CN111920912A

CN111920912A - A topical composition for preventing and treating influenza

Info

Publication number: CN111920912A
Application number: CN202010736988.3A
Authority: CN
Inventors: 鲍玉; 王玉玲; 李霞; 郭学平; 郭珈均; 邵萌; 王静
Original assignee: Shandong Huaxi Haiyu Biological Medicine Co Ltd
Current assignee: Shandong Huaxi Haiyu Biological Medicine Co Ltd; Shandong Bloomage Hyinc Biopharm Co Ltd
Priority date: 2020-07-28
Filing date: 2020-07-28
Publication date: 2020-11-13

Abstract

The invention discloses an external composition for preventing and treating influenza, which comprises polylysine, ergothioneine and brown rice fermentation filtrate. The composition can be made into liquid preparation, such as spray, solution, gel, liquid containing agent, etc. The composition disclosed by the invention is simple in components and convenient to use, can have a good effect on prevention and treatment of influenza, is not easy to generate drug resistance, has no toxic or side effect and adverse reaction, and also has the effects of moisturizing and nourishing skin and promoting the synthesis of antibacterial peptide to achieve skin microecological balance.

Description

A topical composition for preventing and treating influenza

Technical Field

The invention relates to an external composition for preventing and treating influenza, in particular to an external composition for preventing and treating influenza, which has good effect, is not easy to generate drug resistance, has no toxic or side effect and adverse reaction.

Background

Influenza (referred to as influenza for short) is a common acute respiratory infectious disease caused by influenza virus. It features short incubation period, fast transmission speed and high disease rate, and the patients show sudden fever, pharyngalgia, dry cough, hypodynamia, redness of bulbar conjunctiva and general muscular soreness. General discomfort lasts for several days, and viral pneumonia or secondary infection of the lung can be caused when the general discomfort lasts for several days.

On one hand, influenza belongs to self-limiting diseases in Western medicine, and no exact anti-pathogen medicine exists at present. The specific anti-influenza virus drugs for controlling and preventing influenza etiology mainly comprise: amantadine, rimantadine, ribavirin, oseltamivir, zanamivir and the like, and the treatment should be used within 48h of the onset of disease. However, all western medicines for treating cold aim at relieving symptoms, shortening the course of disease and preventing and treating complications, and generally have the defects of relatively obvious toxic and side effects and easy generation of drug resistance, and cannot be used for preventing influenza.

From the perspective of traditional Chinese medicine, cold and influenza are caused by external wind-cold on the basis of internal fire in the body. The traditional Chinese medicine for treating cold can clear internal heat, dispel exterior wind-cold evil, treat exterior and interior simultaneously, and can integrally regulate the immune function of human body to achieve the purposes of strengthening body resistance and eliminating evil. However, many of the traditional Chinese medicine compositions have many disadvantages, for example, the patent with the application number of 201110241373.4 discloses a composition for preventing and treating cold and a preparation method thereof, the composition is prepared by ginseng, Chinese date, dried ginger, wild chrysanthemum flower, tea, medlar, longan, astragalus root, liquorice and brown sugar according to a certain proportion, the composition has obvious effect, but has the disadvantages of many components and complicated preparation. For example, the application No. 201510917926.1 discloses a traditional Chinese medicine composition for preventing influenza and an application thereof, wherein the traditional Chinese medicine composition is prepared from the following raw material medicines: radix Angelicae Dahuricae, rhizoma Atractylodis, rhizoma Acori Graminei, herba Agastaches, fructus Arctii, herba Eupatorii, folium Perillae, folium Artemisiae Argyi, Borneolum Syntheticum, and flos Chrysanthemi. The composition is convenient to use after being prepared into a sachet, but has the defects of unobvious effect, multiple components and the like.

Therefore, the existing product has the defects of unobvious effect, more components, toxic and side effects on human bodies and the like when used for preventing influenza. Therefore, it is urgently needed to develop a product with definite curative effect, simple components and no harm.

