CN112022800B

CN112022800B - Composition, vaginal gel prepared from composition and application of vaginal gel in resisting human papilloma virus

Info

Publication number: CN112022800B
Application number: CN202010992467.4A
Authority: CN
Inventors: 陆启东; 李靖; 付修远; 孙雨薇
Original assignee: Nanjing Aurora Biotechnology Co ltd
Current assignee: Nanjing Aurora Biotechnology Co ltd
Priority date: 2020-09-21
Filing date: 2020-09-21
Publication date: 2023-06-09
Anticipated expiration: 2040-09-21
Also published as: CN112022800A

Abstract

The invention discloses a composition, a vaginal gel prepared from the composition and application of the vaginal gel in resisting human papilloma virus, wherein the gel functional components comprise beta-glucan and hyaluronic acid, the beta-glucan and the hyaluronic acid are carriers of gel products and the gel products are effective components for resisting human papilloma virus, and the vaginal gel has good biocompatibility and almost no toxic or side effect. The beta-glucan and hyaluronic acid have synergistic effect of resisting human papillomavirus, and have the advantages of stability, moderate viscosity, comfortable use and no influence on vaginal bacteria.

Description

Composition, vaginal gel prepared from composition and application of vaginal gel in resisting human papilloma virus

Technical Field

The invention relates to the technical field of gynecological care products, in particular to a product of vaginal gel for resisting human papilloma virus.

Background

High-risk HPV infection is easy to cause cervical intraepithelial neoplasia, cervical carcinoma and the like, and effective intervention in the early stage of HPV infection is significant for improving prognosis. Among them, long-term infection with HPV16, HPV18 subtypes has been demonstrated as a major cause of cervical cancer. There is no specific prevention and treatment drug for HPV on the market at present, and HPV vaccines are mainly suitable for women without sexual activity. Therefore, the development of novel anti-HPV health products and medicines has important significance. The drug for treating cervical HPV infection commonly uses interferon such as Xin Funing (recombinant human interferon alpha 2b vaginal effervescent capsule) to inhibit the replication and transcription of viral nucleic acid by inducing the production of enzymatically active antiviral protein in target cells in theory, but the efficacy and toxic and side effects are still controversial. More than 80 types of HPV have been fully identified, HPV being a double-stranded DNA virus comprising a number of different subtypes, and being classified into low-risk and high-risk types. The clinical lesions of HPVs infected with different genotypes are also different, and when high-risk HPV viruses are infected, 80% of cervical cancers which eventually develop into cervical cancers are caused by type 4 HPV infection of types 16, 18, 3l and 45, especially type 16/18 if not treated in time. Although vaccines against HPV have been developed, their safety and their impact on the immune system remain to be considered. I.e. HPV vaccines are currently still far from maturation and start from large-scale and large-scale popularization.

US20190343870A1 discloses an application of alginic acid sulfate in preparing medicines and health products for preventing and treating diseases caused by human papilloma virus, and experiments prove that alginic acid sulfate has strong inhibition effect on HPV infection.

The use of PsV to nonspecifically pack DNA was reported in Journal of Virology,79 (5) 2839-2846, 2005, in which plasmids of the HPVL1 and L2 genes were co-transfected with a reporter plasmid to construct a pseudo-viral particle encapsulating the reporter plasmid. The HPV pseudovirus system is widely used for screening and identifying HPV resisting products at present.

