Gel dressing for preventing and treating human papilloma virus infection, preparation method and application

Abstract

The invention provides a gel dressing for preventing and treating human papilloma virus infection as well as a preparation method and application thereof, and relates to the field of pharmaceutical preparations. The gel dressing is prepared from the following raw materials: silk fibroin, triethanolamine, sodium ethyl paraben, lactoferrin, beta-glucan, a spreading hedyotis herb extract, a alternanthera philoxeroides extract, carbomer, glycerol and water. The gel dressing disclosed by the invention is excellent in effect and high in safety in the aspect of preventing and treating human papilloma virus infection.

Description

Technical Field

[0001] The invention relates to the field of pharmaceutical preparations, and in particular to a gel dressing for preventing and treating human papillomavirus infection, a preparation method and application thereof. Background Art

[0002] Human papillomavirus (HPV) can cause a variety of benign tumors or warts on human skin and mucous membranes, such as genital warts, flat warts, plantar warts and other skin and mucous membrane warts. Some types are potentially carcinogenic and cause a variety of cancers such as cervical cancer, vulvar cancer and anal cancer. The most important prevention method for HPV infection is HPV vaccination, but the vaccine is expensive and inconvenient to store. Treatments after infection include physical therapy, drug therapy, immunotherapy and surgical treatment. Among them, external application of gel dressings to prevent and treat HPV infection is a type of drug treatment. Gel dressings are suitable for reducing viral loads, preventing colonization and infection of pathogenic microorganisms, and are highly safe, effective and easy to use.

[0003] Prior art, such as patent CN113559237A, discloses a Chinese medicine gel composition for preventing and inhibiting HPV virus and its preparation method. The Chinese medicine gel composition includes, by weight, 4-5 parts of recombinant human collagen peptide, 3-6 parts of marine fish oligopeptide, 13-16 parts of lithospermum officinale, 12-14 parts of phellodendron, 12-13.5 parts of astragalus, 8-10 parts of dandelion, 9-12 parts of black plum, 8-10 parts of safflower, 7-9 parts of honeysuckle, 5-7 parts of Poria, 6-8 parts of horsetail, 3-4.5 parts of cyperus, and 0.02-0.05 parts of polyhexamethylene biguanide. The technical problem of poor antiviral effect of human papillomavirus HPV treatment drugs in the prior art is solved. The antibacterial effect is better and the side effects are smaller. However, the efficacy of the gel composition of Chinese medicine is relatively slow, and the accuracy of medication is poor.

[0004] For example, patent CN111420028A discloses a gel dressing, preparation method and application for preventing and treating human papillomavirus infection. The gel dressing is made of CP protein, carbomer, carboxylated chitosan, glycerol and gum arabic as main raw materials. The above-mentioned gel dressing uses CP protein as the main active ingredient. The CP protein is negatively charged as a whole, can bind to the HPV virus, inactivate it, and has the ability to repair damaged tissues. And patent CN117180492A discloses a gel dressing composition containing silk protein, preparation method and application. The raw materials of the composition include silk protein, carbomer, triethanolamine, sodium ethyl paraben, glycerol, cassia gelatin, Tremella polysaccharide, Eucommia male flower and water. It has the effects of nutritional repair and bioaffinity, and has obvious therapeutic effects on antibacterial and improving HPV-positive cervicitis. There is still room for improvement in the efficacy and safety of the gel dressing in the above patents.

[0005] In view of the problems existing in the prior art, it is necessary to find a gel dressing and a preparation method for preventing and treating human papillomavirus infection with better effect and stronger safety. Summary of the invention

[0006] In view of the problems existing in the prior art, the present invention provides a gel dressing for preventing and treating human papillomavirus infection, a preparation method and an application thereof. The gel dressing has excellent effect in preventing and treating human papillomavirus infection and is highly safe.

[0007] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0008] On the one hand, the present invention provides a gel dressing for preventing and treating human papillomavirus infection, the raw materials of which include silk fibroin, triethanolamine, sodium ethylparaben, lactoferrin, β-glucan, oldenlandia diffusa extract, chrysanthemum officinale extract, carbomer, glycerin and water.

