A plant extract and fibronectin composition, preparation method thereof and application thereof in HPV infection

Abstract

The invention discloses a plant extract and fibronectin composition, a preparation method thereof and an application thereof in HPV infection, and relates to the field of biotechnology. The invention provides a composition, wherein the composition is made of a Chinese medicine composition and fibronectin, and the Chinese medicine composition is made of the following components: Curcuma, Artemisia capillaris, Astragalus, Kochia scoparia, Patrinia scabra, Sophora flavescens, Mentha, Lithospermum officinale, Viola yedoensis, Avena sativa, Cnidium monnieri, Houttuynia cordata, Paeonia lactiflora, Ginseng, Licorice, Portulaca oleracea, Pelargonium, Honeysuckle, Gentiana scabra, Dictamni, Coptis chinensis, Scutellaria baicalensis, Aquilaria sinensis. The composition of the invention has a good antibacterial effect, can effectively inhibit the proliferation of Hela cells and H8 cells, inhibit the expression of HPV oncogenes, can inhibit HPV viruses, prevent and treat HPV infections, and the composition of the invention has high safety, low toxicity and side effects, and has broad application prospects.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a plant extract and fibronectin composition, a preparation method thereof, and an application thereof in HPV infection. Background Art

[0002] Human papillomavirus (HPV) is a small double-stranded circular DNA virus belonging to the family Papillomaviridae, with a strict genotype-specific host distribution. According to its pathogenicity, it is divided into high-risk types (HR-HPVs) and low-risk types (LR-HPVs). Common high-risk HPVs include: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68. Low-risk HPVs are mainly associated with genital warts and benign cervical lesions. The two most common HR-HPVs are HPV16 and HPV18. Most LR-HPVs infections disappear over time, while persistent HR-HPVs play a vital role in the development of cervical cancer. HPV is a non-enveloped DNA virus with an icosahedral capsid that enters the basal cells of the stratified epithelium during the proliferation phase through tiny wounds to cause infection. The integration of HPV DNA into the host cell genome is a key factor in the HPV-mediated carcinogenic process, affecting the host genome through the amplification of oncogenes and the destruction of tumor suppressor genes. Among them, E6 and E7 oncogenes play an important role in inducing cervical cell carcinogenesis.

[0003] Chinese patent CN113244357A discloses a microemulsion gel of anti-HPV plant extracts and its preparation method and application. The microemulsion gel includes plant extracts and excipients. The plant extracts include one or more of the following: Curcuma zedoaria extract, eucalyptus extract, Sophora flavescens extract, Cnidium monnieri extract, Stemona extract, and Phellodendron amurense extract. The invention uses natural plant extracts and carbomer as a matrix to prepare a local external preparation with antibacterial, anti-inflammatory, and antiviral effects. The microemulsion gel is used for HPV infection and prevention, bacterial and fungal vaginitis, cervical erosion, and daily care of female private parts. The microemulsion gel prepared by the invention has high safety, good stability, convenient administration, and strong patient compliance; the processing technology is simple, the cost is low, and it is easy to mass produce.

[0004] Chinese patent CN108721449A discloses a gel for inhibiting HPV infection and effectively enhancing resistance to HPV infection. The gel is made of the following raw materials in the following weight ratio: propolis, cnidium monnieri, sophora flavescens, frankincense, saffron, angelica sinensis, chitosan, hyaluronic acid, biological peptides, and biological proteins. The drug combination promotes cell metabolism, thereby enhancing the cell's ability to absorb drugs. The combination of hyaluronic acid, chitosan, biological peptides, and biological proteins can effectively moisturize the skin and form a thin film on the skin surface, effectively blocking the invasion of foreign bacteria, and quickly repairing skin tissue, improving wound healing and regeneration ability. The various plant extracts in the gel can also improve the body's immunity and have a good antiviral effect.

[0005] Currently, there is little research on therapeutic drugs for HPV infection. Developing more ideal drugs for treating HPV infection, especially safe and effective combinations, is an urgent problem that needs to be solved. Summary of the invention

[0006] The purpose of the present invention is to provide a plant extract and fibronectin composition, a preparation method thereof and an application thereof in HPV infection.

[0007] To achieve the above-mentioned object of the invention, the technical solution of the present invention is as follows:

[0008] On the one hand, the present invention provides a composition, which is made of a traditional Chinese medicine composition and fibronectin, and the traditional Chinese medicine composition is made of the following components: Curcuma, Artemisia capillaris, Astragalus, Kochia scoparia, Patrinia scabra, Sophora flavescens, Mentha, Lithospermum officinale, Viola yedoensis, Avena sativa, Cnidium monnieri, Houttuynia cordata, Paeonia lactiflora, Ginseng, Licorice, Portulaca oleracea, Pelargonium, Honeysuckle, Gentiana scabra, Dictamni, Coptis chinensis, Scutellaria baicalensis, and Aquilaria sinensis.

