Anti-HPV infection soapberry peptide solution and preparation method thereof

Abstract

The present invention provides use of a Soapberry peptide solution in the preparation of an anti-HPV infection lotion, gel, or suppository. The preparation method for the Soapberry peptide solution comprises the following steps: step S1, preparation of a Soapberry pulp homogenate; step S2, enzymolysis; step S3, ultrafiltration and nanofiltration; and step S4, uniformly mixing a Soapberry peptide liquid with an additional component to obtain the Soapberry peptide solution.

Description

A soapberry peptide solution for resisting HPV infection and its preparation method Technical Field

[0001] The present invention relates to the field of medical and health technology, and in particular to a soapberry peptide solution for resisting HPV infection and a preparation method thereof. Background Art

[0002] HPV, or human papillomavirus, is a spherical DNA virus belonging to the genus Papillomatovirus A of the family Papovaviridae. It can cause proliferation of the squamous epithelium of the human skin and mucous membranes. Symptoms include common warts and genital warts (condyloma acuminata). With the rapid rise in the incidence of genital warts among STDs and the increasing incidence of cervical and anal cancers, HPV infection is attracting increasing attention.

[0003] Currently, treatments for HPV symptoms primarily include vaccines, physical therapy, medications, and immunotherapy. Vaccines have limitations and lack broad-spectrum protection. Furthermore, vaccine supply is limited, and the cost of vaccination is high, failing to meet public demand. Physical therapy primarily targets tumors caused by the virus, rather than eliminating the virus at its source. Immunotherapy is expensive, unstable, and involves complex procedures. Medication remains the most convenient and effective treatment for HPV, but existing effective medications for the virus are limited and have significant side effects.

[0004] my country boasts a rich resource of Traditional Chinese Medicine (TCM), a tradition passed down for thousands of years. It boasts a vast database of applications and enjoys high public recognition and trust. TCM contains a variety of active ingredients with a wide range of pharmacological actions, acting on multiple levels and targets with minimal side effects. This holds great promise for the development of new drugs to treat HPV.

[0005] Soapberry, also known as Soapberry, has very strong medicinal value. Its pericarp, seed, kernel, root, skin, leaf can be used as pharmaceutical raw materials. Modern pharmacological research has proved that Soapberry also has pharmacological effects such as antibacterial, antitumor, anti-stress, antiviral, cardio-liver protection, and immunomodulatory. Simultaneously, Soapberry contains natural surfactant, which contains multiple beneficial substances such as terpenoid steroid saponins, amino acids, proteins, vitamins, oleic acid, grease, etc. However, currently, its pericarp is mainly used to extract soapberry saponins, and its stone is discarded in a large number, causing waste of resources and certain damage to the natural and social environment.

[0006] Currently, peptide products on the market primarily include animal-derived and plant-derived peptides. Among the numerous plant-based raw materials, soapberry peptides have yet to be developed and extracted. Research on soapberry has primarily focused on extracting essential oils and saponins, and most other peptides have a bitter and fishy taste, making them difficult to commercialize. Furthermore, existing technologies for peptide extraction and preparation suffer from issues such as poor enzymatic hydrolysis, complex processes, and high costs, making them unsuitable for industrial production.

[0007] In view of this, the present invention utilizes the less studied soapberry kernel to provide a soapberry peptide solution, which can be used to resist HPV infection and can be made into clinically or pharmaceutically acceptable dosage forms such as lotions, gels or suppositories after adding excipients. Summary of the Invention

[0008] The purpose of the present invention is to overcome the shortcomings of the existing technology and provide an anti-HPV infection soapberry peptide solution and its preparation method that has good anti-HPV infection effect, good antibacterial effect, low toxic and side effects, simple preparation process, and is affordable for ordinary people.

[0009] To achieve the above object, the technical solution adopted by the present invention is: a method for preparing a soapberry peptide solution for resisting HPV infection, comprising the following steps:

[0010] Step S1, preparation of soapberry meal homogenate: taking soapberry meal, adding it to purified water, and processing it in a high-speed tissue crushing homogenizer for 10 to 15 minutes to obtain soapberry meal homogenate;

[0011] Step S2, enzymolysis: the soapberry meal homogenate prepared in step S1 is placed in an enzymolysis tank, stirred at 88-92° C. for 10-15 minutes, and then cooled to 45-55° C. At this temperature, the pH is adjusted to 9-10 using a 0.8-1.2 mol / L NaOH solution, and alkaline protease is added thereto for enzymolysis; the NaOH solution is added during the enzymolysis process to maintain a relatively stable pH; after the enzymolysis is completed, the enzyme is inactivated by heating, the mixture is cooled, discharged, centrifuged, and the supernatant is collected to obtain an enzymolysis solution;

[0012] Step S3, ultrafiltration, nanofiltration: The enzymatic hydrolyzate prepared in step S2 is sequentially subjected to activated carbon fiber membrane ultrafiltration and spiral membrane nanofiltration, the pH of the obtained filtrate is adjusted to 4.0-4.5 with 1 mol / L hydrochloric acid, and finally the salt NaCl is removed by ion exchange resin to obtain the soapberry peptide solution.