[0009] Preferably, the gel dressing comprises, by weight, 0.1-2.0 parts of silk fibroin, 0.2-1.0 parts of triethanolamine, 0.05-0.5 parts of sodium ethylparaben, 0.1-2.0 parts of lactoferrin, 0.1-0.5 parts of β-glucan, 0.2-0.6 parts of oldenlandia diffusa extract, 0.2-1 parts of chrysanthemum officinale extract, 0.2-3 parts of carbomer, 5-10 parts of glycerin and 80-100 parts of water.

[0010] Further preferably, the raw materials of the gel dressing include, by weight, 0.1-1 parts of silk fibroin, 0.5-1.0 parts of triethanolamine, 0.05-0.1 parts of sodium ethylparaben, 0.1-1 parts of lactoferrin, 0.2-0.5 parts of β-glucan, 0.3-0.6 parts of oldenlandia diffusa extract, 0.3-0.6 parts of chrysanthemum officinale extract, 1-3 parts of carbomer, 6-10 parts of glycerin and 85-100 parts of water.

[0011] More preferably, the raw materials of the gel dressing include, by weight, 0.2 parts of silk fibroin, 0.8 parts of triethanolamine, 0.06 parts of sodium ethylparaben, 0.2 parts of lactoferrin, 0.4 parts of β-glucan, 0.5 parts of oldenlandia diffusa extract, 0.5 parts of chrysanthemum officinale extract, 3 parts of carbomer, 8 parts of glycerin and 90 parts of water.

[0012] Preferably, the weight ratio of the β-glucan, the Hedyotis diffusa extract and the Herba Lycopodii extract is 3-5:4-6:5-7.

[0013] Further preferably, the weight ratio of the β-glucan, the Hedyotis diffusa extract and the Herba Lycopodii extract is 4:5:5.

[0014] Preferably, the weight ratio of the silk fibroin to lactoferrin is 1:0.9-1.1; further preferably, the weight ratio of the silk fibroin to lactoferrin is 1:1.

[0015] On the other hand, the present invention also provides a method for preparing the above-mentioned gel dressing, comprising the following steps:

[0016] (1) adding glycerol to part of the water, stirring, and then adding carbomer to swell to obtain solution A;

[0017] (2) adding β-glucan, Hedyotis diffusa extract and Herba Lycopodii extract to part of the water, mixing, and then adding triethanolamine and sodium ethylparaben to obtain solution B;

[0018] (3) adding silk fibroin and lactoferrin to part of the water and dissolving them to obtain solution C;

[0019] (4) After mixing solution A and solution B, add solution C and the remaining water to obtain the product.

[0020] Preferably, the part of water in step (1) accounts for 30-40% of the total weight of water, the part of water in step (2) accounts for 20-30% of the total weight of water, and the part of water in step (3) accounts for 5-15% of the total weight of water.

[0021] Further preferably, the portion of water in step (1) accounts for 35% of the total weight of water, the portion of water in step (2) accounts for 25% of the total weight of water, and the portion of water in step (3) accounts for 10% of the total weight of water.

[0022] Preferably, the stirring speed in step (1) is 100-300 r / min, the temperature is 20-30° C., and the time is 10-30 min;

[0023] Further preferably, the stirring speed in step (1) is 200 r / min, the temperature is 25° C., and the time is 20 min;

[0024] Preferably, the stirring speed during the swelling process in step (1) is 300-500 r / min, the temperature is 70-80° C., and the time is 10-30 min.

[0025] Further preferably, the stirring speed during the swelling process in step (1) is 500 r / min, the temperature is 75° C., and the time is 30 min.

[0026] Preferably, the mixing in step (2) is ultrasonic, with a rate of 200-400 r / min, a temperature of 50-70°C, and a time of 20-30 min; further preferably, the mixing in step (2) is ultrasonic, with a rate of 300 r / min, a temperature of 60°C, and a time of 20 min.

[0027] In yet another aspect, the present invention provides use of the above-mentioned gel dressing in the preparation of a medicament for preventing and treating human papillomavirus infection.

[0028] Preferably, the human papillomavirus includes at least one of HPV1, HPV2, HPV6, HPV16, HPV18, HPV31, HPV33 and HPV35.

[0029] Further preferably, the human papillomavirus is at least one of HPV16, HPV18 and HPV35.