Beta-glucan is a kind of macromolecular polysaccharide widely existing in microorganisms, plants and animals, the main chain structure is connected by beta-1, 3-glycosidic bond, and beta-1, 3-glucan is an active substance with immunostimulation effect. The beta-1, 3-glucan has the functions of resisting cancer, bacteria, viruses, fungi, parasites, cholesterol and blood fat, promoting wound healing and the like. The preparation of anti-HPV dressings using beta-glucan has been reported, CN110478514a discloses the preparation of anti-HPV virus dressings using sulfated yeast glucan as an active ingredient, and the addition of phytic acid ketone, flavone, silicone for combined use. EP3116519B1 discloses a topical composition comprising coriolus versicolor extract for preventing and/or treating vaginal or cervical diseases caused by infectious agents by vaginal or cervical administration, the composition having the effect of enhancing immunity of genital anti-infective agents and re-epithelizing damaged tissues, the main active compounds in coriolus versicolor extract being β -glucan, polysaccharide peptide Krestin (PSK) and polysaccharide peptide (PSP); carboxymethyl-beta-glucan may be added. The GiadaLavitola et al published paper studied the effect of using vaginal administration and carboxymethyl-Beta-Glucan on vaginal flora and cervical epithelial tissue, and the results of 6 months of observation experiments showed that carboxymethyl-Beta-Glucan can comprehensively improve vaginal health and reduce the risk of uterine tumors (Effects on Vaginal Microbiota Restoration and Cervical Epithelialization in Positive HPV Patients Undergoing Vaginal Treatment with Carboxy-Methyl-Beta-Glucan).

The prior hyaluronic acid (also called hyaluronic acid, sodium hyaluronate and sodium hyaluronate) and derivatives thereof vaginal gel are mainly used for vaginal dryness supplementary treatment and promote the natural recovery of small damage caused by friction in vaginal mucosa. US20190351013A1 discloses an arginine-rich polypeptide composition and methods of use thereof, the arginine-rich polypeptide having at least 9 arginine residues that comprise at least 10% of the amino acid residues in the polypeptide, the arginine-rich polypeptide being useful in methods of inhibiting binding of Human Papillomavirus (HPV) to cells, inhibiting intracellular processing of Human Papillomavirus (HPV), and optionally inhibiting Human Papillomavirus (HPV) using a complex of the arginine-rich polypeptide and hyaluronic acid.

The anti-HPV products containing beta-glucan in the prior art have insufficient effect on HPV removal; on the basis of beta-glucan, hormones, interferons (such as recombinant human interferon alpha-2 b), extracts or chemical synthetic drugs and the like are added, so that on one hand, potential safety hazards and side effects are caused, beneficial flora in the vagina is influenced, on the other hand, for gel products containing beta-glucan and resisting HPV, the added substances are unfavorable for the formation of gel dosage forms, the physicochemical properties of the additives need to be considered, and the stability is relatively poor; making the process of preparing gel dosage forms relatively complex.

Disclosure of Invention

The present invention aims to address the deficiencies of the prior art by providing a composition comprising beta-glucans and hyaluronic acid, the beta-glucans comprising a mixture of one or more of beta-glucans, derivatives of beta-glucans; the hyaluronic acid comprises hyaluronic acid, and one or more of hyaluronic acid derivatives.

Further, 1-10 parts of beta-glucan and 1-10 parts of hyaluronic acid; preferably, 3-9 parts of beta-glucan and 2-7 parts of hyaluronic acid; more preferably, 5-6 parts of beta-glucan and 4-5 parts of hyaluronic acid; the gel containing β -glucan and hyaluronic acid in this ratio exhibits excellent HPV-inhibiting and preventing effects.

Further, in the beta-glucan or the derivative of beta-glucan, the ratio of beta-1, 3 glycosidic bond to beta-1, 6 glycosidic bond is 1:99 to 99:1, preferably, the ratio is 1:9 to 9:1, more preferably, (7-9): 3-1.

Further, in the beta-glucan or the derivative of beta-glucan, the ratio of glucan having a main chain of beta-1, 3 glycosidic bond to glucan having a main chain of beta-1, 6 glycosidic bond is 1:99 to 99:1, preferably, the ratio is 1:9 to 9:1, more preferably, (7-9): 3-1.

On the human immunity level, the beta-glucan improves the activity of corresponding T lymphocytes, macrophages, dendritic cells and other natural immune system cells, and promotes the differentiation and activation of the T lymphocytes; hyaluronic acid contributes to the activation of cells of the immune system and also to the binding and phagocytosis of immune cells to cells infected with HPV. Whereas glucans having 1,3 glycosidic linkages as the main chain, glucans having 1,6 side chains have a greater activity in activating immune responses.