[0013] Step S4: Evenly mix the soapberry peptide solution with other components to obtain a soapberry peptide solution for resisting HPV infection.

[0014] Preferably, the mass ratio of soapberry meal to purified water in step S1 is 30 to 40 times.

[0015] Preferably, there is no special requirement for the source of the soapberry meal in step S1. It can be the soapberry meal which is an industrial by-product after the oil is squeezed out of the soapberry kernels, or it can be obtained by defatting the soapberry kernels.

[0016] Preferably, the method for preparing the soapberry meal in step S1 comprises the following steps:

[0017] Step D1: picking mature soapberry fruits, drying the soapberry fruits, peeling the soapberry peels to obtain hard-core seeds, cracking the outer seed shell, taking out the seed kernels, crushing them through a 40-60 mesh sieve, and setting aside;

[0018] Step D2: adding petroleum ether with a solid-liquid ratio of 1:(4-6) to defatting, extracting in a water bath shaker at 20-25° C. for 3-5 hours, separating the solid and liquid by suction filtration, repeating 2-4 times, and drying the solid phase to obtain soapberry meal.

[0019] Preferably, the mass ratio of the soapberry meal homogenate to the alkaline protease in step S2 is 100:(4-6).

[0020] Preferably, the enzymatic hydrolysis time in step S2 is 2 to 3 hours.

[0021] Preferably, the heating temperature for inactivating the enzyme in step S2 is 90-95° C. and the heating time is 10-16 min.

[0022] Preferably, the centrifugal process parameters in step S2 are: 9000-11000 r / min, room temperature, 15-20 min.

[0023] Preferably, the molecular weight cut-off of the ultrafiltration in step S3 is 3 kD.

[0024] Preferably, the mass ratio of the soapberry peptide solution and other components in step S4 is 5:(0.1-0.3).

[0025] Preferably, the other components in step S4 are made from the following raw materials in parts by weight: 8-12 parts of plant polysaccharides and 1-2 parts of zwitterionic surfactants.

[0026] Preferably, the plant polysaccharide is a mixture of mushroom polysaccharide, kelp polysaccharide and osmanthus fragrans polysaccharide in a mass ratio of (2-4):1:(1-3).

[0027] Preferably, the zwitterionic surfactant is at least one of octadecyl dimethyl betaine and dodecyl dimethyl betaine.

[0028] Another object of the present invention is to provide a soapberry peptide solution for resisting HPV infection prepared by the above-mentioned method for preparing the soapberry peptide solution for resisting HPV infection.

[0029] Another object of the present invention is to provide a use of the anti-HPV infection soapberry peptide solution in the preparation of an anti-HPV infection lotion, gel or suppository.

[0030] Due to the application of the above technical solution, the present invention has the following beneficial effects:

[0031] (1) The method for preparing the soapberry peptide solution for resisting HPV infection disclosed in the present invention has a simple process, convenient operation, high preparation efficiency and finished product qualification rate, low dependence on equipment, is suitable for continuous large-scale production, can enhance the functional added value of soapberry, has high clinical application value, and has obvious social and economic benefits.

[0032] (2) The soapberry peptide solution for resisting HPV infection disclosed in the present invention is prepared from the seeds of soapberry, a traditional Chinese medicine ingredient. The soapberry peptide can treat and prevent HPV infection, and at the same time achieve the effect of sterilization and anti-inflammation. The soapberry peptide solution prepared by this method has a low sodium content and a small peptide molecular weight, can be directly absorbed through the skin, and has a rapid effect, a definite therapeutic effect, and a low cost. From the perspective of traditional Chinese medicine, HPV infection is caused by dampness, heat, toxic evil, qi stagnation and blood stasis. Sapindus has the effect of clearing heat and dispersing blood stasis, which can effectively improve the blood circulation state of the cervix, and further effectively prevent HPV infection.