[0030] Compared with the prior art, the technical effects achieved by the present invention are:

[0031] 1. The gel dressing of the present invention can form a protective mucosa on the vaginal wall when in use, isolating the vaginal wall from human papillomavirus, etc., and can effectively block human papillomavirus from entering host cells through the compound between silk fibroin and lactoferrin, and then utilize the synergistic effect of β-glucan, Hedyotis diffusa extract and Herba Lysimachiae extract in the dressing raw materials, and has antiviral efficacy, and can promote the healing of damaged and eroded tissues, thereby achieving the effect of prevention and treatment. Compared with pure Chinese medicine gel dressings, the gel dressing of the present invention has stronger efficacy and more accurate medication, and compared with ordinary gel dressings, it has better safety.

[0032] 2. The gel dressing of the present invention can effectively prevent and treat human papillomavirus infection, and has a broad spectrum of efficacy and strong safety against various types of human papillomavirus (such as HPV1, HPV2, HPV6, HPV16, HPV18, HPV31, HPV33 and HPV35, etc.).

[0033] 3. The gel dressing of the present invention is easy to carry, convenient to use and painless, and the patient compliance is stronger. DETAILED DESCRIPTION

[0034] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0035] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing specific embodiments rather than for limiting the scope of protection of the present invention.

[0036] When the embodiment gives a numerical range, it should be understood that, unless otherwise specified in the present invention, the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those of ordinary skill in the art to which the present invention belongs.

[0037] It is worth noting that the raw materials used in the present invention are all common commercially available products, so their sources are not specifically limited.

[0038] Example 1

[0039] A gel dressing for preventing and treating human papillomavirus infection, wherein the raw materials include, by weight, 0.2 parts of silk fibroin, 0.8 parts of triethanolamine, 0.06 parts of sodium ethylparaben, 0.2 parts of lactoferrin, 0.4 parts of beta-glucan, 0.5 parts of oldenlandia diffusa extract, 0.5 parts of chrysanthemum officinale extract, 3 parts of carbomer, 8 parts of glycerin and 90 parts of water.

[0040] Preparation method:

[0041] (1) adding glycerol to a portion of water (accounting for 35% of the total mass of water), stirring at 200 r / min for 20 min at 25° C., then adding carbomer, stirring at 500 r / min for 30 min at 75° C. to swell, and obtaining solution A;

[0042] (2) adding β-glucan, Hedyotis diffusa extract and Herba Lycopodii extract to a portion of water (accounting for 25% of the total mass of water), ultrasonicating at 300 r / min for 20 min at 60° C., and then adding triethanolamine and sodium ethylparaben to obtain solution B;

[0043] (3) adding silk fibroin and lactoferrin to a portion of water (accounting for 10% of the total mass of water) and fully dissolving them to obtain solution C;

[0044] (4) After mixing solution A and solution B, add solution C and the remaining water to obtain a gel dressing.

[0045] Example 2

[0046] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the raw material ratio is changed, specifically as follows:

[0047] In terms of weight, the raw materials include 2.0 parts of silk fibroin, 0.2 parts of triethanolamine, 0.5 parts of sodium ethylparaben, 2.0 parts of lactoferrin, 0.1 parts of β-glucan, 0.2 parts of oldenlandia diffusa extract, 0.23 parts of chrysanthemum officinale extract, 0.2 parts of carbomer, 5 parts of glycerin and 100 parts of water.

[0048] The remaining operations are the same as those in Example 1.

[0049] Example 3

[0050] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the raw material ratio is changed, specifically as follows:

[0051] In terms of weight, the raw materials include 0.1 parts of silk fibroin, 1.0 parts of triethanolamine, 0.05 parts of sodium ethylparaben, 0.1 parts of lactoferrin, 0.5 parts of β-glucan, 0.5 parts of oldenlandia diffusa extract, 0.5 parts of chrysanthemum officinale extract, 1 part of carbomer, 10 parts of glycerin and 85 parts of water.

[0052] The remaining operations are the same as those in Example 1.