Further, the content of beta-glucan is 1-100mg/g, preferably 5-25mg/g, more preferably 18mg/g, and the content of hyaluronic acid is 1-100mg/g, preferably 5-25mg/g, more preferably 12mg/g;

further, the beta-glucan class average molecular weight is 50,000 to 3,000,000, preferably 100,000 to 1,000,000; the average molecular weight of the hyaluronic acid is 500,000 to 3,000,000, preferably 750,000 to 1,000,000.

In the concentration and molecular weight range, the adhesive force is kept better, the vaginal gel is less or even avoided being left, and bad body feeling and pollution to clothes and bed clothes are brought; can continuously exert the gel effect and increase the contact time of the gel and the vaginal cells. In addition, in this concentration and molecular weight range, the gel exhibits excellent HPV-inhibiting and preventing effects.

Further, derivatives of beta-glucan include carboxymethylated beta-glucan, sulfated glucan salts; derivatives of hyaluronic acid include hyaluronate.

The invention also provides a gel which comprises beta-glucan, hyaluronic acid and physiologically acceptable auxiliary materials.

Further, the gel is a vaginal gel.

In the concentration and molecular weight range, the adhesive force is kept better, the vaginal gel is less or even avoided being left, and bad body feeling and pollution to clothes and bed clothes are brought; can continuously exert the gel effect and increase the contact time of the gel and the vaginal cells.

In addition, in this concentration and molecular weight range, the gel exhibits excellent HPV-inhibiting and preventing effects.

Further, the physiologically acceptable auxiliary materials are one or a mixture of several of pure water, chitosan, carbomer, preservative, osmotic pressure regulator, pH regulator, prebiotics, probiotics and glycerin.

The pH regulator is preferably a phosphate buffer system; the osmolality regulator is preferably sodium salt, the preservative is preferably nipagin and derivatives thereof, and the preservative inhibits the growth of microorganisms, in particular bacteria and fungi.

The invention further provides application of the composition in preparing HPV products, wherein the composition is beta-glucan and hyaluronic acid, and the beta-glucan comprises beta-glucan and one or more of derivatives of the beta-glucan; the hyaluronic acid comprises hyaluronic acid, and one or more of hyaluronic acid derivatives.

Further, the product is a vaginal gel.

Further, the daily amount of the product is 1 to 10g, preferably 3 to 5g.

A method of using a vaginal gel, characterized in that the vaginal gel prepared with the composition of any one of claims 1-3, or the gel of any one of claims 4-7, is delivered to the vagina.

Further, the amount delivered is 3-5g per delivery.

Further, delivery is performed using a delivery instrument.

The vaginal gel provided by the invention has no negative effect on vaginal flora, and is beneficial to the vaginal flora even under the condition of not adding prebiotics; such as lactobacilli in the vagina.

On the immune level of the organism, the beta-glucan can activate macrophages, neutrophils and the like, so that the content of the leucocyte, the cytokinin and the special antibody can be improved, and the immune system of the organism can be comprehensively stimulated. The beta-glucan can quickly restore the ability of lymphocytes of injured organisms to produce cytokine (IL-1) and effectively regulate the immune function of the organisms. Beta-glucan can promote IgM antibody production in vivo to enhance humoral immunity. Hyaluronic acid has an activating effect on immune cells.

The glucan of the invention is obtained from plant sources or from biotechnological processes, including synthesis via bacteria, fungi or enzymes. Suitably, the hyaluronic acid of the invention is obtained from fermentation or enzymatic synthesis. The glucan of the present invention preferably comprises yeast-produced beta-glucan.

The hyaluronic acid of the invention is obtained from animal sources or from biotechnological processes, including via bacterial or enzymatic synthesis. Suitably, the hyaluronic acid of the invention is obtained from fermentation or enzymatic synthesis. The hyaluronic acid of the invention preferably contains hyaluronic acid produced by the microorganism bacillus subtilis.

The gel product has simpler production process and stable gel property. The production method of the gel product comprises the steps of respectively preparing beta-glucan gel, preparing hyaluronic acid gel, mixing, adding or not adding other components, and packaging the finished product; wherein the mixing and the adding are not sequential.