Polylysine (-PL) is a homo-type monomer polymer containing 25-35L-lysine residues produced by fermentation of Streptococcus albus (Streptomyces albulus), and is formed by connecting an amino group of L-lysine and an alpha-carboxyl group of another L-lysine through an amide bond. The structural formula is as follows:

polylysine has very good safety. Can be decomposed into lysine in human body, and the lysine is one of 8 amino acids necessary for human body, and its acute oral toxicity is 5g/kg, and its research proves thatPL is non-toxic to reproduction, nervous system, immune function, embryonic development and the like.

Because polylysine has a wide antibacterial spectrum, the polylysine has obvious inhibiting and killing effects on candida acutifolia, rhodotorula farinosa, pichia membranaceus, rhodotorula rosea, heat-resistant bacillus steatovora, bacillus coagulans and bacillus subtilis in gram-positive bacteria, arthrobacter aerogenes, escherichia coli and the like in gram-negative bacteria, and therefore, the polylysine is widely used as a cosmetic and food preservative. However, no virus inhibition by polylysine has been reported.

Disclosure of Invention

Aiming at the defects of multiple components, complex preparation, easy generation of drug resistance and the like of the existing drugs for treating or preventing influenza, the invention provides the external composition for preventing and treating influenza, which is an external product, has simple components, no toxic or side effect, difficult generation of drug resistance and good effect on preventing and treating influenza.

The research of the invention finds that-polylysine has better anti-influenza virus effect, especially has better effect after being combined with ergothioneine and brown rice fermentation filtrate, and has better prevention and treatment effect on influenza. Meanwhile, researches also find that the combination of polylysine, ergothioneine and brown rice fermentation filtrate also has the effects of moisturizing and nourishing the skin, can promote the synthesis of antibacterial peptide to achieve the microecological balance of the skin, reduce the invasion of harmful bacteria to organisms and further play a role in preventing cold.

The invention provides an external composition for preventing and treating influenza, which comprises the following effective components in percentage by weight: 0.005-1.0% of polylysine, 0.00002-0.0021% of ergothioneine and 0.1-10% of brown rice fermentation filtrate.

Preferably, the external composition comprises the following effective components in percentage by weight: 0.05 to 0.5 percent of polylysine, 0.0002 to 0.0011 percent of ergothioneine and 1 to 5 percent of brown rice fermentation filtrate.

Furthermore, the polylysine is a homotypic monomer polymer polylysine containing 25-35L-lysine residues.

Furthermore, Ergothioneine (L-Ergothionine, EGT) is the only known natural 2-thioimidazole amino acid with the chemical name of 2-mercaptohistidine trimethyl inner salt, and is mainly used in cosmetics as an antioxidant component and an anti-inflammatory component at present. According to the invention, the experiment shows that the combination of polylysine, ergothioneine and brown rice fermentation filtrate has good prevention and treatment effects on influenza.

At present, many methods for preparing ergothioneine are reported, including a chemical synthesis method, an extraction method and a biological fermentation synthesis method, and the fermentation method is low in cost, convenient for industrial production and the main production method at present. Ergothioneine prepared by various methods can be applied to the invention, and the ergothioneine can exist in a pure form, and also can exist in a fermentation solution or an extraction solution. Preferably, the ergothioneine used in the present invention is ergothioneine obtained by fermentation, for example, ergothioneine matsutake mushroom extract produced by the company itself, which is a fermentation liquid obtained by fermenting matsutake mushroom as a raw material, and contains components such as glucan, polypeptide, amino acid, polysaccharide, etc. in addition to ergothioneine (the content of ergothioneine is 200mg/L or more).