[0053] Example 4

[0054] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the parameters in the preparation method are changed, specifically as follows:

[0055] (1) adding glycerol to a portion of water (accounting for 35% of the total mass of water), stirring at 100 r / min for 10 min at 20° C., then adding carbomer, stirring at 400 r / min for 10 min at 80° C. to swell, and obtaining solution A;

[0056] (2) adding β-glucan, Hedyotis diffusa extract and Herba Lycopodii extract to a portion of water (accounting for 25% of the total mass of water), ultrasonicating at 300 r / min for 30 min at 50° C., and then adding triethanolamine and sodium ethylparaben to obtain solution B;

[0057] (3) adding silk fibroin and lactoferrin to a portion of water (accounting for 10% of the total mass of water) and fully dissolving them to obtain solution C;

[0058] (4) After mixing solution A and solution B, add solution C and the remaining water to obtain a gel dressing.

[0059] Comparative Example 1

[0060] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the raw material ratio is changed, specifically as follows:

[0061] In terms of weight, the raw materials include 3.0 parts of silk fibroin, 0.1 parts of triethanolamine, 0.01 parts of sodium ethylparaben, 3.0 parts of lactoferrin, 1.2 parts of β-glucan, 1.5 parts of oldenlandia diffusa extract, 1.5 parts of chrysanthemum officinale extract, 0.1 parts of carbomer, 12 parts of glycerin and 120 parts of water.

[0062] The remaining operations are the same as those in Example 1.

[0063] Comparative Example 2

[0064] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the raw material ratio is changed, specifically: an equal amount of lactoferrin is used to replace silk fibroin.

[0065] The remaining operations are the same as those in Example 1.

[0066] Comparative Example 3

[0067] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the raw material ratio is changed, specifically: an equal amount of silk fibroin is used to replace lactoferrin.

[0068] The remaining operations are the same as those in Example 1.

[0069] Comparative Example 4

[0070] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the raw material ratio is changed, specifically: an equal amount of CP protein is used to replace silk fibroin.

[0071] The remaining operations are the same as those in Example 1.

[0072] Comparative Example 5

[0073] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the raw material ratio is changed, specifically as follows:

[0074] In terms of weight, the raw materials include 0.2 parts of silk fibroin, 0.8 parts of triethanolamine, 0.06 parts of sodium ethylparaben, 2.5 parts of lactoferrin, 0.4 parts of β-glucan, 0.5 parts of oldenlandia diffusa extract, 0.5 parts of chrysanthemum officinale extract, 3 parts of carbomer, 8 parts of glycerin and 90 parts of water.

[0075] The remaining operations are the same as those in Example 1.

[0076] Comparative Example 6

[0077] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the raw material ratio is changed, specifically as follows:

[0078] In terms of weight, the raw materials include 0.2 parts of silk fibroin, 0.8 parts of triethanolamine, 0.06 parts of sodium ethylparaben, 0.2 parts of lactoferrin, 0.62 parts of β-glucan, 0.78 parts of chrysanthemum extract, 3 parts of carbomer, 8 parts of glycerin and 90 parts of water.

[0079] The remaining operations are the same as those in Example 1.

[0080] Comparative Example 7

[0081] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the raw material ratio is changed, specifically as follows:

[0082] In terms of weight, the raw materials include 0.2 parts of silk fibroin, 0.8 parts of triethanolamine, 0.06 parts of sodium ethylparaben, 0.2 parts of lactoferrin, 0.62 parts of β-glucan, 0.78 parts of Hedyotis diffusa extract, 3 parts of carbomer, 8 parts of glycerin and 90 parts of water.

[0083] The remaining operations are the same as those in Example 1.

[0084] Comparative Example 8

[0085] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the raw material ratio is changed, specifically as follows:

[0086] In terms of weight, the raw materials include 0.2 parts of silk fibroin, 0.8 parts of triethanolamine, 0.06 parts of sodium ethylparaben, 0.2 parts of lactoferrin, 0.7 parts of chrysanthemum officinale extract, 0.7 parts of oldenlandia diffusa extract, 3 parts of carbomer, 8 parts of glycerin and 90 parts of water.

[0087] The remaining operations are the same as those in Example 1.