[0009] Specifically, the Chinese medicine composition is made of the following components by weight: 5-10 parts of Curcuma, 1-4 parts of Artemisia capillaris, 10-15 parts of Astragalus, 6-10 parts of Kochia scoparia, 8-12 parts of Patrinia, 1-4 parts of Sophora flavescens, 3-6 parts of Mentha, 1-3 parts of Lithospermum officinale, 8-12 parts of Viola yedoensis, 1-8 parts of Avena sativa, 10-18 parts of Cnidium monnieri, 1-8 parts of Houttuynia cordata, 10-15 parts of Red Peony Root, 4-8 parts of Ginseng, 5-8 parts of Licorice, 8-12 parts of Portulaca oleracea, 0.1-1.5 parts of Pelargonium, 1-3 parts of Honeysuckle, 5-10 parts of Gentiana, 1-4 parts of Dictamni, 5-15 parts of Coptis chinensis, 1-4 parts of Scutellaria baicalensis, and 5-8 parts of Aquilaria sinensis.

[0010] Furthermore, the Chinese medicine composition is made of the following components by weight: 6-9 parts of Curcuma, 1-3 parts of Artemisia capillaris, 12-14 parts of Astragalus, 7-9 parts of Kochia scoparia, 9-11 parts of Patrinia, 2-4 parts of Sophora flavescens, 4-6 parts of Mentha, 2-3 parts of Lithospermum officinale, 10-12 parts of Viola yedoensis, 4-6 parts of Avena sativa, 12-16 parts of Cnidium monnieri, 5-8 parts of Houttuynia cordata, 10-14 parts of Red Peony Root, 5-8 parts of Ginseng, 6-8 parts of Licorice, 8-10 parts of Portulaca oleracea, 0.5-1.5 parts of Pelargonium, 2-3 parts of Honeysuckle, 5-8 parts of Gentiana scabra, 1-3 parts of Dictamni Cortex, 8-12 parts of Coptis chinensis, 1-3 parts of Scutellaria baicalensis, and 5-7 parts of Aquilaria sinensis.

[0011] Furthermore, the Chinese medicine composition is made of the following components by weight: 8 parts of Curcuma, 2 parts of Artemisia capillaris, 13 parts of Astragalus, 8 parts of Kochia scoparia, 10 parts of Patrinia, 3 parts of Sophora flavescens, 5 parts of Mentha, 2.5 parts of Lithospermum officinale, 11 parts of Viola yedoensis, 5 parts of Avena sativa, 14 parts of Cnidium monnieri, 7 parts of Houttuynia cordata, 12 parts of Red Peony Root, 6 parts of Ginseng, 7 parts of Licorice, 9 parts of Portulaca oleracea, 1 part of Pelargonium, 2 parts of Honeysuckle, 6 parts of Gentiana scabra, 2 parts of Dictamni, 10 parts of Coptis chinensis, 2 parts of Scutellaria baicalensis, and 6 parts of Aquilaria sinensis.

[0012] Specifically, the dosage form of the composition is drops, mixtures, tinctures, injections, tablets, powders, oral liquids, capsules, granules, ointments, suspensions, powders, emulsions, solutions, pellets, pills, buccal tablets, lyophilized powder injections, gels, suppositories or aerosols.

[0013] Furthermore, the composition is in the form of a gel.

[0014] Specifically, the composition contains 1-5 parts of fibronectin.

[0015] Furthermore, the composition contains 2-4 parts of fibronectin.

[0016] Furthermore, the composition comprises 2 portions of fibronectin.

[0017] In another aspect, the present invention provides a method for preparing the above-mentioned composition, comprising the following steps:

[0018] (1) Take parts by weight of Curcuma zedoaria, Astragalus, Sophora flavescens, Lithospermum officinale, Viola yedoensis, Cnidium monnieri, Houttuynia cordata, Coptis chinensis, and Aquilaria sinensis, grind them, add ethanol to perform reflux extraction, filter and concentrate, recover ethanol, and obtain a concentrated solution 1;

[0019] (2) taking parts by weight of Artemisia capillaris, Kochia scoparia, Patrinia scabra, Mentha, Oat kernel, Red Peony Root, Ginseng, Licorice, Portulaca oleracea, Geranium, Honeysuckle, Gentiana scabra, Dictamni Cortex, and Scutellaria baicalensis, adding water for decoction to obtain a decoction, and concentrating to obtain a concentrate 2;

[0020] (3) The concentrated solution 1, the concentrated solution 2 and the fibronectin are mixed to obtain a composition.

[0021] Specifically, the amount of ethanol added in step (1) is 8-12 times the mass of the traditional Chinese medicine.

[0022] Specifically, the volume fraction of ethanol in step (1) is 70-90%.

[0023] Specifically, the number of reflux extractions in step (1) is 1-3 times, and the time for each reflux extraction is 1-3 hours.

[0024] Specifically, the temperature of the reflux extraction in step (1) is 45-60°C.

[0025] Specifically, the relative density is 1.25-1.30 when concentrated to 65°C in step (1).