[0033] (3) The soapberry peptide solution for resisting HPV infection disclosed in the present invention can directly use the industrial by-product soapberry meal, which belongs to the recycling and reuse of solid waste, realizing the transformation of waste into treasure, which can not only reduce production costs and improve industrial economic benefits, but also be beneficial to environmental protection; the solution includes soapberry peptide solution and other components; the other components include the following raw materials in parts by weight: 8-12 parts of plant polysaccharides, 1-2 parts of zwitterionic surfactants. Through the direct mutual cooperation of the components, the combined effect can effectively improve the stability of the solution, promote skin absorption, and enhance the anti-HPV infection and antibacterial effects; the plant polysaccharide is a mixture of mushroom polysaccharide, kelp polysaccharide, and aster polysaccharide in a mass ratio of (2-4):1:(1-3). Through such a ratio and type selection, it can better play a synergistic role with the soapberry peptide solution, further improving the anti-HPV infection and antibacterial effects.

[0034] (4) The anti-HPV soapberry peptide solution disclosed in the present invention is used for the first time to prepare an anti-HPV lotion, gel, or suppository. It has significant anti-HPV efficacy and good antibacterial properties. Through the rational selection of preparation process parameters, the anti-HPV soapberry peptide solution has good transdermal properties, a significant effect, a rapid effect, and no toxic side effects. DETAILED DESCRIPTION

[0035] The following description is intended to disclose the present invention so that those skilled in the art can implement the present invention. The preferred embodiments described below are merely examples, and those skilled in the art may conceive of other obvious variations. Example 1

[0036] A method for preparing a soapberry peptide solution for resisting HPV infection comprises the following steps:

[0037] Step S1, preparation of soapberry meal homogenate: taking soapberry meal, adding it into purified water, and processing it in a high-speed tissue crushing homogenizer for 10 minutes to obtain soapberry meal homogenate;

[0038] Step S2, enzymolysis: the soapberry meal homogenate prepared in step S1 is placed in an enzymolysis tank, stirred at 88° C. for 10 minutes, and then cooled to 45° C. At this temperature, the pH is adjusted to 9 using a 0.8 mol / L NaOH solution, and alkaline protease is added thereto for enzymolysis; the NaOH solution is added during the enzymolysis process to maintain a relatively stable pH; after the enzymolysis is completed, the enzyme is inactivated by heating, the mixture is cooled, discharged, centrifuged, and the supernatant is collected to obtain an enzymolysis solution;

[0039] Step S3, ultrafiltration, nanofiltration: The enzymatic hydrolyzate prepared in step S2 is sequentially subjected to activated carbon fiber membrane ultrafiltration and spiral membrane nanofiltration, the resulting filtrate is adjusted to pH 4.0 with 1 mol / L hydrochloric acid, and finally the salt NaCl is removed by ion exchange resin to obtain the soapberry peptide solution.

[0040] Step S4: Evenly mix the soapberry peptide solution with other components to obtain a soapberry peptide solution for resisting HPV infection.

[0041] The mass ratio of soapberry meal to purified water in step S1 is 30 times.

[0042] The method for preparing the soapberry meal in step S1 comprises the following steps:

[0043] Step D1: picking mature soapberry fruits, drying the soapberry fruits, peeling the soapberry peels to obtain hard-core seeds, cracking the outer seed shell, taking out the seed kernels, crushing them through a 40-mesh sieve, and setting aside;

[0044] Step D2: adding petroleum ether with a solid-liquid ratio (mass ratio) of 1:4 to defatting, extracting in a water bath oscillator at 20°C for 3 hours, separating the solid and liquid by suction filtration, repeating twice, and drying the solid phase to obtain soapberry meal.

[0045] The mass ratio of the soapberry meal homogenate and the alkaline protease in step S2 is 100:4; the enzymatic hydrolysis time in step S2 is 2 hours; the heating temperature for inactivating the enzyme in step S2 is 90° C. and the time is 10 minutes; the centrifugal process parameters in step S2 are: 9000 r / min, room temperature, and 15 minutes.

[0046] The molecular weight cut-off of the ultrafiltration in step S3 is 3kD; the mass ratio of the soapberry peptide solution and other components in step S4 is 5:0.1; the other components in step S4 include the following raw materials in parts by weight: 8 parts of plant polysaccharides and 1 part of zwitterionic surfactant; the plant polysaccharide is a mixture of mushroom polysaccharide, kelp polysaccharide, and osmanthus fragrans polysaccharide in a mass ratio of 2:1:1; the zwitterionic surfactant is octadecyl dimethyl betaine.