[0088] Comparative Example 9

[0089] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the preparation method is changed, specifically as follows:

[0090] (1) adding glycerol to a portion of water (accounting for 35% of the total mass of water), stirring at 500 r / min for 20 min at 40° C., then adding carbomer, stirring at 800 r / min for 60 min at 45° C. to swell, and obtaining solution A;

[0091] (2) adding β-glucan, Hedyotis diffusa extract and Herba Lycopodii extract to a portion of water (accounting for 25% of the total mass of water), ultrasonicating at 100 r / min for 50 min at 30° C., and then adding triethanolamine and sodium ethylparaben to obtain solution B;

[0092] (3) adding silk fibroin and lactoferrin to a portion of water (accounting for 10% of the total mass of water) and fully dissolving them to obtain solution C;

[0093] (4) After mixing solution A and solution B, add solution C and the remaining water to obtain a gel dressing.

[0094] Comparative Example 10

[0095] A gel dressing for preventing and treating human papillomavirus infection, compared with Example 1, only the preparation method is changed, specifically as follows:

[0096] Mix all the raw materials with water, and stir at 60°C and 500 r / min for 50 minutes to obtain a gel dressing.

[0097] Test Example 1

[0098] Gel zeta potential: Gel zeta potential was measured using a Malvern potentiostat. The gel dressings prepared in Examples 1-4 and Comparative Examples 1-10 were observed at room temperature for 12 months, and tested at 0, 3, 6, 9, and 12 months. The test results are shown in Table 1.

[0099] Table 1. Gel zeta potential

[0100]

[0101]

[0102] It can be seen from the data in Table 1 that the zeta potential of the gel dressings prepared in Examples 1-4 of the present invention is negative, which can complex with the positive charge on the HPV protein capsid, thereby changing the conformation of the HPV protein and inactivating it; and the absolute value of the zeta potential of the gel dressings prepared in Examples 1-4 is significantly higher than that of the gel dressings prepared in Comparative Examples 1-10, indicating that the gel dressings obtained in the embodiments of the present invention are more stable.

[0103] Test Example 2

[0104] The therapeutic effect of the gel dressing of the present invention on cervicitis and other diseases caused by HPV infection:

[0105] 1. Purpose of clinical trial: This study aims to systematically observe the safety and effectiveness of the gel dressing of the present invention in the auxiliary treatment of chronic cervicitis caused by genital tract combined with HPV virus infection.

[0106] This clinical trial adopts a randomized, parallel control method to select subjects who meet the inclusion and exclusion criteria for the trial. The researchers select HPV-positive chronic cervicitis patients as subjects according to the inclusion and exclusion criteria. Subjects can only participate in the trial after voluntarily signing the informed consent form.

[0107] 2. Case selection:

[0108] Inclusion criteria:

[0109] (1) Aged 18-50 (inclusive), female.

[0110] (2) Meet the diagnostic criteria for chronic cervicitis.

[0111] (3) Those with positive HPV test in vaginal secretions (including those with positive HPV test in the test hospital within 10 days before screening).

[0112] (4) TCT (thin layer cytology test) negative.

[0113] (5) Voluntarily participate in the clinical trial and have signed the informed consent form.

[0114] Exclusion criteria:

[0115] (1) Patients with sexually transmitted diseases, CIN and cervical cancer.

[0116] (2) Patients with concurrent primary diseases of the heart, liver, kidney or hematopoietic system, and mental illness.

[0117] (3) Pregnant or lactating women.

[0118] (4) Those with allergic constitution or allergic to the ingredients of this product.

[0119] (5) Those who had used vaginal anti-inflammatory or antiviral drugs within one week before screening.

[0120] (6) Those who have conducted other clinical trials within 1 month.

[0121] (7) Women who have never had sexual intercourse (virgin).

[0122] (8) Patients who the researchers believe are not suitable for clinical trials.

[0123] A total of 300 cases were selected, and all had similar degrees of erosion on the cervix. There was balance among the groups in terms of age, height and weight, vital signs, degree of cervicitis (vulva, vagina and cervix), and routine examination of leucorrhea, and there was no statistical difference between the groups. The cases were divided into 15 groups, and the gel dressings prepared by Examples 1-4 and Comparative Examples 1-10 and a positive control group (PHPV fusion factor vaginal antibacterial gel, produced by Guangxi Fulaiming Biopharmaceutical Co., Ltd.) were used respectively.

[0124] 3. Dosage:

[0125] The gel dressings (vaginal packing gels) prepared in Examples 1-4 and Comparative Examples 1-10 were administered at a dosage of 3 g / time, once / day, with a course of 14 days.