[0026] Specifically, the amount of water added in step (2) is 8-14 times the mass of the traditional Chinese medicine.

[0027] Specifically, the number of decoctions in step (2) is 1-3 times, and the decoction time for each time is 2-4 hours.

[0028] Specifically, in step (2), the concentration is performed to a relative density of 1.2-1.3.

[0029] In another aspect, the present invention provides use of the above-mentioned composition in the preparation of a medicament for preventing or treating HPV infection.

[0030] Specifically, the drug can be used to prevent or treat vaginitis, cervical erosion, and cervical cancer caused by HPV infection.

[0031] Furthermore, the vaginitis includes bacterial vaginitis and fungal vaginitis.

[0032] Specifically, the medicine may also include pharmaceutically acceptable excipients.

[0033] Furthermore, the pharmaceutically acceptable excipient is selected from one or more of excipients, buffers, emulsifiers, stabilizers, diluents, adhesives, preservatives, lubricants, pH regulators, cryoprotectants, and flavoring agents.

[0034] Specifically, the excipient is selected from at least one of microcrystalline cellulose, lactose, pregelatinized starch, cyclodextrin, carboxymethyl cellulose, mannitol, magnesium stearate, starch, calcium phosphate, ethyl cellulose, methyl cellulose, alginic acid, gelatin, gum arabic, glyceryl monostearate, sodium starch glycolate, guar gum, glycerol, and propylene glycol.

[0035] Specifically, the buffer is selected from at least one of sodium dihydrogen phosphate, sodium bicarbonate, ammonium bicarbonate, sodium acetate, citrate, histidine, and succinate.

[0036] Specifically, the emulsifier is selected from at least one of magnesium stearate, zinc stearate, calcium stearate, glyceryl stearate, sorbitan isostearate, sorbitan oleate, glyceryl oleate, and polyglyceryl-3 polyricinoleate.

[0037] Specifically, the stabilizer is selected from at least one of acacia gum, agar, alginic acid, cellulose ether, and carboxymethyl chitosan.

[0038] Specifically, the diluent is selected from at least one of erythritol, mannitol, sorbitol, xylitol, lactose, sucrose, corn starch, potato starch, calcium phosphate, calcium citrate, and crystalline cellulose.

[0039] Specifically, the binder is selected from at least one of ethanol, starch slurry, syrup, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, sodium alginate, and polyvinyl pyrrolidone.

[0040] Specifically, the preservative is selected from at least one of methylparaben, propylparaben, methylparaben, ethylparaben, propylparaben, chlorobutanol, thimerosal, mercuric cyanide, phenoxyethanol, chlorhexidine, benzoic acid, sodium benzoate, chlorocresol, benzalkonium bromide, benzalkonium chloride, and ethylparaben.

[0041] Specifically, the lubricant is selected from at least one of magnesium stearate, stearic acid, sodium chloride, sodium oleate, sodium lauryl sulfate, and poloxamer.

[0042] Specifically, the pH regulator is selected from at least one of citric acid, fumaric acid, succinic acid, tartaric acid, malic acid and ascorbic acid.

[0043] Specifically, the cryoprotectant is selected from at least one of sucrose, glucose, mannitol, fructose, trehalose, dextrose, lactose, glycerol, methanol, ethanol, ethylene glycol, propylene glycol, dimethyl sulfoxide, acetamide or formamide.

[0044] Specifically, the flavoring agent is selected from at least one of sweet orange flavor, vanilla flavor, strawberry flavor, milk flavor, banana flavor, and cherry flavor.

[0045] In another aspect, the present invention provides use of the above composition in preparing an antibacterial product.

[0046] Furthermore, the products include medicines and sanitary disinfection products.

[0047] Specifically, the antibacterial activity is the inhibition of Escherichia coli, Staphylococcus aureus and Candida albicans.

[0048] The beneficial effects of the present invention are:

[0049] The present invention provides a composition comprising a plant extract and fibronectin for treating HPV virus infection. Studies have found that the composition of the present invention has good antibacterial effect, can effectively inhibit the proliferation of Hela cells and H8 cells, inhibit the expression of HPV oncogenes E6 and E7, and has a significant effect on inhibiting HPV virus, preventing or treating HPV infection. The composition of the present invention has high safety, low toxic and side effects, and has broad application prospects. DETAILED DESCRIPTION

[0050] In order to make the technical means, creative features, purpose and efficacy of the present invention easy to understand, the present invention is further explained in conjunction with specific embodiments below, but the following embodiments are only preferred embodiments of the present invention, not all. Based on the embodiments in the implementation mode, other embodiments obtained by those skilled in the art without creative work all belong to the protection scope of the present invention. In the following embodiments, unless otherwise specified, the operating methods used are conventional operating methods, the equipment used are conventional equipment, and the equipment materials used in each embodiment are the same.

[0051] The fibronectin in the following examples and comparative examples was provided by Guangzhou Fiberlink Biotechnology Co., Ltd.