Further, the brown rice fermentation filtrate is obtained by performing impurity removal and sterilization treatment on fermentation liquor obtained by fermenting brown rice with yeast, and the more specific preparation method is as follows: accelerating germination of brown rice, crushing the brown rice into brown rice powder, preparing the brown rice powder into rice pulp, performing enzymolysis on the rice pulp, adding yeast for fermentation, and removing impurities and bacteria from fermentation liquor to obtain brown rice fermentation filtrate. The brown rice is whole grain rice with complete vitality, contains rich dietary fiber, functional active ingredients and mineral substances, and can improve cell activity and human body immunity. Fermenting brown rice with yeast, fully releasing, converting and enriching nutrients in the fermented brown rice, wherein the obtained brown rice fermentation filtrate contains rich active ingredients such as gamma-aminobutyric acid, polypeptide, amino acid, active oxygen resistant phytic acid, ferulic acid, lactic acid, plant amide and sterol, and the amino acid types comprise more than 20 of hydroxyproline, aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, ornithine, lysine, histidine, tryptophan and arginine. The main effects of the brown rice fermentation filtrate reported at present are to improve cell activity, promote cell regeneration and tissue repair, improve skin darkness and make skin moist and beautiful. The brown rice fermentation filtrate is a product produced by the company, and the content of the gamma-aminobutyric acid in the product is as high as 1 g/L.

Further, in a specific embodiment of the present invention, there is provided an external composition for preventing influenza, which comprises the following effective ingredients in percentage by weight: 0.005-1.0% of polylysine, 0.1-10% of ergothioneine matsutake extract and 0.1-10% of brown rice fermentation filtrate. Wherein the content of ergothioneine in the extract of Armillaria mellea is 207 mg/L.

Further, in a specific embodiment of the present invention, there is provided an external composition for preventing influenza, which comprises the following effective ingredients in percentage by weight: 0.05-0.5% of polylysine, 1-5% of ergothioneine matsutake extract and 1-5% of brown rice fermentation filtrate. Wherein the content of ergothioneine in the extract of Armillaria mellea is 207 mg/L.

Furthermore, the external composition may also comprise effective components with bactericidal, antibacterial, and antiviral effects, such as one or more of flos Lonicerae extract, fructus Zanthoxyli extract, benzalkonium chloride, polyhexamethylene biguanide, and ethanol.

Further, the composition for external use may further contain an active ingredient having an action of repairing, moisturizing or the like, for example, ectoin or the like.

Furthermore, the external composition may further include adjuvants required for preparing various feasible preparations, such as one or more of moisturizer, pH regulator, viscosity regulator, perfume, pigment, surfactant and solvent. These adjunct ingredients may be selected from the adjuvants available in the medical field, for example the humectant is selected from one or more of glycerol, sorbitol, betaine, allantoin, acetyl chitosamine, beta-glucan, but not limited thereto. The pH regulator is selected from one or more of citric acid, sodium hydroxide, triethanolamine and aminobenzyl alcohol. The viscosity regulator is selected from one or more of sodium chloride, starch, xanthan gum and cellulose. The surfactant is selected from one or more of polyoxyethylene polyoxypropylene ether, K12 ammonium salt, ammonium laureth sulfate, sodium laureth sulfate and cocamidopropyl betaine.

Further, the above composition for external use may be formulated into various dosage forms, preferably into liquid preparations such as spray, solution, gel, liquid, etc.

Further, the pH of the composition for external use of the present invention is in the range of 5.0 to 7.5.

Furthermore, the preparation method of the external composition is simple, and the external composition is prepared by selecting the required raw materials and preparing the raw materials according to the conventional preparation method of each dosage form disclosed in the prior art.

In one embodiment of the present invention, a specific preparation method of a gel formulation is disclosed, which comprises the following steps: uniformly stirring and dispersing the glycerol, the butanediol and the hydroxyethyl cellulose according to the prescription amount, adding water, heating to 80-95 ℃, and stirring for dissolving. Cooling to 30-40 deg.C after completely dissolving, adding formula amount of polylysine, ergothioneine, and brown rice fermentation filtrate, stirring to dissolve, and adjusting pH to 5.0-7.5 with appropriate amount of sodium hydroxide to obtain gel.

In one embodiment of the present invention, a specific preparation method of a spray formulation is disclosed, which comprises the following steps: weighing the formula amount of glycerol, -polylysine, ergothioneine, brown rice fermentation filtrate, benzalkonium chloride and ectoin, adding into the balance of water, stirring for dissolving, adjusting pH to 5.0-7.5 with appropriate amount of sodium hydroxide after complete dissolution, and filling into a spray bottle to obtain the spray.