[0047] A soapberry peptide solution for resisting HPV infection is prepared by adopting the preparation method of the soapberry peptide solution for resisting HPV infection.

[0048] Take the above soapberry peptide solution and use it directly as a lotion, and use it with a disposable gynecological vaginal douche to flush the cervix so that the drug can be absorbed by the cervix to achieve the purpose of treatment. Example 2

[0049] A method for preparing a soapberry peptide solution for resisting HPV infection comprises the following steps:

[0050] Step S1, preparation of soapberry meal homogenate: taking soapberry meal, adding it to purified water, and processing it in a high-speed tissue crushing homogenizer for 12 minutes to obtain soapberry meal homogenate;

[0051] Step S2, enzymolysis: the soapberry meal homogenate prepared in step S1 is placed in an enzymolysis tank, stirred at 88-92° C. for 12 minutes, and then cooled to 47° C. At this temperature, the pH is adjusted to 9.3 using a 0.9 mol / L NaOH solution, and alkaline protease is added thereto for enzymolysis; the NaOH solution is added during the enzymolysis process to maintain a relatively stable pH; after the enzymolysis is completed, the enzyme is inactivated by heating, the mixture is cooled, discharged, centrifuged, and the supernatant is collected to obtain an enzymolysis solution;

[0052] Step S3, ultrafiltration, nanofiltration: The enzymatic hydrolyzate prepared in step S2 is sequentially subjected to activated carbon fiber membrane ultrafiltration and spiral membrane nanofiltration, the pH of the obtained filtrate is adjusted to 4.2 with 1 mol / L hydrochloric acid, and finally the salt NaCl is removed by ion exchange resin to obtain the soapberry peptide solution.

[0053] Step S4: Evenly mix the soapberry peptide solution with other components to obtain a soapberry peptide solution for resisting HPV infection.

[0054] The mass ratio of soapberry meal to purified water in step S1 is 33 times.

[0055] The method for preparing the soapberry meal in step S1 comprises the following steps:

[0056] Step D1: picking ripe soapberry fruits, drying the soapberry fruits, peeling the soapberry peels to obtain hard-core seeds, cracking the outer seed shell, taking out the seed kernels, crushing them through a 45-mesh sieve, and setting aside;

[0057] Step D2: adding petroleum ether with a solid-liquid ratio of 1:4.5 to defatting, extracting in a water bath oscillator at 22°C for 3.5h, separating the solid and liquid by suction filtration, repeating 3 times, and drying the solid phase to obtain soapberry meal.

[0058] The mass ratio of the soapberry meal homogenate and the alkaline protease in step S2 is 100:4.5; the enzymatic hydrolysis time in step S2 is 2.3 hours; the heating temperature for inactivating the enzyme in step S2 is 92° C. and the time is 12 minutes; the centrifugal process parameters in step S2 are: 9500 r / min, room temperature, and 17 minutes.

[0059] The molecular weight cut-off of the ultrafiltration in step S3 is 3kD; the mass ratio of the soapberry peptide solution and other components in step S4 is 5:0.15; the other components in step S4 include the following raw materials in parts by weight: 9 parts of plant polysaccharides and 1.2 parts of zwitterionic surfactants; the plant polysaccharides are a mixture of mushroom polysaccharides, kelp polysaccharides, and osmanthus fragrans polysaccharides in a mass ratio of 2.5:1:1.5; the zwitterionic surfactant is dodecyl dimethyl betaine.

[0060] A soapberry peptide solution for resisting HPV infection is prepared by adopting the preparation method of the soapberry peptide solution for resisting HPV infection.

[0061] A soapberry peptide gel for resisting HPV infection comprises a soapberry peptide solution, carbomer, and triethanolamine. The preparation method comprises the following steps: dissolving 1.2 parts of carbomer in 50 times distilled water, allowing the solution to stand until the solution is completely swollen, adding 20 parts of the soapberry peptide solution of Example 1, stirring and mixing the mixture, adjusting the pH to 4.0 with triethanolamine, and packaging the gel. Example 3

[0062] A method for preparing a soapberry peptide solution for resisting HPV infection comprises the following steps:

[0063] Step S1, preparation of soapberry meal homogenate: taking soapberry meal, adding it to purified water, and processing it in a high-speed tissue crushing homogenizer for 13 minutes to obtain soapberry meal homogenate;