[0052] Example 1 Preparation of a composition for resisting HPV infection

[0053] The components and contents in the composition in Example 1 are:

[0054] 8 parts of Curcuma, 2 parts of Artemisia capillaris, 13 parts of Astragalus, 8 parts of Kochia scoparia, 10 parts of Patrinia, 3 parts of Sophora flavescens, 5 parts of Mentha, 2.5 parts of Lithospermum officinale, 11 parts of Viola yedoensis, 5 parts of Avena sativa seeds, 14 parts of Cnidium monnieri, 7 parts of Houttuynia cordata, 12 parts of Red Peony Root, 6 parts of Ginseng, 7 parts of Licorice, 9 parts of Portulaca oleracea, 1 part of Pelargonium, 2 parts of Honeysuckle, 6 parts of Gentiana, 2 parts of Dictamni, 10 parts of Coptidis, 2 parts of Scutellaria baicalensis, 6 parts of Aquilaria and 2 parts of fibronectin.

[0055] The preparation method of the composition comprises the following steps:

[0056] (1) Take parts by weight of Curcuma zedoaria, Astragalus, Sophora flavescens, Lithospermum officinale, Viola yedoensis, Cnidium monnieri, Houttuynia cordata, Coptis chinensis, and Aquilaria sinensis, grind them, add 8 times the volume fraction of 75% ethanol, perform reflux extraction at 45° C., perform reflux extraction twice, and each extraction time is 2 h. Combine the extracts, filter and concentrate to a relative density of 1.25-1.30 at 65° C., recover the ethanol, and obtain a concentrated solution 1;

[0057] (2) Take parts by weight of Artemisia capillaris, Kochia scoparia, Patrinia scabra, Mentha, Oat kernel, Red Peony Root, Ginseng, Licorice, Portulaca oleracea, Geranium, Honeysuckle, Gentiana scabra, Dictamni Cortex, and Scutellaria baicalensis, add 10 times the amount of water to decoct, decoct for 3 times, each decoction time is 3 hours, combine the decoctions, and concentrate to a relative density of 1.2-1.3 to obtain concentrated 2;

[0058] (3) The concentrated solution 1, the concentrated solution 2 and the fibronectin are mixed to obtain a composition.

[0059] The preparation of the gel comprises the following steps:

[0060] Take 2 parts of carbomer and put it in 85 parts of deionized water to make it swell. After it is completely swollen, add 8 parts of glycerol and stir to form a transparent gel matrix. Add 10 parts of the above-prepared composition and adjust the pH to 4.5-5.5 to obtain the product.

[0061] Example 2 Preparation of a composition for resisting HPV infection

[0062] The components and contents in the composition in Example 2 are:

[0063] 6 parts of Curcuma, 1 part of Artemisia Capillaris, 12 parts of Astragalus, 7 parts of Kochia scoparia, 9 parts of Patrinia, 2 parts of Sophora flavescens, 4 parts of Mentha, 2 parts of Lithospermum officinale, 10 parts of Viola yedoensis, 4 parts of Avena sativa seeds, 12 parts of Cnidium monnieri, 5 parts of Houttuynia cordata, 10 parts of Red Peony Root, 5 parts of Ginseng, 6 parts of Licorice, 8 parts of Portulaca oleracea, 0.5 parts of Pelargonium, 2 parts of Honeysuckle, 5 parts of Gentiana scabra, 1 part of Dictamni Cortex, 8 parts of Coptidis Rhizoma, 1 part of Scutellaria baicalensis, 5 parts of Aquilariae and 2 parts of fibronectin.

[0064] The preparation method of the composition and the preparation method of the gel are the same as those in Example 1.

[0065] Example 3 Preparation of a composition for resisting HPV infection

[0066] The components and contents in the composition in Example 3 are:

[0067] 9 parts of Curcuma, 3 parts of Artemisia Capillaris, 14 parts of Astragalus, 9 parts of Kochia scoparia, 11 parts of Patrinia, 4 parts of Sophora flavescens, 6 parts of Mentha, 3 parts of Lithospermum officinale, 12 parts of Viola yedoensis, 6 parts of Avena sativa seeds, 16 parts of Cnidium monnieri, 8 parts of Houttuynia cordata, 14 parts of Red Peony Root, 8 parts of Ginseng, 8 parts of Licorice, 10 parts of Portulaca oleracea, 1.5 parts of Pelargonium, 3 parts of Honeysuckle, 8 parts of Gentiana scabra, 3 parts of Dictamni, 12 parts of Coptidis, 3 parts of Scutellaria baicalensis, 7 parts of Aquilaria and 4 parts of fibronectin.

[0068] The preparation method of the composition and the preparation method of the gel are the same as those in Example 1.

[0069] Example 4 Preparation of a Combination for Anti-HPV Infection

[0070] The components and contents of the composition in Example 4 are the same as those in Example 1.