The invention also provides a method for producing the gel product, which comprises the steps of mixing the beta-glucan raw material and the hyaluronic acid raw material, adding or not adding other components in the mixing step to prepare the gel, adding or not adding other components, and packaging the finished product.

Beta-glucan and hyaluronic acid are both carriers of gel products and anti-HPV efficacy components of the gel products, and have good biocompatibility and almost no toxic or side effect. No other excipient, carrier or matrix is needed to be added, and the gel effect is shown. The anti-HPV can be obtained without adding hormone, interferon, extract or chemical synthesis medicine.

The above description of products containing beta-glucans and hyaluronic acids, in particular gel products, does not exclude other excipients of the gel products and does not exclude hormones, interferons, extracts or chemically synthesized drugs. In fact, the gel containing β -glucan and hyaluronic acid in the present application may be added with other substrates, carriers, excipients, and the like, for example, chitosan, carbomer, and the like. It is also possible to add interferon, hormone, extract or chemical synthesis medicine, etc., without affecting the activity of interferon, hormone, extract or chemical synthesis medicine. In particular, plant extracts having antiviral effects are used without negatively affecting the activity of the plant extracts against HPV.

In one embodiment, the main active ingredients of the vaginal gel of the present invention are beta-glucans and hyaluronic acids. In another embodiment, the only active ingredients of the vaginal gel of the present invention are beta-glucans and hyaluronic acid; in particular embodiments, the amount of other anti-HPV active ingredient is less than 1%, preferably less than 0.5%, 0.3%, 0.1%, 0.05%, or 0.01%.

The beneficial effects of the invention include: (1) Beta-glucan and hyaluronic acid are not only carriers of gel products, but also anti-HPV efficacy components of the gel products, and have good biocompatibility and almost no toxic or side effect; (2) On the immune level of the organism, beta-glucan and hyaluronic acid can activate the immune cell response of HPV; (3) The beta-glucan and hyaluronic acid have synergistic inhibition effect on the adsorption and immersion processes of HPV viruses; (4) The product is stable, the viscosity is moderate, the use is comfortable, and the subjective feeling is good; (5) no inhibition to vaginal bacteria.

The term "administration" as used herein is vaginal or cervical administration, directly to the genital area, in particular the vulva, vagina or cervix.

As used herein, "anti" includes prophylactic, inhibitory, and scavenging effects.

As used herein, "extract" generally refers to a composition containing biologically active chemical components and/or compounds isolated from animals, plants, microorganisms.

As used herein, "beta-glucan" is a type of macromolecular polysaccharide, and refers to a type of polysaccharide consisting of monosaccharides of glucose, wherein glucose units are connected by glycosidic bonds, and glucose is connected by beta-1, 3-glycosidic bonds, beta-1, 6-glycosidic bonds, beta-1, 4-glycosidic bonds.

As used herein, "hyaluronic acid", also known as hyaluronic acid, is a mucopolysaccharide formed by alternately connecting glucuronic acid and N-acetylglucosamine as disaccharide units; the common form on the market is sodium hyaluronate or sodium hyaluronate.

As used herein, "average molecular weight" is the molecular weight of a commercially available product; or mixing and averaging similar products with different molecular weights.

Detailed Description

The invention will be further illustrated by the following examples, which are not intended to limit the scope of the invention, in order to facilitate the understanding of those skilled in the art.

Example 1

A gel contains beta-glucan and hyaluronic acid, 6 parts of beta-glucan and 4 parts of hyaluronic acid.

As test gel against HPV effects: the beta-glucan is prepared from beta-glucan and hyaluronic acid, wherein the content of the beta-glucan is 18mg/g, and the content of the hyaluronic acid is 12mg/g; the beta-glucan has an average molecular weight of 100,000 to 1,000,000, about 500,000; hyaluronic acid has an average molecular weight of 750,000 to 1,000,000, about 800,000.

HeLa cells are cancer cells transformed from normal cervical cells, and as an infecting object of HPV pseudoviruses in the present invention, a suspension of HeLa cells is prepared in a conventional manner; HPV16 pseudovirus particles were prepared in a conventional manner; anti-HPV gel was diluted 10-fold as original dilution solution of anti-HPV gel, and inhibition assay of HPV viral activity by anti-HPV virus.