In one embodiment of the present invention, a specific preparation method of a mouthwash formulation is disclosed, comprising: dissolving polylysine, ergothioneine and brown rice fermentation filtrate in appropriate amount of water; taking the prescription amount of glycerol and xylitol, and adding the glycerol and the xylitol into the prepared aqueous solution; taking menthol and poloxamer 407 with the prescription amount, adding a proper amount of water, stirring for dissolving, then adding the mixture into the aqueous solution, finally adjusting the pH to 5.0-7.5 by using a pH regulator, adding water for complementing 100%, and stirring and uniformly mixing to obtain the mouthwash.

The external composition is convenient to use, can be dripped or sprayed into a nasal cavity, can be sprayed or contained in an oral cavity, can be sprayed or smeared on hands, and can be sprayed on a mask.

The invention has the following beneficial effects:

1. the polylysine, the ergothioneine and the brown rice fermentation filtrate are compounded to form a layer of biomembrane on the surface of the skin or the respiratory mucosa, so that external bacteria and viruses are isolated, bacterial viruses adhered to the surface of the skin or the respiratory mucosa can be killed, the resistance of the polylysine to the viruses is increased, and influenza is effectively prevented and treated.

2. The synergistic compounding of the polylysine, the ergothioneine and the brown rice fermentation filtrate can promote the synthesis of the antibacterial peptide so as to keep the microecological balance of the skin and reduce the invasion of harmful bacteria to organisms, and meanwhile, the synergistic compounding of the polylysine, the ergothioneine and the brown rice fermentation filtrate can also enhance the wettability of the skin and respiratory mucosa, promote the movement of nasal cilia and the like, and is beneficial to quickly discharging bacterial viruses out of the nasopharynx.

3. Polylysine, ergothioneine and brown rice fermentation filtrate are all biological sources, have safe components and simple components, have good effect on preventing and treating influenza, are not easy to generate drug resistance, and have no toxic or side effect or adverse reaction.

4. The invention has simple preparation and convenient use, can be applied to nasal cavities, oral cavities, hands and other parts in ways of smearing, spraying, dripping, gargling and the like, can also be applied to masks, and can achieve good effects of preventing and treating influenza by external application.

Detailed Description

For better understanding of the present invention, the following examples are given for illustrative purposes and are not intended to limit the scope of the present invention.

The experimental methods used in the following examples are all conventional methods, unless otherwise specified.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

In the examples described below, the ergothioneine used was an ergothioneine Armillaria matsutake extract containing 207mg/L of ergothioneine (batch No. 20040231) from Huaxi Biotech Ltd.

In the following examples, the brown rice fermentation filtrate used was obtained from Huaxi Biotech Co., Ltd, the content of gamma-aminobutyric acid was 1.1g/L, and the product lot was 1812034.

Examples 1 to 5 and comparative examples 1 to 8

Weighing polylysine, ergothioneine Armillaria mellea extract and brown rice fermentation filtrate according to the following table 1, dissolving polylysine in the ergothioneine Armillaria mellea extract and brown rice fermentation filtrate, mixing, stirring, dissolving completely, supplementing 100g with water to obtain composition, and storing in dark place.

Test example 1 test for promoting production of antibacterial peptide in skin

The experiment was conducted using keratinocytes in the logarithmic growth phase, which were cultured in KSFM culture medium containing the compositions of examples 1-5 and comparative examples 1-8 for 24 hours, respectively, as an experimental group, and a control group containing no composition, each at a concentration of 200. mu.g/mL.

Cathelicidin LL37 is the most common AMPs in human skin, and the expression of Cathelicidin LL37mRNA and protein in keratinocytes is detected by expressing the content of Cathelicidin LL37mRNA as the ratio of the IOD value of the Cathelicidin LL37 product to the IOD value of beta-actin, and expressing the content of Cathelicidin LL37 protein as the ratio of the gray scale values of Cathelicidin LL37 to beta-actin.