[0071] The preparation method of the composition comprises the following steps:

[0072] (1) Take parts by weight of Curcuma zedoaria, Astragalus, Sophora flavescens, Lithospermum officinale, Viola yedoensis, Cnidium monnieri, Houttuynia cordata, Coptis chinensis, and Aquilaria sinensis, grind them, add 10 times the volume fraction of 70% ethanol, perform reflux extraction at 45° C., perform reflux extraction 3 times, each extraction time is 2 h, combine the extracts, filter and concentrate to a relative density of 1.25-1.30 at 65° C., recover the ethanol, and obtain a concentrated solution 1;

[0073] (2) Take parts by weight of Artemisia capillaris, Kochia scoparia, Patrinia scabra, Mentha, Oat kernel, Red Peony Root, Ginseng, Licorice, Portulaca oleracea, Geranium, Honeysuckle, Gentiana scabra, Dictamni Cortex, and Scutellaria baicalensis, add 12 times the amount of water to decoct, the decoction is performed twice, each decoction time is 4 hours, the decoction is combined, and concentrated to a relative density of 1.2-1.3 to obtain concentrated 2;

[0074] (3) The concentrated solution 1, the concentrated solution 2 and the fibronectin are mixed to obtain a composition.

[0075] The preparation of the gel comprises the following steps:

[0076] Take 2 parts of carbomer and put it in 85 parts of deionized water to make it swell. After it is completely swollen, add 8 parts of glycerol and stir to form a transparent gel matrix. Add 12 parts of the above-prepared composition and adjust the pH to 4.5-5.5 to obtain the product.

[0077] Comparative Example 1

[0078] The difference from Example 1 is that the composition of the composition is different. The components and contents in the composition of Comparative Example 1 are as follows:

[0079] 8 parts of Curcuma, 13 parts of Astragalus, 3 parts of Sophora flavescens, 2.5 parts of Lithospermum officinale, 5 parts of Avena sativa, 14 parts of Cnidium monnieri, 7 parts of Houttuynia cordata, 1 part of Pelargonium, 6 parts of Gentiana, 10 parts of Coptidis, 6 parts of Aquilariae and 2 parts of fibronectin.

[0080] The preparation method of the composition comprises the following steps:

[0081] (1) Take parts by weight of Curcuma zedoaria, Astragalus, Sophora flavescens, Lithospermum officinale, Cnidium monnieri, Houttuynia cordata, Coptis chinensis, and Aquilaria sinensis, grind them, add 8 times the volume fraction of 75% ethanol, perform reflux extraction at 45° C., perform reflux extraction twice, and each extraction time is 2 hours. Combine the extracts, filter and concentrate to a relative density of 1.25-1.30 at 65° C., recover the ethanol, and obtain a concentrated solution 1;

[0082] (2) Take oat kernels, geranium and gentian in different weight proportions, add 10 times the amount of water and boil for 3 times, each boiling time is 3 hours, combine the decoctions, and concentrate to a relative density of 1.2-1.3 to obtain concentrated 2;

[0083] (3) The concentrated solution 1, the concentrated solution 2 and the fibronectin are mixed to obtain a composition.

[0084] The preparation of the gel is the same as in Example 1.

[0085] Comparative Example 2

[0086] The difference from Example 1 is that the composition of the composition is different. The components and contents in the composition of Comparative Example 2 are as follows:

[0087] 2 parts of Artemisia Capillaris, 8 parts of Kochia scoparia, 10 parts of Patrinia scabra, 5 parts of Mentha, 11 parts of Viola yedoensis, 6 parts of Ginseng, 9 parts of Portulaca oleracea, 12 parts of Red Peony Root, 2 parts of Scutellaria baicalensis, 2 parts of Honeysuckle, 7 parts of Licorice, and 2 parts of Dictamni.

[0088] The preparation method of the composition comprises the following steps:

[0089] (1) Take a certain amount of Viola yedoensis by weight, crush it, add 8 times the amount of 75% ethanol by volume, perform reflux extraction at 45°C, perform reflux extraction twice, and each extraction time is 2 hours. Combine the extracts, filter and concentrate to a relative density of 1.25-1.30 at 65°C, recover the ethanol, and obtain a concentrated solution 1;

[0090] (2) Take parts by weight of Artemisia capillaris, Kochia scoparia, Patrinia scabra, Mentha, Paeonia lactiflora, Licorice, Portulaca oleracea, Honeysuckle, Dictamni cortex, Scutellaria baicalensis, and Ginseng, add 10 times the amount of water to decoct, the number of decoctions is 3 times, each decoction time is 3 hours, the decoctions are combined, and concentrated to a relative density of 1.2-1.3 to obtain concentrated 2;

[0091] (3) The concentrated solution 1 and the concentrated solution 2 are mixed to obtain a composite material.

[0092] The preparation of the gel is the same as in Example 1.

