Triethanolamine 0.300 Methylparaben 0.200 Hydroxypropylcyclodextrin 0.100 Spices 0.100 Propylparaben 0.050 Tree sodium 0.020 Sum 100,000

Production Example 3. Trojan (candy) composition for an antiseptic virus containing an herb extract

An example of the preparation of a troki (candy) composition for an antiseptic virus containing an herbal extract according to the present invention is as follows.

ingredient Content (% by weight) Seokhakcho + mountain leek + Baekrimyeon + dogwood mugwort + golden 100 honey 1500 Vitamin C 50 Vitamin B6 10 Nicotinic acid amide 10 Royal jelly 80 Preservatives, incense, a very small amount Purified water Balance Sum 2000

Claims (10)

An anti-herpes virus agent comprising as an active ingredient any one or more extracts selected from the group consisting of sunhakcho, wild leek, baekbokyeong, sancheongmok, barkwood, golden or firewood. The method of claim 1,
Wherein the herpes zoster virus is a herpes simplex virus type 1, herpes simplex virus type 2, or herpes zoster virus. A pharmaceutical composition for preventing and treating herpes zoster virus infection comprising the anti-herpesvirus agent according to claim 1 or 2 as an active ingredient. The method of claim 3,
At least one additive selected from the group consisting of carriers, excipients, disintegrants, sweeteners, coating agents, swelling agents, lubricants, lubricants, flavors, antioxidants, buffers, bacteriostats, diluents, dispersants, surfactants, Wherein said medicament is for preventing and treating herpes zoster virus infection. The method of claim 3,
Wherein the pharmaceutical composition is selected from the group consisting of granules, powders, coated tablets, tablets, pills, capsules, suppositories, gels, syrups, juices, suspensions, emulsions, &Lt; / RTI &gt; A cosmetic composition for the prevention and treatment of herpes virus infection diseases comprising the anti-herpes virus agent according to claim 1 or 2 as an active ingredient. The method according to claim 6,
Autoimmune infectious diseases further comprising one or more additives selected from the group consisting of diluents, gels, dispersants, vitamins, surfactants, emulsifiers, flavorings, pigments, stabilizers, preservatives, antioxidants, sunscreens, pH adjusters and chelating agents Cosmetic composition for the prevention and treatment of. Food composition for the prevention and treatment of herpes virus infection disease comprising the anti-herpes virus agent according to claim 1 or 2 as an active ingredient. 9. The method of claim 8,
Food composition for the prevention and treatment of herpes virus infection disease further comprising at least one additive selected from the group consisting of organic acids, phosphates, antioxidants, lactose casein, dextrin, glucose, sugar and sorbitol. 9. The method of claim 8,
The food composition is a powder, granules, tablets, capsules or beverages, characterized in that the food composition for the prevention and treatment of herpes virus infection disease.

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