RT-PCR detection of keratinocyte LL37mRNA expression: the density is about 2 xl 0⁵The cells are inoculated in a 6-well culture plate for 24 hours, so that the cells are attached to the wall. After discarding the culture solution, the composition of the experimental group and the control group was used for intervention, and after washing with PBS, the cells were collected, total RNA was extracted with reference to Trizol instructions, and the a value was determined. First, the cDNA was reverse-transcribed, and then PCR amplification reaction was carried out using the cDNA as a template, the total reaction volume being 20. mu.L. Primer sequences for LL37 were designed: the upstream primer 5'-TCACCAGAGGATTGTGACTTCAA-', the downstream primer 5'-TGAGGGTCACTGTCCCCATAC-'3, the amplification condition is pre-denaturation at 95 ℃ for 2 min, denaturation at 94 ℃ for 20 s, annealing at 57 ℃ for 40 s, extension at 72 ℃ for 40 s, total circulation for 31 times, and extension at 72 ℃ for 8 min. The expected fragment of the amplification product is 547 bp. The upstream primer of the beta-actin internal reference is 5'-ATTGCCGACAGGATGCAGAA-'3, and the downstream primer is 5'-GCTGATCCACATCTGCTGGA-' 3; the amplification conditions were the same as LL37, and the expected fragment of the amplification product was 152 bp. Mixing 5 mul of target gene and internal reference product, performing electrophoresis with 2% agarose gel 100V under constant voltage for 1 h, performing EB staining, and taking a picture by using a gel imaging system. Accumulation of product was determined using Image-Pro Plus 6.0 analytical softwareOptical Density value (IOD), the amount of Cathelidin LL37mRNA expressed as the ratio of the IOD value of Cathelicidin LL37 product to the IOD value of beta-actin.

Western blot to detect the expression of keratinocyte LL37 protein: the density is about l × l0⁶The cells were inoculated in a 10 cm dish for 24h per mL to allow the cells to adhere. After discarding the culture solution, intervening with the composition of the experimental group and the control group, discarding the culture solution after intervening, washing with PBS for 3 times, adding cell lysate containing PMSF, acting on ice for 20min, centrifuging at 12000 r/min for 20min, taking the supernatant, and performing protein quantification by using a BCA method. Loading 30 mu g of total protein into an equal amount of loading buffer solution from each lane, performing SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) separation, and transferring the protein onto a PVDF (polyvinylidene fluoride) membrane; 5% skimmed milk powder is sealed for 1 h at room temperature, primary antibody is added, incubation is carried out overnight at 4 ℃, then horseradish peroxidase-labeled secondary antibody is added, incubation is carried out for 1 h at room temperature, finally ECL color developing agent is added, reaction is carried out for 2 min, color development is carried out, semi-quantitative analysis is carried out by ImageeQuant analysis software, and the content of Catelicidinll 37 protein is expressed by the ratio of Catelicidinll 37 and beta-actin grey values.

The experiment was repeated 3 times and statistical analysis was performed using SPSS 17.0 software.

As can be seen from Table 2, the expression levels of Cathelicidin LL37mRNA and protein were increased when polylysine was used alone, and the expression levels of Cathelicidin LL37mRNA and protein were not increased when the ergothioneine was used alone with the agaricus extract and the brown rice fermented filtrate, indicating that polylysine promotes the production of antibacterial peptides on the skin, but the ergothioneine was not produced with the agaricus extract and the brown rice fermented filtrate. When polylysine and the ergothioneine agaricus extract or the brown rice fermentation filtrate are used in a combined mode, compared with the case that polylysine is used alone, the expression level of Cathelicidin LL37mRNA and protein is improved to some extent, especially when the ergothioneine agaricus extract, the brown rice fermentation filtrate and the polylysine are used in a combined mode, the expression level of Cathelicidin LL37mRNA and protein is improved remarkably, and the fact that the capacity of polylysine for promoting skin to generate the antibacterial peptide can be greatly improved when the ergothioneine agaricus extract and the brown rice fermentation filtrate are used in a combined mode. As is apparent from the comparison between examples 1 to 5 and comparative examples 7 and 8, the expression levels of Cathelidin LL37mRNA and protein were not improved when the content of polylysine in the composition was less than 0.005%, the content of ergothioneine in the Armillaria mellea extract was less than 0.1%, and the expression levels of Cathelidin LL37mRNA and protein were not improved when the content of polylysine in the composition was more than 1.0%, the content of ergothioneine in the Armillaria mellea extract was more than 10%, and the content of brown rice fermentation filtrate was more than 10%, but was lower than that of examples 1 to 5. Therefore, the contents of examples 1 to 5 of the present invention have an obvious synergistic effect based on the combined consideration of cost and effect.