[0093] Comparative Example 3

[0094] The only difference from Example 1 is the content of the components of the composition. The composition in Comparative Example 3 is:

[0095] 12 parts of Curcuma, 5 parts of Artemisia Capillaris, 16 parts of Astragalus, 12 parts of Kochia scoparia, 14 parts of Patrinia, 5 parts of Sophora flavescens, 8 parts of Mentha, 5 parts of Lithospermum officinale, 14 parts of Viola yedoensis, 10 parts of Avena sativa seeds, 20 parts of Cnidium monnieri, 10 parts of Houttuynia cordata, 16 parts of Red Peony Root, 9 parts of Ginseng, 10 parts of Licorice, 14 parts of Portulaca oleracea, 2 parts of Pelargonium, 4 parts of Honeysuckle, 12 parts of Gentiana scabra, 5 parts of Dictamni, 16 parts of Coptidis Rhizoma, 5 parts of Scutellariae, 10 parts of Aquilariae and 6 parts of fibronectin.

[0096] The preparation method of the composition and the preparation method of the gel are the same as those in Example 1.

[0097] Comparative Example 4

[0098] The only difference from Example 1 is the content of the components of the composition. The composition in Comparative Example 4 is:

[0099] 4 parts of Curcuma, 0.5 parts of Artemisia capillaris, 8 parts of Astragalus, 5 parts of Kochia scoparia, 6 parts of Patrinia, 0.5 parts of Sophora flavescens, 2 parts of Mentha, 0.5 parts of Lithospermum officinale, 6 parts of Viola yedoensis, 0.8 parts of Avena sativa seeds, 8 parts of Cnidium monnieri, 0.8 parts of Houttuynia cordata, 9 parts of Red Peony Root, 3 parts of Ginseng, 4 parts of Licorice, 6 parts of Portulaca oleracea, 0.08 parts of Pelargonium, 0.8 parts of Honeysuckle, 4 parts of Gentiana, 0.8 parts of Dictamni, 4 parts of Coptidis, 0.5 parts of Scutellaria baicalensis, 4 parts of Aquilaria and 0.8 parts of fibronectin.

[0100] The preparation method of the composition and the preparation method of the gel are the same as those in Example 1.

[0101] Comparative Example 5

[0102] The difference between Comparative Example 5 and Example 1 is that Poria is used to replace Artemisia capillaris, Folium Isatidis is used to replace Houttuynia cordata, Fructus Cnidium is used to replace Fructus Forsythiae, Peppermint is replaced by Hedyotis diffusa, and fibronectin is replaced by whey protein.

[0103] The components and contents in the composition of Comparative Example 5 are:

[0104] 8 parts of Curcuma, 2 parts of Poria, 13 parts of Astragalus, 8 parts of Kochia scoparia, 10 parts of Patrinia, 3 parts of Sophora flavescens, 5 parts of Oldenlandia diffusa, 2.5 parts of Lithospermum officinale, 11 parts of Viola yedoensis, 5 parts of Avena sativa, 14 parts of Fructus Forsythiae, 7 parts of Folium Isatidis, 12 parts of Radix Paeoniae Rubra, 6 parts of Ginseng, 7 parts of Liquorice, 9 parts of Portulaca Oleracea, 1 part of Pelargonium, 2 parts of Flos Lonicerae, 6 parts of Gentiana, 2 parts of Cortex Dictamni, 10 parts of Rhizoma Coptidis, 2 parts of Radix Scutellariae, 6 parts of Aquilariae Reticulatae and 2 parts of whey protein.

[0105] The preparation method of the composition comprises the following steps:

[0106] (1) Take parts by weight of Curcuma zedoaria, Astragalus, Sophora flavescens, Lithospermum officinale, Viola yedoensis, Fructus Forsythiae, Folium Isatidis, Rhizoma Coptidis, and Aquilaria sinensis, grind them, add 8 times the volume fraction of 75% ethanol, perform reflux extraction at 45°C, perform reflux extraction twice, and each extraction time is 2 hours. Combine the extracts, filter and concentrate to a relative density of 1.25-1.30 at 65°C, recover the ethanol, and obtain a concentrated solution 1;

[0107] (2) Take parts by weight of Poria cocos, Kochia scoparia, Patrinia scabra, Hedyotis diffusa, Avena sativa, Paeonia lactiflora, Ginseng, Licorice, Portulaca oleracea, Geranium, Honeysuckle, Gentiana scabra, Dictamni, and Scutellaria baicalensis, add 10 times the amount of water to decoct, decoct 3 times, each decoction time is 3 hours, combine the decoctions, and concentrate to a relative density of 1.2-1.3 to obtain concentrated 2;

[0108] (3) The concentrated solution 1, the concentrated solution 2 and the whey protein are mixed to obtain a composition.

[0109] The preparation method of the gel is the same as that of Example 1.

[0110] Experimental Example 1 Antibacterial test of the composition

[0111] The test sample was the composition prepared in Example 1-4, and the test strains were the fifth generation of Escherichia coli (ATCC25922), the fifth generation of Staphylococcus aureus (ATCC6538), and the fifth generation of Candida albicans (ATCC10231), which were prepared into a bacterial suspension using PBS.