(1) 100 μl of the cell suspension was added to a 96-well microplate to achieve a cell volume of one hundred thousand/ml. For each concentration and control, 5 wells were made and the relevant data averaged over 5 sets of values; 100. Mu.l of growth solution+100. Mu.l of cell suspension as a cell growth blank; (2) 50. Mu.l HPV16 pseudovirion liquid+50. Mu.l growth liquid+100. Mu.l cell suspension as positive control; (3) Continuously diluting the original dilution solution of the anti-HPV gel by 5 times, 10 times, 25 times and 100 times by using growth solution, wherein 50 mu.l of HPV16 pseudovirus particle solution is added with 50 mu.l of the dilution solution of the anti-HPV gel and 100 mu.l of cell suspension; (4) Beta-glucan gel with the content of 18mg/g and the average molecular weight of 100,000 to 1,000,000 is firstly diluted 10 times, then the solution is taken and diluted 5 times, 50 mu.l of HPV16 pseudovirus particle solution, 50 mu.l of gel dilution solution and 100 mu.l of cell suspension are taken as a comparison example 1 beta-glucan control group; (5) Hyaluronic acid gel having an average molecular weight of 1,000,000 to 2,000,00 at a concentration of 12mg/g was diluted 10-fold, then the solution was taken and diluted 5-fold again, and 50. Mu.l of HPV16 pseudovirion liquid + 50. Mu.l of gel diluted solution + 100. Mu.l of cell suspension was used as the hyaluronic acid control group of comparative example 2.

96-well microplates were incubated at 37℃with 5% CO ₂ Incubate for 24 hours, then discard the culture medium and wash several times with PBS. And (5) observing under a fluorescence microscope, and comparing the cell fluorescence values of each group with those of a control group.

TABLE 1 inhibition of HPV pseudoviruses by gel

As can be seen from Table 1, the gel diluted 1000 times, i.e., the diluted solution of hyaluronic acid at a concentration of 18. Mu.g/g and hyaluronic acid at a concentration of 12. Mu.g/g, still exhibited an inhibitory effect; the gel dilutes the solution by 10 times, and the inhibition effect is very obvious; almost no fluorescence; solutions of beta-glucan and hyaluronic acid exhibit concentration dependence, and particularly in 10-fold, 50-fold and 100-fold dilutions, the inhibition effect is relatively remarkable. When the dilution factor is too large, for example, 1000-fold or 250-fold, the inhibition effect is relatively weak.

In addition, comparing the gel 50-fold diluent group, the beta-glucan control group and the hyaluronic acid control group can be seen to have a synergistic effect on inhibiting HPV, and the combined use shows obvious inhibition effect compared with the beta-glucan control group and the hyaluronic acid control group.

Beta-glucan is a common anti-HPV material capable of inhibiting HPV, and therefore is a common dressing for HPV gels and suppositories. While the applicant is still under investigation for synergistic mechanisms, possible mechanisms of action are: on the viral replication level, viruses are divided into adsorption, invasion, uncoating, biosynthesis, assembly and release processes; the beta-glucan has adsorption effect on viruses, the hyaluronic acid is combined with cell receptors, the adsorption process of the viruses and the combination process of the hyaluronic acid and the cell receptors are also inhibited, the beta-glucan and the hyaluronic acid are both macromolecular polysaccharides, and the chain hydrophilic hydroxyl groups are mutually wrapped to form skeleton polymerization, so that a protective layer which can adsorb HPV viruses and inhibit the combination of the HPV and the cell receptors is formed on the surface of the cells; the combined use of beta-glucan and hyaluronic acid thus greatly reduces the likelihood of adsorptive contact of the virus with the cellular receptor.

Example 2

A gel comprises 5 parts of beta-glucan and hyaluronic acid, and 5 parts of hyaluronic acid.

Example 3

A gel comprises beta-glucan and hyaluronic acid, wherein the beta-glucan comprises 3 parts of beta-glucan and 7 parts of hyaluronic acid.

Example 4

A gel comprises beta-glucan and hyaluronic acid, wherein the beta-glucan comprises 8 parts of beta-glucan and 2 parts of hyaluronic acid.