[0064] Step S2, enzymolysis: the soapberry meal homogenate prepared in step S1 is placed in an enzymolysis tank, stirred at 90° C. for 13 minutes, and then cooled to 50° C. At this temperature, the pH is adjusted to 9.5 using a 1 mol / L NaOH solution, and alkaline protease is added thereto for enzymolysis; the NaOH solution is added during the enzymolysis process to maintain a relatively stable pH; after the enzymolysis is completed, the enzyme is inactivated by heating, the mixture is cooled, discharged, centrifuged, and the supernatant is collected to obtain an enzymolysis solution;

[0065] Step S3, ultrafiltration, nanofiltration: The enzymatic hydrolyzate prepared in step S2 is sequentially subjected to activated carbon fiber membrane ultrafiltration and spiral membrane nanofiltration, the pH of the obtained filtrate is adjusted to 4.3 with 1 mol / L hydrochloric acid, and finally the salt NaCl is removed by ion exchange resin to obtain the soapberry peptide solution.

[0066] Step S4: Evenly mix the soapberry peptide solution with other components to obtain a soapberry peptide solution for resisting HPV infection.

[0067] The mass ratio of soapberry meal to purified water in step S1 is 35 times.

[0068] The method for preparing the soapberry meal in step S1 comprises the following steps:

[0069] Step D1: picking mature soapberry fruits, drying the soapberry fruits, peeling the soapberry peels to obtain hard-core seeds, cracking the outer seed shell, taking out the seed kernels, crushing them through a 50-mesh sieve, and setting aside;

[0070] Step D2: adding petroleum ether with a solid-liquid ratio of 1:5 to defatting, extracting in a water bath shaker at 23°C for 4 hours, separating the solid and liquid by suction filtration, repeating 3 times, and drying the solid phase to obtain soapberry meal.

[0071] The mass ratio of the soapberry meal homogenate and the alkaline protease in step S2 is 100:5; the enzymatic hydrolysis time in step S2 is 2.5 hours; the heating temperature for inactivating the enzyme in step S2 is 93° C. and the time is 13 minutes; the centrifugal process parameters in step S2 are: 10,000 r / min, room temperature, and 18 minutes.

[0072] The molecular weight cut-off of the ultrafiltration in step S3 is 3kD; the mass ratio of the soapberry peptide solution and other components in step S4 is 5:0.2; the other components in step S4 include the following raw materials in parts by weight: 10 parts of plant polysaccharides and 1.5 parts of zwitterionic surfactants; the plant polysaccharides are a mixture of mushroom polysaccharides, kelp polysaccharides, and osmanthus fragrans polysaccharides in a mass ratio of 3:1:2; and the zwitterionic surfactant is octadecyl dimethyl betaine.

[0073] A soapberry peptide solution for resisting HPV infection is prepared by adopting the preparation method of the soapberry peptide solution for resisting HPV infection.

[0074] A soapberry peptide suppository for preventing HPV infection is prepared as follows: 20 parts of the soapberry peptide solution of Example 1 are mixed with 4 parts of polysorbate 80, 50 parts of polyoxyethylene stearate are heated in a water bath to melt, the solution is added, stirred evenly, molded, cooled, and taken out to obtain a suppository. Example 4

[0075] A method for preparing a soapberry peptide solution for resisting HPV infection comprises the following steps:

[0076] Step S1, preparation of soapberry meal homogenate: taking soapberry meal, adding it to purified water, and processing it in a high-speed tissue crushing homogenizer for 14 minutes to obtain soapberry meal homogenate;

[0077] Step S2, enzymolysis: the soapberry meal homogenate prepared in step S1 is placed in an enzymolysis tank, stirred at 91° C. for 14 minutes, and then cooled to 53° C. At this temperature, the pH is adjusted to 9.8 using a 1.1 mol / L NaOH solution, and alkaline protease is added thereto for enzymolysis; the NaOH solution is added during the enzymolysis process to maintain a relatively stable pH; after the enzymolysis is completed, the enzyme is inactivated by heating, the mixture is cooled, centrifuged, and the supernatant is collected to obtain an enzymolysis solution;

[0078] Step S3, ultrafiltration, nanofiltration: The enzymatic hydrolyzate prepared in step S2 is sequentially subjected to activated carbon fiber membrane ultrafiltration and spiral membrane nanofiltration, the pH of the obtained filtrate is adjusted to 4.4 with 1 mol / L hydrochloric acid, and finally the salt NaCl is removed by ion exchange resin to obtain the soapberry peptide solution.

[0079] Step S4: Evenly mix the soapberry peptide solution with other components to obtain a soapberry peptide solution for resisting HPV infection.

[0080] The mass ratio of soapberry meal to purified water in step S1 is 38 times.