[0112] Testing method: The experiment was carried out with reference to Appendix C4 of GB15979-2002 “Hygiene Standard for Disposable Sanitary Products”. The test was repeated 3 times with the exposure time being 2 minutes, 5 minutes, 10 minutes and 20 minutes respectively.

[0113] The test results are shown in the following table:

[0114] Table 1 Escherichia coli inhibition rate of Examples 1-4

[0115]

[0116] Table 2 Antibacterial rate of Staphylococcus aureus in Examples 1-4

[0117]

[0118] Table 3 Candida albicans inhibition rate of Examples 1-4

[0119]

[0120] It can be seen from the above table that the composition of the present application has a good antibacterial effect, and has a high antibacterial rate for Escherichia coli, Staphylococcus aureus and Candida albicans.

[0121] Experimental Example 2 Effects of the Composition on H8 Cells and Hela Cells

[0122] Experimental methods:

[0123] 1. Cell lines

[0124] H8 cell line (HPV16-positive human cervical cancer epithelial cell line);

[0125] Hela cell line (HPV18 positive human cervical cancer cell line).

[0126] 2. Cell Culture

[0127] H8 cells were cultured in DMEM medium containing 2% fetal bovine serum at 37°C and 5% CO 2 Culture in a saturated humidity constant temperature incubator.

[0128] HeLa cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum at 37°C and 5% CO 2 Culture in a saturated humidity constant temperature incubator.

[0129] 3. Cell Viability Detection

[0130] The cell proliferation inhibition rate was detected by MTT method, and the cell proliferation detection kit was purchased from Shanghai Shangbao Biotechnology Co., Ltd.

[0131] H8 cells and Hela cells in the logarithmic growth phase were prepared into single cell suspensions and the cell density was adjusted to 2×10 5 200 μL per well was inoculated into a 96-well culture plate and incubated at 37°C with 5% CO 2 Culture in an incubator for 4 h.

[0132] The control group was added with serum-free culture medium, the Example 1-4 groups were respectively the compositions prepared in Examples 1-4, the Comparative Example 1-5 groups were respectively the compositions prepared in Comparative Example 1-5, the addition amount was 5 μL, and after 72 h of culture, 20 μL of 5 g / L MTT was added, incubated at 37° C. for 4 h, the supernatant was discarded, 200 μL of DMSO was added to each well, each group was repeated 3 times, the absorbance value at a wavelength of 490 nm was detected, and the cell growth inhibition rate was calculated.

[0133] Cell growth inhibition rate = (1-sample value / control group value) × 100%.

[0134] 4. Expression of HPV16 E6 mRNA and HPV18 E7 mRNA

[0135] Cells from each group were collected and total RNA and Oligo(dT) were extracted from the cells. 15 The expression levels of HPV16 E6 mRNA and HPV18 E7 mRNA were determined by RT-PCR using the primers for reverse transcription, cDNA as template, E6 and E7 as target genes, and β-actin as internal reference gene.

[0136] 5. Experimental Results

[0137] Table 4 Inhibition rate of the composition on H8 cell proliferation

[0138]

[0139] Table 5 Inhibition rate of Hela cell proliferation by the composition

[0140]

[0141] Table 6 Effects of the composition on the expression of HPV16 E6 mRNA and HPV18 E7 mRNA

[0142]

[0143] Experimental Example 3

[0144] 1. Case inclusion criteria

[0145] (1) Patients who meet the Western medical diagnosis criteria of normal vaginal discharge examination, TCT showing squamous intraepithelial lesion or below, high-risk cervical HPV infection, and colposcopy and cervical tissue biopsy showing CINI grade or below;

[0146] (2) Women aged 20-55 years who meet the TCM diagnostic criteria and have a history of sexual activity;

[0147] (3) Regular menstrual cycle and no other factors affecting clinical treatment;

[0148] (4) Voluntarily accept treatment, complete the course of treatment as required, and be able to follow up regularly;

[0149] (5) No history of drug allergy;

[0150] (6) No history of abnormal liver and kidney function, cardiopulmonary insufficiency, or immune system disease;

[0151] (7) The enrolled patients had not received similar drug treatment within six months.

[0152] 2. Case Exclusion Criteria

[0153] (1) Pregnancy during treatment;

[0154] (2) Failure to complete the course of treatment as required or failure to administer medication as prescribed.

[0155] 3. Criteria for determining efficacy

[0156] Recovery: clinical symptoms and signs score ratio ≥ 90%;

[0157] Markedly effective: 70% ≤ clinical symptoms and signs score ratio < 90%;

[0158] Effective: 30%≤clinical symptom and sign score ratio<70%;

[0159] Ineffective: clinical symptoms and signs score ratio <30%.

[0160] Points ratio = (total points before treatment - total points after treatment) / total points before treatment × 100%.

[0161] Total effective rate = effective rate + marked effective rate + cure rate.