[0126] The positive control group was treated with PHPV fusion factor vaginal antibacterial gel according to the instructions, 3 g / tube, once every other day, 10 times a course of treatment.

[0127] 4. Statistical analysis and evaluation methods:

[0128] (I) Statistical analysis method: SPSS20 statistical software was used to process the data, t-test was used for measurement data, and chi-square test was used for count data. P < 0.05 was considered statistically significant.

[0129] 1) Enrollment and completion status Summarize the distribution of cases in each center, including the number of enrolled cases, the number of completed cases, and the number of cases entering each analysis population, and list the list of dropped cases. Compare the total dropout rate.

[0130] 2) Baseline balance analysis describes measurement indicators such as age, height, and weight according to the measurement indicator description method. Family history, past history, allergy history, current medical history / comorbidities, etc. are described according to the counting indicator description method.

[0131] 3) Effectiveness analysis and evaluation indicators: Describe the main evaluation indicators (treatment effectiveness) of the whole center and each center, use t-test to compare the physical signs before and after treatment, and calculate whether there are significant differences before and after treatment. P<0.05 indicates statistical significance, otherwise it is considered that the treatment is ineffective.

[0132] 4) Safety analysis was described according to the counting index description method, and the inter-group comparison was analyzed by ×2 test or Fisher's exact probability method. The occurrence of complications and adverse events / reactions was described, and the specific manifestations, degree and relationship of all adverse events in each case with the device were described in detail.

[0133] 2. Clinical evaluation indicators

[0134] Effectiveness evaluation indicators:

[0135] Cured: After taking the medicine, the symptoms and signs of cervicitis such as secretions, itching, pain, congestion and edema, and erosion area basically disappeared.

[0136] Effective: After taking the medicine, the symptoms and signs of cervicitis such as secretions, itching, pain, congestion and edema, and erosion area were improved to a certain extent.

[0137] Ineffective: After taking the medicine, there was no significant improvement in symptoms and signs of cervicitis such as secretions, itching, pain, congestion and edema, and erosion area.

[0138] If the subject's efficacy is evaluated as markedly effective or effective during the treatment period, and then the treatment is terminated, the test data of the subject is valid and should be included in the statistical data, and it is not considered as dropout. SPSS20 statistical software was used to process the data, and the measurement data were tested with t test, and the counting data were tested with chi-square test. P<0.05 was considered statistically significant. One course of treatment was 14 days, and the efficacy was evaluated before and after treatment. The efficacy evaluation was performed by a specialist who was not involved in the treatment.

[0139] Safety evaluation indicators:

[0140] a. Description of safety parameters Safety parameters include: complications, adverse events / reaction incidence.

[0141] b. Methods and timing for evaluating, recording and analyzing safety parameters:

[0142] (1) Complications: According to literature reports, some patients may experience fever and fatigue, and the trial should be discontinued. Record any complications that occur during the trial.

[0143] (2) Incidence of adverse events / reactions: Record adverse events that occur during the trial and determine whether they are related to the test device or the control device, and calculate the incidence of adverse events / reactions;

[0144] The calculation formula is as follows:

[0145] Adverse event / reaction incidence rate = (number of adverse events / reactions) / (number of cases enrolled in the trial) × 100%.

[0146] 5. Clinical efficacy trial results

[0147] Case Analysis:

[0148] According to statistics, there was no significant difference in demographic data in terms of gender and ethnicity (P>0.05), age, height and weight (P>0.05), and there was no significant difference in vital signs before treatment (P>0.05). There was no difference in abnormalities in gynecological examination (vulva, vagina, cervix, uterine body, left and right adnexa) of cervicitis, see Table 2.

[0149] Table 2. Gynecological examination results for cervicitis

[0150]

[0151]

[0152] Clinical efficacy analysis:

[0153] The results of the effective evaluation of clinical efficacy and symptom improvement of cervicitis are shown in Table 3.

[0154] Table 3. Results of effective evaluation of clinical efficacy and symptom improvement in cervicitis

[0155]

[0156]

[0157] It can be seen from Table 3 that the gel dressing prepared in the embodiment of the present invention has an obvious therapeutic effect on the symptoms of cervicitis.