[0081] The method for preparing the soapberry meal in step S1 comprises the following steps:

[0082] Step D1: picking mature soapberry fruits, drying the soapberry fruits, peeling the soapberry peels to obtain hard-core seeds, cracking the outer seed shell, taking out the seed kernels, crushing them through a 55-mesh sieve, and setting aside;

[0083] Step D2: adding petroleum ether with a solid-liquid ratio of 1:5.5 to defatting, extracting in a water bath shaker at 24°C for 4.5 hours, separating the solid and liquid by suction filtration, repeating 4 times, and drying the solid phase to obtain soapberry meal.

[0084] The mass ratio of the soapberry meal homogenate and the alkaline protease in step S2 is 100:5.5; the enzymatic hydrolysis time in step S2 is 2.8 hours; the heating temperature for inactivating the enzyme in step S2 is 94° C. and the time is 15 minutes; the centrifugal process parameters in step S2 are: 10500 r / min, room temperature, 19 minutes; and the molecular weight cutoff of the ultrafiltration in step S3 is 3 kD.

[0085] The mass ratio of the soapberry peptide liquid and other components in step S4 is 5:0.25; the other components in step S4 include the following raw materials in parts by weight: 11 parts of plant polysaccharides and 1.8 parts of zwitterionic surfactants; the plant polysaccharides are a mixture of mushroom polysaccharides, kelp polysaccharides, and osmanthus fragrans polysaccharides in a mass ratio of 3.5:1:2.5; the zwitterionic surfactant is a mixture of octadecyl dimethyl betaine and dodecyl dimethyl betaine in a mass ratio of 3:5.

[0086] A soapberry peptide solution for resisting HPV infection is prepared by adopting the preparation method of the soapberry peptide solution for resisting HPV infection.

[0087] Take the above soapberry peptide solution and use it directly as a lotion, and use it with a disposable gynecological vaginal douche to flush the cervix so that the drug can be absorbed by the cervix to achieve the purpose of treatment. Example 5

[0088] A method for preparing a soapberry peptide solution for resisting HPV infection comprises the following steps:

[0089] Step S1, preparation of soapberry meal homogenate: taking soapberry meal, adding it into purified water, and processing it in a high-speed tissue crushing homogenizer for 15 minutes to obtain soapberry meal homogenate;

[0090] Step S2, enzymolysis: the soapberry meal homogenate prepared in step S1 is placed in an enzymolysis tank, stirred at 92° C. for 15 minutes, and then cooled to 55° C. At this temperature, the pH is adjusted to 10 using a 1.2 mol / L NaOH solution, and alkaline protease is added thereto for enzymolysis; the NaOH solution is added during the enzymolysis process to maintain a relatively stable pH; after the enzymolysis is completed, the enzyme is inactivated by heating, the mixture is cooled, centrifuged, and the supernatant is collected to obtain an enzymolysis solution;

[0091] Step S3, ultrafiltration, nanofiltration: The enzymatic hydrolyzate prepared in step S2 is sequentially subjected to activated carbon fiber membrane ultrafiltration and spiral membrane nanofiltration, the pH of the obtained filtrate is adjusted to 4.5 with 1 mol / L hydrochloric acid, and finally the salt NaCl is removed by ion exchange resin to obtain the soapberry peptide solution.

[0092] Step S4: Evenly mix the soapberry peptide solution with other components to obtain a soapberry peptide solution for resisting HPV infection.

[0093] The mass ratio of soapberry meal to purified water in step S1 is 40 times.

[0094] The method for preparing the soapberry meal in step S1 comprises the following steps:

[0095] Step D1: picking ripe soapberry fruits, drying the soapberry fruits, peeling the soapberry peels to obtain hard-core seeds, cracking the outer seed shell, taking out the seed kernels, crushing them through a 60-mesh sieve, and setting aside;

[0096] Step D2: adding petroleum ether with a solid-liquid ratio of 1:6 to defatting, extracting in a water bath shaker at 25°C for 5 hours, separating the solid and liquid by suction filtration, repeating 4 times, and drying the solid phase to obtain soapberry meal.

[0097] The mass ratio of the soapberry meal homogenate and the alkaline protease in step S2 is 100:6; the enzymatic hydrolysis time in step S2 is 3 hours; the heating temperature for inactivating the enzyme in step S2 is 95° C. and the time is 16 minutes; the centrifugal process parameters in step S2 are: 11000 r / min, room temperature, 20 minutes; and the molecular weight cutoff of the ultrafiltration in step S3 is 3 kD.