Example 5

A gel comprises beta-glucan and hyaluronic acid, wherein the beta-glucan comprises 1 part and 10 parts of hyaluronic acid.

Example 6

A gel comprises beta-glucan and hyaluronic acid, 10 parts of beta-glucan and 1 part of hyaluronic acid.

Example 7

A gel comprising beta-glucan and hyaluronic acid, the beta-glucan having an average molecular weight of about 50,000; the average molecular weight of hyaluronic acid is about 2,000,000.

Example 8

A gel comprises beta-glucan having an average molecular weight of about 3,000,000 and hyaluronic acid having an average molecular weight of about 1,000,000.

Example 9

A gel comprising beta-glucan and hyaluronic acid, the beta-glucan having an average molecular weight of about 50,000; the average molecular weight of hyaluronic acid is about 500,000.

Example 10

A gel comprising beta-glucan and hyaluronic acid, the beta-glucan having an average molecular weight of about 500,000; the average molecular weight of hyaluronic acid is about 750,000.

Example 11

A gel comprising beta-glucan and hyaluronic acid, the beta-glucan having an average molecular weight of about 1,000,000; the average molecular weight of hyaluronic acid is about 1,000,000.

Comparative example 1

A gel comprising beta-glucan and hyaluronic acid, the beta-glucan having an average molecular weight of about 10,000; the average molecular weight of hyaluronic acid is about 100,000.

Comparative example 2

A gel comprising beta-glucan and hyaluronic acid, the beta-glucan having an average molecular weight of about 10,000; the average molecular weight of hyaluronic acid is about 50,000.

Comparative example 3

A gel comprising beta-glucan and hyaluronic acid, the beta-glucan having an average molecular weight of about 30,000; the average molecular weight of hyaluronic acid is about 100,000.

Comparative example 4

A gel comprising beta-glucan and hyaluronic acid, the beta-glucan having an average molecular weight of about 40,000; the average molecular weight of hyaluronic acid is about 300,000.

Test gel as a continuous effect of viscosity test:

a gel comprising beta-glucan and hyaluronic acid; 6 parts of beta-glucan and 4 parts of hyaluronic acid; the content of beta-glucan is 18mg/g, and the content of hyaluronic acid is 12mg/g.

The gels were tested for viscosity at 50rad/s after 1 hour in a 37℃water bath, the rheological properties were tested using a frequency sweep shear test, each set repeated 5 times, and the data averaged.

TABLE 2 influence of molecular weight of beta-glucan and hyaluronic acid on gel viscosity

Sequence number	Group of	Viscosity Pa.s
			1	Example 7	0.897
2	Example 8	0.168
			3	Example 9	0.063
4	Example 10	0.102
			5	Example 11	0.126
6	Comparative example 1	0.028
			7	Comparative example 2	0.019
8	Comparative example 3	0.041
			9	Comparative example 4	0.048

As can be seen from table 2, the medium and high molecular weight β -glucan and hyaluronic acid can maintain the viscosity of the gel, less leakage of the gel, and maintain an appropriate degree of lubricity; on the other hand, the smaller the molecular mass of the hyaluronic acid is, the stronger the permeability is, and the immune efficacy and the HPV effect are better exerted in the body.

Example 12

A gel contains beta-glucan and hyaluronic acid, wherein the content of beta-glucan is 1mg/g, and the content of hyaluronic acid is 100mg/g.

Example 13

A gel contains beta-glucan and hyaluronic acid, wherein the content of beta-glucan is 100mg/g, and the content of hyaluronic acid is 1mg/g.

Example 14

A gel contains beta-glucan and hyaluronic acid, wherein the content of beta-glucan is 5mg/g, and the content of hyaluronic acid is 25mg/g.

Example 15

A gel contains beta-glucan and hyaluronic acid, wherein the content of beta-glucan is 25mg/g, and the content of hyaluronic acid is 5mg/g.

Example 16

A gel contains beta-glucan and hyaluronic acid, wherein the content of beta-glucan is 18mg/g, and the content of hyaluronic acid is 12mg/g.