[0158] Analysis of HPV negative conversion rate:

[0159] Table 4. HPV negative conversion rate results

[0160] Group Positive Negative Negative conversion rate / % Example 1 2 18 90 Example 2 3 17 85 Example 3 3 17 85 Example 4 2 18 90 Comparative Example 1 14 6 30 Comparative Example 2 15 5 25 Comparative Example 3 13 7 35 Comparative Example 4 13 7 35 Comparative Example 5 14 6 30 Comparative Example 6 12 8 40 Comparative Example 7 14 6 30 Comparative Example 8 13 7 35 Comparative Example 9 12 8 40 Comparative Example 10 16 4 20 Positive control group 11 9 45

[0161] From the data in Table 4, it can be seen that the HPV negative conversion rate of the gel dressing prepared in the embodiment of the present invention is 85%-90% after one course of treatment, while that of the positive control group is 45% and that of the comparative examples 1-10 is 20%-40%, indicating that the gel dressing prepared in the embodiment of the present invention is significantly superior to the positive control group and the comparative example.

[0162] Test Example 3

[0163] Inhibitory activity detection against HPV6, HPV16 and HPV18:

[0164] 1) Construction of HPV pseudovirus

[0165] HPVL1 and L2 related genes were optimized, amplified by polymerase chain reaction (PCR), cloned into eukaryotic cell expression vectors, and then digested and sequenced to form stable expression vectors of pHPV6, pHPV16, and pHPV18. The three expression vector plasmids and reporter plasmid pSEAP (containing alkaline phosphatase reporter gene) that were correctly sequenced were amplified in large quantities and set aside.

[0166] In this experiment, DMEM high-glucose medium was used as the culture medium.

[0167] 293FT cells were passaged into 96-well plates, with 1×10 cells per well. 5 The cells were cultured at 37°C and 5% CO2 for 8 h.

[0168] 293FT cells were cultured and passaged to four 100 mm culture plates at 37°C and 5% CO2. When the cells grew to a confluency of 80%, lipoFilter transfection reagent was used to co-transfect 293FT cells with plasmids pHPV6, pHPV 16, pHPV18 and pSEAP reporter plasmid. A control group was also set up to be transfected with only pSEAP reporter plasmid. The cells transfected with the plasmid were placed in a 37°C, 5% CO2 incubator for further culture for 12 hours, and then fresh culture medium was replaced. After further culture for 72 hours, the culture medium was removed and washed twice with DPBS. The cells were digested with 0.25% trypsin for 3 minutes, 10 mL of culture medium was added and transferred into a 15 mL centrifuge tube, and the supernatant was removed after centrifugation at 1000 g for 5 minutes. An equal volume of cell lysis buffer (0.3% Brij58, 0.2% Benzonase, 0.2% plasmidsafe ATP dependent DNase in DPBS) was added to the precipitate and treated at 37°C for 24 hours. After being taken out, the cells were placed on ice for 5 minutes, 0.17 times the volume of 5M NaCl solution was added, the mixture was vortexed and placed on ice for 10-20 minutes, and the cells were centrifuged at 4°C and 5000 g for 10 minutes. The supernatant was taken, which was the pseudovirus suspension, and the tube was packaged and placed at -80°C for later use.

[0169] 2) Virus titration

[0170] 293FT cells were passaged into 96-well plates, with 1×10 cells per well. 5 The cells were cultured at 37°C and 5% CO2 for 8 h.

[0171] The HPV pseudovirus suspension was diluted 10 times continuously, from 10-1 to 10-6. The culture medium in the 96-well was removed, 50 μL of pseudovirus dilution was added, and then 50 μL of normal culture medium was added, and cultured for 72 hours at 37°C and 5% CO2. 50 μL of culture supernatant was taken and tested with Merck alkaline phosphatase test kit to calculate TCID50.

[0172] 3) Detection of the inhibitory activity of silk fibroin and lactoferrin against HPV6, HPV16 and HPV18

[0173] HPV6, HPV16, and HPV18 pseudoviruses (50 μL) of 100 TCID50 were incubated with mixed solutions of silk fibroin and lactoferrin (50 μL, mass ratio of 1:1) at different concentrations, and incubated at 37°C for 30 min; β-lactoglobulin was used as a control protein for incubation. The culture medium in the 96-well plate was removed, and the mixture after incubation was added, and the culture was continued for 12 hours at 37°C and 5% CO2. Fresh culture medium was replaced, and after 72 hours of culture, 50 μL of the culture supernatant was taken and tested using the Merck alkaline phosphatase detection kit, and the IC50 (half-maximal inhibitory concentration) of silk fibroin and lactoferrin was calculated using SPSS25. The results are shown in Table 5.