[0162] 4. Treatment

[0163] There were 30 patients in the treatment group and 6 patients in the control group. The medication was started on the second day after the menstruation was over. The treatment group was given the gel prepared in Example 1, and the control group was given 5% imiquimod, which was administered once every other day before going to bed. The medication cycle was 6 months. There was no statistically significant difference in age and course of disease between the treatment group and the control group.

[0164] 5. Experimental Results

[0165] Table 7 Clinical efficacy

[0166]

[0167] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A composition, characterized in that The composition is prepared from a Chinese medicine composition and fibronectin. The Chinese medicine composition is prepared from the following components by weight: 6-9 parts of Curcuma zedoaria, 1-3 parts of Artemisia capillaris, 12-14 parts of Astragalus, 7-9 parts of Kochia scoparia, 9-11 parts of Patrinia scabra, 2-4 parts of Sophora flavescens, 4-6 parts of Mentha, 2-3 parts of Lithospermum officinale, 10-12 parts of Viola yedoensis, 4-6 parts of Avena sativa, 12-16 parts of Cnidium monnieri, 5-8 parts of Houttuynia cordata, 10-14 parts of Red Peony Root, 5-8 parts of Ginseng, 6-8 parts of Licorice, 8-10 parts of Portulaca oleracea, 0.5-1.5 parts of Pelargonium, 2-3 parts of Honeysuckle, 5-8 parts of Gentiana scabra, 1-3 parts of Dictamni Cortex, 8-12 parts of Coptis chinensis, 1-3 parts of Scutellaria baicalensis and 5-7 parts of Aquilaria sinensis; and the composition contains 2-4 parts of fibronectin.

2. The composition according to claim 1, characterized in that The Chinese medicine composition is prepared from the following components by weight: 8 parts of Curcuma, 2 parts of Artemisia capillaris, 13 parts of Astragalus, 8 parts of Kochia scoparia, 10 parts of Patrinia, 3 parts of Sophora flavescens, 5 parts of Mentha, 2.5 parts of Lithospermum officinale, 11 parts of Viola yedoensis, 5 parts of Avena sativa, 14 parts of Cnidium monnieri, 7 parts of Houttuynia cordata, 12 parts of Red Peony Root, 6 parts of Ginseng, 7 parts of Licorice, 9 parts of Portulaca oleracea, 1 part of Pelargonium, 2 parts of Honeysuckle, 6 parts of Gentiana scabra, 2 parts of Dictamni, 10 parts of Coptis chinensis, 2 parts of Scutellaria baicalensis and 6 parts of Aquilaria sinensis.

3. The composition according to any one of claims 1 to 2, characterized in that The dosage form of the composition is drops, tincture, powder, ointment, powder, emulsion, gel and suppository.

4. The composition according to claim 3, characterized in that The dosage form of the composition is a gel.

5. The method for preparing the composition according to any one of claims 1 to 4, characterized in that: The following steps are involved: (1) Take parts by weight of Curcuma zedoaria, Astragalus, Sophora flavescens, Lithospermum officinale, Viola yedoensis, Cnidium monnieri, Houttuynia cordata, Coptis chinensis, and Aquilaria sinensis, grind them, add ethanol to perform reflux extraction, filter and concentrate, recover ethanol, and obtain a concentrated solution 1; (2) taking parts by weight of Artemisia capillaris, Kochia scoparia, Patrinia scabra, Mentha, Oat kernel, Red Peony Root, Ginseng, Licorice, Portulaca oleracea, Geranium, Honeysuckle, Gentiana scabra, Dictamni Cortex, and Scutellaria baicalensis, adding water for decoction to obtain a decoction, and concentrating to obtain a concentrate 2; (3) The concentrated solution 1, the concentrated solution 2 and the fibronectin are mixed to obtain a composition.

6. The preparation method according to claim 5, characterized in that: The amount of ethanol added in step (1) is 8-12 times the mass of the traditional Chinese medicine.

7. The preparation method according to claim 5, characterized in that: The volume fraction of ethanol in step (1) is 70-90%.

8. The preparation method according to claim 5, characterized in that: The number of reflux extractions in step (1) is 1-3 times, and the time for each reflux extraction is 1-3 hours.

9. The preparation method according to claim 5, characterized in that: The amount of water added in step (2) is 8-14 times the mass of the Chinese medicine.

10. The preparation method according to claim 5, characterized in that: The number of decoctions in step (2) is 1-3 times, and the decoction time each time is 2-4 hours.

11. Use of the composition according to any one of claims 1 to 4 in the preparation of a medicament for preventing or treating HPV infection.

12. The use according to claim 11, characterized in that: The medicine is used for preventing or treating vaginitis, cervical erosion and cervical cancer caused by HPV infection.

13. Use of the composition according to any one of claims 1 to 4 in the preparation of antibacterial products, characterized in that: The product is used to inhibit Escherichia coli, Staphylococcus aureus or Candida albicans.

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