Example 17

A gel contains beta-glucan and hyaluronic acid, wherein the content of beta-glucan is 1mg/g, and the content of hyaluronic acid is 1mg/g.

Example 18

A gel contains beta-glucan and hyaluronic acid, wherein the content of beta-glucan is 100mg/g, and the content of hyaluronic acid is 100mg/g.

As a test gel for content: the beta-glucan is prepared from 6 parts of beta-glucan and 4 parts of hyaluronic acid; the beta-glucan has an average molecular weight of 100,000 to 1,000,000, about 500,000; hyaluronic acid has an average molecular weight of 750,000 to 1,000,000, about 800,000.

TABLE 3 influence of the content of beta-glucan and hyaluronic acid on the gel viscosity

As can be seen from Table 3, the content also directly affects the viscosity of the gel, which exhibits a positive correlation with the content. The content of beta-glucan and hyaluronic acid in the gel is based on the proportion relation, HPV resistance effect and daily use amount of consumers, and the dissolution condition and production efficiency of the beta-glucan and the hyaluronic acid in the production process are considered.

In the case of high amounts of both beta-glucan and hyaluronic acid, for example, 100mg/g beta-glucan and 100mg/g hyaluronic acid, it is almost impossible to form a gel that is changeable and effectively totally soluble, and in particular, it is difficult to form a gel that is subjectively felt by vaginal accommodation. The data variance in the parallel test of example 12 and example 13 is larger than that of the other groups; too high content and poor product stability are also indicated.

Under the condition that the dosage of the beta-glucan and the hyaluronic acid is relatively low, for example, the content of the beta-glucan is 1mg/g, the content of the hyaluronic acid is 1mg/g, and the product is in a solution state and is in a non-gel state. The subject matter of the gel product is defined, in fact, by the content of beta-glucan and the content of hyaluronic acid.

Beta-glucan content is 5-25mg/g, hyaluronic acid content is 5-25mg/g, dissolution and production are relatively efficient, and gel formation and viscosity maintenance are very good. Furthermore, daily doses of 1-10g, preferably 3-5g, are very suitable for vaginal gel delivery and vaginal accommodation and subjective perception by humans.

Example 19

A gel comprises sulfated beta-glucan and hyaluronic acid, wherein the beta-glucan content is 18mg/g, and the hyaluronic acid content is 12mg/g.

Example 20

A gel comprises carboxymethylated beta-glucan and hyaluronic acid, wherein the beta-glucan content is 18mg/g, and the hyaluronic acid content is 12mg/g.

The method of example 1 was used to test the selection of sulfated and carboxymethylated beta-glucans as components of the beta-glucans, respectively, and to determine the anti-HPV effect on the product. The gels were diluted 100-fold.

TABLE 4 inhibition of HPV pseudoviruses by gel

As can be seen from table 4, both the sulfated and carboxymethylated β -glucan groups showed a stronger resistance against HPV than the β -glucan group, in particular sulfated β -glucan. The water solubility of the polysaccharide is enhanced through sulfation, on the other hand, the sulfated polysaccharide not only shows stronger HPV resistance, but also contains the HPV resistance effect shown by sulfated natural polysaccharide such as carrageenan. And carboxymethylated beta-glucan also shows better anti-HPV effect.

Example 21

A gel comprises beta-glucan and hyaluronic acid, wherein the content of beta-glucan is 18mg/g, the content of hyaluronic acid is 12mg/g, and the gel further comprises 5mg/g of chitosan, 3mg/g of carbomer, 0.1mg/g of nipagin and 10mg/g of sodium chloride.

Example 22

A gel contains beta-glucan and hyaluronic acid, wherein the beta-glucan content is 18mg/g, the hyaluronic acid content is 12mg/g, and the gel further comprises oligosaccharide 2mg/g and glycerin 20mg/g.

The vaginal gel product of the invention is stable, chitosan, carbomer, preservative, osmotic pressure regulator, prebiotics, glycerin, and the like, and common auxiliary materials in the gel can be applied to the gel in the invention.

Example 23

A method of using a vaginal gel using the gel of example 1, the gel product was delivered to the vagina using a delivery device, the daily amount of product was 3g.