[0174] Table 5. Anti-HPV activity of silk fibroin and lactoferrin

[0175]

[0176] From Table 5, it can be seen that the specific ratio combination of silk fibroin and lactoferrin has significant inhibitory activity against HPV6, HPV16 and HPV18 viruses.

[0177] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A gel dressing for preventing and treating human papillomavirus infection, characterized in that: The raw materials include silk fibroin, triethanolamine, sodium ethylparaben, lactoferrin, β-glucan, oldenlandia diffusa extract, chrysanthemum officinale extract, carbomer, glycerin and water.

2. The gel dressing according to claim 1, characterized in that: The gel dressing comprises, by weight, 0.1-2.0 parts of silk fibroin, 0.2-1.0 parts of triethanolamine, 0.05-0.5 parts of sodium ethylparaben, 0.1-2.0 parts of lactoferrin, 0.1-0.5 parts of β-glucan, 0.2-0.6 parts of oldenlandia diffusa extract, 0.2-1 parts of chrysanthemum officinale extract, 0.2-3 parts of carbomer, 5-10 parts of glycerol and 80-100 parts of water.

3. The gel dressing according to claim 2, characterized in that: The gel dressing comprises, by weight, 0.1-1 parts of silk fibroin, 0.5-1.0 parts of triethanolamine, 0.05-0.1 parts of sodium ethylparaben, 0.1-1 parts of lactoferrin, 0.2-0.5 parts of β-glucan, 0.3-0.6 parts of oldenlandia diffusa extract, 0.3-0.6 parts of chrysanthemum officinale extract, 1-3 parts of carbomer, 6-10 parts of glycerol and 85-100 parts of water.

4. The gel dressing according to claim 3, characterized in that: The gel dressing comprises, by weight, 0.2 parts of silk fibroin, 0.8 parts of triethanolamine, 0.06 parts of sodium ethylparaben, 0.2 parts of lactoferrin, 0.4 parts of β-glucan, 0.5 parts of oldenlandia diffusa extract, 0.5 parts of chrysanthemum officinale extract, 3 parts of carbomer, 8 parts of glycerin and 90 parts of water.

5. The gel dressing according to claim 1, characterized in that: The weight ratio of the beta-glucan, the oldenlandia diffusa extract and the chrysanthemum scabra extract is 3-5:4-6:5-7.

6. The gel dressing according to claim 1, characterized in that: The weight ratio of the silk fibroin to lactoferrin is 1:0.9-1.

1.

7. The method for preparing the gel dressing according to any one of claims 1 to 6, characterized in that: The following steps are involved: (1) adding glycerol to part of the water, stirring, and then adding carbomer to swell to obtain solution A; (2) adding β-glucan, Hedyotis diffusa extract and Herba Lycopodii extract to part of the water, mixing, and then adding triethanolamine and sodium ethylparaben to obtain solution B; (3) adding silk fibroin and lactoferrin to part of the water and dissolving them to obtain solution C; (4) After mixing solution A and solution B, add solution C and the remaining water to obtain the product.

8. The preparation method according to claim 7, characterized in that: The part of water in step (1) accounts for 30-40% of the total weight of water, the part of water in step (2) accounts for 20-30% of the total weight of water, and the part of water in step (3) accounts for 5-15% of the total weight of water.

9. The preparation method according to claim 7, characterized in that: The stirring speed during the swelling process in step (1) is 300-500 r / min, the temperature is 70-80° C., and the time is 10-30 min.

10. The preparation method according to claim 7, characterized in that: The mixing in step (2) is ultrasonic, with a speed of 200-400 r / min, a temperature of 50-70° C., and a time of 20-30 min.

11. Use of the gel dressing according to any one of claims 1 to 6 in the preparation of a medicament for preventing and treating human papillomavirus infection.

12. The use according to claim 11, characterized in that: The human papillomavirus includes at least one of HPV1, HPV2, HPV6, HPV16, HPV18, HPV31, HPV33 and HPV35.

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