[0098] The mass ratio of the soapberry peptide liquid and other components in step S4 is 5:0.3; the other components in step S4 include the following raw materials in parts by weight: 12 parts of plant polysaccharides and 2 parts of zwitterionic surfactants; the plant polysaccharides are a mixture of mushroom polysaccharides, kelp polysaccharides, and osmanthus fragrans polysaccharides in a mass ratio of 4:1:3; the zwitterionic surfactant is octadecyl dimethyl betaine.

[0099] A soapberry peptide solution for resisting HPV infection is prepared by adopting the preparation method of the soapberry peptide solution for resisting HPV infection.

[0100] A soapberry peptide gel for resisting HPV infection comprises a soapberry peptide solution, carbomer, and triethanolamine. The preparation method comprises the following steps: dissolving 1.2 parts of carbomer in 50 times distilled water, allowing the solution to stand until the solution is completely swollen, adding 20 parts of the soapberry peptide solution of Example 1, stirring and mixing the mixture, adjusting the pH to 4.0 with triethanolamine, and packaging the gel.

[0101] Comparative Example 1

[0102] This example provides a soapberry peptide solution for resisting HPV infection and a preparation method thereof, which is basically the same as Example 1, except that no Osmanthus fragrans polysaccharide and zwitterionic surfactant are added.

[0103] Comparative Example 2

[0104] This example provides a soapberry peptide solution for resisting HPV infection and a preparation method thereof, which is basically the same as Example 1, except that mushroom polysaccharide is not added, and the molecular weight cutoff of the ultrafiltration in step S3 is 5kD.

[0105] In order to further illustrate the beneficial technical effects of the soapberry peptide solution for resisting HPV infection according to various embodiments of the present invention, the following experimental tests were conducted:

[0106] Experimental Example 1 Peptide Molecular Weight Distribution Detection

[0107] The peptide molecular weight distribution of the soapberry peptide solution prepared in Example 1 was tested according to the method in Appendix A of GB / T 22942-2008. The test results are as follows:

[0108]

[0109] As shown in Table 1, the peak area ratio of peptides with a molecular weight less than 500 in the soapberry peptide solution prepared in Example 1 is as high as 70.74%. Modern pharmacological studies have shown that small molecule peptides with a molecular weight less than 500 are more conducive to rapid absorption by the human body.

[0110] Experimental Example 2 Antibacterial Experiment

[0111] Test sample: the soapberry peptide solution for anti-HPV infection prepared in Example 1, take an appropriate amount of the test sample and test it in a sterile dish; test bacteria: Escherichia coli (8099) fifth generation, Staphylococcus aureus (ATCC6538) fifth generation, Candida albicans (ATCC10231) fifth generation, all of the above strains are provided by Guangdong Huankai Microbiology Technology Co., Ltd.; the inspection method and basis refer to the "Hygiene Standard for Disposable Sanitary Products" GB15979-2002 (Appendix C) C4.

[0112]

[0113] As can be seen from Table 2, the soapberry peptide solution for anti-HPV infection prepared in Example 1 has an inhibition rate of more than 99% against Escherichia coli, Staphylococcus aureus, and Candida albicans after 2 minutes of action, showing a strong antibacterial effect and meeting the relevant requirements of the "Hygiene Standard for Disposable Sanitary Products" GB15979-2002.

[0114] Experimental Example 3 Clinical Trial

[0115] 280 volunteers were recruited from the society. All of them were required to provide HPV positive test certificates from the hospital. They were divided into 7 groups on average. They were given the anti-HPV infection soapberry peptide solutions prepared in Examples 1-5 and Comparative Examples 1-2, respectively. One vial (100 mL) was used every 2 days for 1 month. The subjects' medication status was tracked regularly, and the reexamination results were collected after 1 month. The recurrence status was revisited 1 month after stopping the medication.

[0116] Precautions during medication: Take the medicine on time according to the prescribed method; use condoms when having sex during medication; stop taking the medicine if menstruation occurs during medication.

[0117] Adverse reaction assessment: If symptoms such as vulvar and vaginal itching, edema, dryness, burning sensation, redness and swelling occur, they are considered adverse reactions.

[0118] Criteria for determining efficacy

[0119] Recovered: All original subtypes have turned negative;

[0120] Markedly effective: HPV16 or HPV18 becomes negative;

[0121] Effective: HPV16 or 18 is still positive but the number of original subtypes is reduced;

[0122] Invalid: The original subtype is still positive.