It will be understood by those skilled in the art that all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs unless defined otherwise. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the prior art and will not be interpreted in an idealized or overly formal sense unless expressly so defined herein.

It should be understood that the detailed description of the technical solution of the present invention, given by way of preferred embodiments, is illustrative and not restrictive. Modifications of the technical solutions described in the embodiments or equivalent substitutions of some technical features thereof may be performed by those skilled in the art on the basis of the present description; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A composition for preventing or inhibiting human papillomavirus, characterized by consisting of 1-10 parts of beta-glucan or a derivative thereof having an average molecular weight of 100,000 to 1,000,000 and 1-10 parts of hyaluronic acid or a derivative thereof having an average molecular weight of 500,000 to 3,000,000, wherein the beta-glucan derivative is carboxymethylated beta-glucan or sulfated beta-glucan, and the hyaluronic acid derivative is hyaluronate.

2. The composition of claim 1, wherein the ratio of β -1,3 glycosidic linkages to β -1,6 glycosidic linkages in β -glucan or a derivative thereof is from 1:99 to 99:1; or the ratio of glucan with beta-1, 3 glycosidic bond in the main chain to glucan with beta-1, 6 glycosidic bond in the main chain is 1:99 to 99:1.

3. The composition of claim 2, wherein the ratio of β -1,3 glycosidic linkages to β -1,6 glycosidic linkages in β -glucan or a derivative thereof is from 1:9 to 9:1; or the ratio of glucan with beta-1, 3 glycosidic bond in the main chain to glucan with beta-1, 6 glycosidic bond in the main chain is 1:9 to 9:1.

4. A composition according to claim 3, wherein the ratio of β -1,3 glycosidic linkages to β -1,6 glycosidic linkages in β -glucan or a derivative thereof is (7-9): 3-1; or the ratio of glucan with beta-1, 3 glycosidic bond in the main chain to glucan with beta-1, 6 glycosidic bond in the main chain is (7-9): 3-1.

5. The composition of claim 1, wherein the beta-glucan or derivative thereof is 3-9 parts and the hyaluronic acid or hyaluronate is 2-7 parts.

6. The composition of claim 5, wherein the beta-glucan or derivative thereof is 5-6 parts and the hyaluronic acid or hyaluronate is 4-5 parts.

7. A gel for preventing or inhibiting human papilloma virus, characterized in that it is made of the composition according to any one of claims 1 to 6 and physiologically acceptable excipients, the content of beta-glucan or its derivatives being 1-100mg/g, the content of hyaluronic acid or hyaluronate being 1-100mg/g, the gel being a vaginal gel.

8. The gel of claim 7, wherein the beta-glucan or derivative thereof is present in an amount of 5-25mg/g and the hyaluronic acid or hyaluronate is present in an amount of 5-25mg/g.

9. The gel of claim 8, wherein the beta-glucan or derivative thereof is present in an amount of 18mg/g and the hyaluronic acid or hyaluronate is present in an amount of 12mg/g.

10. The gel according to any one of claims 7-9, wherein the physiologically acceptable auxiliary material is one or a mixture of several of pure water, chitosan, carbomers, preservatives, tonicity modifiers, prebiotics, probiotics, glycerol.

11. Use of a composition according to any one of claims 1-6 for the preparation of a product for preventing or inhibiting human papillomavirus.

12. The use according to claim 11, wherein the product is a vaginal gel; the daily dosage is 1-10g.

13. Use according to claim 12, characterized in that the daily amount is 3-5g.

14. A method of preparing a gel according to any one of claims 7 to 10, wherein: preparing gel of beta-glucan or derivatives thereof and gel of hyaluronic acid or hyaluronate respectively, then mixing, adding or not adding other components, and packaging the finished product, wherein the mixing and the adding of the other components are not carried out sequentially or are carried out simultaneously; or mixing beta-glucan or its derivative with hyaluronic acid or hyaluronate, adding or not adding other components, preparing gel, adding or not adding other components, and packaging to obtain the final product, wherein the mixing and adding other components are not carried out sequentially or simultaneously.