Verification example 2 in vitro antiviral Effect

Influenza A1, influenza A3, adenovirus 3 and adenovirus 7 were diluted 100-fold, mixed with the same amount of the compositions of examples 1-5 and comparative examples 4-6, inoculated into human embryonic kidney cells for adsorption for 40min, and added with a milk protein cell maintenance solution. Simultaneously arranging two tubes of virus and cell reference substances, culturing in an incubator with 37 ℃ and 5% CO2 for 1 week, and observing cytopathic condition, wherein the cytopathic condition is judged according to the following standards:

the test results are shown in Table 3 below.

As is clear from the results shown in Table 3, the ergothioneine Armillaria matsutake extract and the brown rice fermentation filtrate themselves had no action against the above four viruses, and only polylysine had an action against the four viruses, so that the action of polylysine against the four viruses was greatly enhanced by the addition of the ergothioneine Armillaria matsutake extract and the brown rice fermentation filtrate.

Proof example 3 nasal cilia test

1. Instrument and animal

Optical microscopy (Olympus); the weight of the Bufo bufo gargarizans Cantor is 40-50 g, and the Bufo bufo gargarizans Cantor can be used for male and female.

2. Experimental methods

The healthy bufonid 35 were randomly divided into 7 groups, i.e., the composition group of example 5, the composition group of comparative example 1, the composition group of comparative example 2, the composition group of comparative example 3, the physiological saline group, the ephedrine hydrochloride nasal drops group, the sodium deoxycholate solution (10 mg/ml) group, and 5 per group. The Bufo bufo gargarizans Cantor is fixed on the frog board in supine position, and the oral cavity is opened and fixed with hemostatic forceps. Dropping each group of samples on the oral mucosa surface, separating mucosa with the area of about 3mm multiplied by 3mm from the upper jaw mucosa by surgical scissors after 30min, washing blood clots and mucus with normal saline, spreading the mucosa face up on a glass slide, slightly covering the glass slide, and observing the cilia swing condition and the mucosa morphological change under an optical microscope with the power of 40 multiplied by 10 at the ambient temperature of 25 ℃. Observing cilia oscillation condition of mucosa, placing in a chromatographic cylinder with small amount of distilled water to keep mucosa at certain humidity, taking out samples at regular intervals, observing with microscope, and returning to the chromatographic cylinder if cilia movement is continuous. Repeating the observation until the ciliary movement is stopped, recording the time from the start of administration to the stoppage of the ciliary movement, washing the liquid medicine on the mucosa with physiological saline, continuously observing whether the ciliary movement is recovered, and recording the time for recovering the ciliary movement. The relative percentage of ciliary duration was calculated as the ratio of the ciliary duration of the drug-administered fluid group to the ciliary duration of the saline control group. The results of the experiments are shown in Table 4 below.

The mucous membranes of each group are observed under a microscope, the cilia of the normal saline group and the group containing the composition of example 5 are clear and complete, and the cilia move actively; the cilia of the sodium deoxycholate group stopped swinging, and the cilia of the ephedrine hydrochloride nasal drop group and the composition group containing the comparative examples 1-3 were clear and the cilia moved actively. When observation is continued, cilia of the deoxycholate sodium group basically fall off, and only the exposed base part is left to show that the nasal mucosa is seriously damaged; the mucociliary body of the group containing the composition of example 5 was kept moving and stopped for a longer time than the physiological saline group and the ephedrine group, indicating that the composition provided by the present invention can promote the movement of the cilia of the nasal cavity and help to rapidly discharge the bacterial virus out of the nasopharynx.