[0123] The total effective rate = (cured + markedly effective + effective) / total number of cases × 100%. The test results are shown in Table 3.

[0124]

[0125] As can be seen from Table 3, the soapberry peptide solution product for anti-HPV infection according to the embodiment of the present invention has better anti-HPV infection effect than the comparative example product. The addition of osmanthus fragrans polysaccharide, zwitterionic surfactant, and mushroom polysaccharide, and the control of the ultrafiltration molecular weight cutoff at 3 kD in step S3 are all conducive to improving the therapeutic effect.

[0126] The above embodiments are only for illustrating the technical concept and features of the present invention. Their purpose is to enable people familiar with this technology to understand the contents of the present invention and implement them accordingly. They are not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A use of a soapberry peptide solution for resisting HPV infection in the preparation of a lotion, gel or suppository for resisting HPV infection, characterized in that: The anti-HPV infection soapberry peptide solution is prepared by a preparation method comprising the following steps: Step S1, preparation of soapberry meal homogenate: taking soapberry meal, adding it to purified water, and processing it in a high-speed tissue crushing homogenizer for 10 to 15 minutes to obtain soapberry meal homogenate; Step S2, enzymolysis: the soapberry meal homogenate prepared in step S1 is placed in an enzymolysis tank, stirred at 88-92° C. for 10-15 minutes, and then cooled to 45-55° C. At this temperature, the pH is adjusted to 9-10 using a 0.8-1.2 mol / L NaOH solution, and alkaline protease is added thereto for enzymolysis; the NaOH solution is added during the enzymolysis process to maintain a relatively stable pH; after the enzymolysis is completed, the enzyme is inactivated by heating, the mixture is cooled, centrifuged, and the supernatant is collected to obtain an enzymolysis solution; the mass ratio of the soapberry meal homogenate to the alkaline protease is 100:(4-6); Step S3, ultrafiltration, nanofiltration: The enzymatic hydrolyzate prepared in step S2 is sequentially subjected to ultrafiltration using an activated carbon fiber membrane and nanofiltration using a spiral membrane. The pH of the resulting filtrate is adjusted to 4.0-4.5 using 1 mol / L hydrochloric acid. Finally, the NaCl is removed through an ion exchange resin to obtain a soapberry peptide solution. The molecular weight cut-off of the ultrafiltration is 3 kD. Step S4, after uniformly mixing the soapberry peptide liquid with other components, a soapberry peptide solution for resisting HPV infection is obtained; the mass ratio of the soapberry peptide liquid to the other components is 5:(0.1-0.3); the other components include the following raw materials in parts by weight: 8-12 parts of plant polysaccharides and 1-2 parts of zwitterionic surfactants; the plant polysaccharides are a mixture of mushroom polysaccharides, kelp polysaccharides, and osmanthus fragrans polysaccharides in a mass ratio of (2-4):1:(1-3); the zwitterionic surfactant is at least one of octadecyl dimethyl betaine and dodecyl dimethyl betaine.

2. Use of the soapberry peptide solution for resisting HPV infection according to claim 1 in the preparation of lotions, gels or suppositories for resisting HPV infection, characterized in that: The mass ratio of soapberry meal to purified water in step S1 is 30 to 40 times.

3. Use of the anti-HPV infection soapberry peptide solution according to claim 1 in the preparation of an anti-HPV infection lotion, gel or suppository, characterized in that: The method for preparing the soapberry meal in step S1 comprises the following steps: Step D1: picking mature soapberry fruits, drying the soapberry fruits, peeling the soapberry peels to obtain hard-core seeds, cracking the outer seed shell, taking out the seed kernels, crushing them through a 40-60 mesh sieve, and setting aside; Step D2: adding petroleum ether with a solid-liquid ratio of 1:(4-6) to defatting, extracting in a water bath shaker at 20-25° C. for 3-5 hours, separating the solid and liquid by suction filtration, repeating 2-4 times, and drying the solid phase to obtain soapberry meal.

4. Use of the anti-HPV infection soapberry peptide solution according to claim 1 in the preparation of an anti-HPV infection lotion, gel or suppository, characterized in that: The enzymatic hydrolysis time in step S2 is 2 to 3 hours; the heating temperature for inactivating the enzyme in step S2 is 90-95° C. and the time is 10-16 minutes.

5. Use of the soapberry peptide solution for resisting HPV infection according to claim 1 in preparing lotions, gels or suppositories for resisting HPV infection, characterized in that: The centrifugal process parameters in step S2 are: 9000-11000 r / min, room temperature, 15-20 min.

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