Verification example 4 evaluation of safety

In vitro cytotoxicity: samples of the compositions of examples 1-5 were each diluted to 1mg/mL in cell culture and tested according to the method of GB/T16886.5-2003, part 5, in vitro cytotoxicity test for medical device biology evaluation. The sample test solutions were contacted with the cultured cells at 37 ℃. + -. 2 ℃ for 48 hours, respectively, and the evaluation criteria are shown in Table 5 below.

Irritation: samples of the compositions of examples 1-5 were each subjected to the following protocol according to GB/T16886.10-2005 Biometrics evaluation part 10: stimulation and delayed hypersensitivity tests.

Sensitization: samples of the compositions of examples 1-5 were each subjected to the following protocol according to GB/T16886.10-2005 Biometrics evaluation part 10: stimulation and delayed type hypersensitivity tests the maximum dose sensitization test method specified in the test.

The results of the safety tests are shown in Table 6 below.

The test results show that the composition of the invention is relatively safe, non-irritant and allergenic.

Example 6

1. Weighing the raw materials according to the formula of the following table 7:

2. preparation of a mouthwash sample: dissolving polylysine, ergothioneine Armillaria mellea extract and brown rice fermentation filtrate in appropriate amount of water; taking the prescription amount of glycerol and xylitol, and adding the glycerol and the xylitol into the prepared aqueous solution; taking menthol and poloxamer 407 with the prescription amount, adding a proper amount of water, stirring for dissolving, then adding the mixture into the aqueous solution, finally adjusting the pH to 5.0-7.5 by using a pH regulator, adding water for complementing 100%, and stirring and uniformly mixing to obtain the mouthwash.

3. Preparing a mouthwash reference: the preparation method of the mouth wash sample was the same except that polylysine, ergothioneine-extracted matsutake extract, and brown rice fermentation filtrate were not added.

The mouthwash sample and the reference substance prepared above are used for influenza prevention tests, and the method is as follows:

selection criteria: susceptible people; ② the elderly;

inclusion criteria were: meeting any one of selection standards; secondly, people who have suffered from common cold are eliminated; ③ no preventive drug or measure is accepted or used; and fourthly, the person who can understand and sign the informed consent.

Exclusion criteria: the medicine has serious cardiovascular and cerebrovascular diseases, liver and kidney diseases, blood diseases, endocrine diseases, lung diseases, neuropsychiatric diseases, diabetes, immunological diseases, chronic hepatitis, AIDS and the like; ② allergic diseases and allergic constitution such as allergic rhinitis and allergic asthma, drug allergy and other acute infectious diseases; ③ alcoholism or other drug abusers; women in pregnancy or lactation period, and chemotherapy period for tumor and other diseases; hyperlipidemia and overeating high fat diet.

100 volunteers meeting the above conditions are collected in the city, wherein 80 volunteers are susceptible and 20 volunteers are old, and the volunteers are randomly divided into two groups, and the two groups have no statistical difference in gender and age and are comparable. The experimental group used mouthwash samples and the control group used mouthwash controls. The test time is 11-12 months, each volunteer is used 3 times per day, once in the morning, in the middle and at night, and the appropriate amount of the medicine is gargled in the mouth for 30 seconds each time, and then the medicine is spitted out. The number of cold people was counted after two consecutive months of use, and the results are shown in table 8 below.

Therefore, the composition has better cold prevention effect.

Example 7

1. Weighing the raw materials according to the formula of the following table 9:

2. preparation of a gel sample: uniformly stirring and dispersing the glycerol, the butanediol and the hydroxyethyl cellulose according to the prescription amount, adding water, heating to 80-95 ℃, and stirring for dissolving. Cooling to 30-40 deg.C after completely dissolving, adding formula amount of polylysine, ergothioneine Armillaria matsutake extract, and brown rice fermentation filtrate, stirring to dissolve, and adjusting pH to 5.0-7.5 with appropriate amount of sodium hydroxide to obtain gel.

3. Preparation of gel reference: the gel sample preparation method was identical except that no polylysine, ergothioneine matsutake extract, and brown rice fermentation filtrate were added.

The gel is used for influenza treatment tests, and the method comprises the following steps:

150 influenza patients were selected as subjects for analysis in this study, and the symptoms of influenza were: sore throat, nasal obstruction, watery nasal discharge, cough, sneeze, headache, muscle pain, aversion to cold, etc. The average age of the patients is 30.8 +/-4.1 years, the course of disease is 1-2 weeks, 75 cases are male and 75 cases are female.

Patients were divided on average into 5 groups with no statistical difference in gender, age and course, four groups being test groups and one group being control group. Gel samples 1-4 were used in the test group and gel control was used in the control group. Each sample and control were applied by squeezing into the nasal cavity, 4-5 drops per nostril at a time, 1-2 hours 1 time, at least 6 times per day, and 7 days. After 7 days, evaluation was performed using the following evaluation criteria.

The efficacy evaluation criteria were as follows:

the effect is shown: the adverse symptoms of influenza patients are obviously improved;

the method is effective; the adverse symptoms of influenza patients are improved;

invalid; the adverse symptoms of influenza patients do not improve or worsen.

Total effective rate = (number of effective cases + number of effective cases)/total number of cases × 100%.

The results of the experiment are shown in table 10 below.

From the above results, it is understood that the polylysine, the ergothioneine matsutake mushroom extract, and the brown rice fermentation filtrate composition have a good effect of improving influenza symptoms, and are effective in treating influenza.

Although the invention has been described in detail above with reference to a general description and examples, it will be apparent to one skilled in the art that modifications and improvements can be made based on the invention. Accordingly, it is intended that all such modifications and variations be included within the scope of the invention as claimed and not departing from the spirit thereof.

Claims

1. A composition for external use for preventing and treating influenza, which is characterized in that: comprises the following effective components in percentage by weight: 0.005-1.0% of polylysine, 0.00002-0.0021% of ergothioneine and 0.1-10% of brown rice fermentation filtrate.

2. The composition for external use according to claim 1, wherein: comprises the following effective components in percentage by weight: 0.05 to 0.5 percent of polylysine, 0.0002 to 0.0011 percent of ergothioneine and 1 to 5 percent of brown rice fermentation filtrate.

3. The composition for external use according to claim 1 or 2, wherein: the ergothioneine is derived from ergothioneine matsutake mushroom extract, and the ergothioneine matsutake mushroom extract is fermentation liquid obtained by fermenting matsutake mushroom as raw material.

4. The composition for external use according to claim 1 or 2, wherein: the brown rice fermentation filtrate is obtained by performing impurity removal and sterilization treatment on fermentation liquor obtained by fermenting brown rice with yeast.

5. The composition for external use according to claim 1 or 2, wherein: also comprises effective components with bactericidal, bacteriostatic and antiviral effects.

6. The composition for external use according to claim 5, wherein: the effective components with antibacterial, bacteriostatic and antiviral effects are selected from one or more of flos Lonicerae extract, fructus Zanthoxyli extract, benzalkonium chloride, polyhexamethylene biguanide and ethanol.

7. The composition for external use according to claim 1 or 2, wherein: also comprises auxiliary materials required by the preparation, which comprise one or more of a humectant, an acid-base regulator, a viscosity regulator, a spice, a pigment, a surfactant and a solvent.

8. The composition for external use according to claim 1 or 2, wherein: the dosage form is a liquid preparation.

9. The composition for external use according to claim 1 or 2, wherein: the formulation of the liquid soap is spray, solution, gel or liquid soap.

10. The composition for external use according to claim 1 or 2, wherein: the pH value is 5.0-7.5.