A biorepair gel for blocking HPV virus and its preparation method

Abstract

This invention relates to the field of biomedical technology, specifically to a bio-repair gel for blocking HPV virus and its preparation method. The gel uses anhydride-modified bovine β-lactoglobulin as an antiviral active component, specifically binding to the HPV viral capsid protein to achieve physical encapsulation and blocking; recombinant type III humanized collagen as a mucosal repair component, forming a fibrous mesh scaffold to promote epithelial cell proliferation and regeneration; β-glucan as an immune-activating component, enhancing local immunity in the reproductive tract; and galactooligosaccharides as a prebiotic component, selectively nourishing lactobacilli to reshape the microecological balance. It is supplemented with a carbomer gel matrix and glycerin moisturizing agent. Through a four-component synergistic mechanism, it achieves four effects: HPV virus blocking, mucosal repair, flora regulation, and immune activation. It is suitable for HPV blocking intervention in the female reproductive tract, cervical and vaginal mucosal repair, microecological imbalance regulation, and prevention of recurrence after condyloma acuminata surgery. It falls under the category of Class II medical biological consumables.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to a bio-repair gel for blocking HPV virus and its preparation method. Background Technology

[0002] Human papillomavirus (HPV) is a type of non-enveloped, double-stranded, closed-circular small DNA virus, primarily transmitted through sexual contact, and is a major causative factor for diseases such as cervical cancer and genital warts. According to statistics from the World Health Organization, the global HPV infection rate is approximately 10%–15%. Persistent infection with high-risk HPV types (such as HPV16 / 18) is a key cause of cervical cancer, while low-risk HPV types (such as HPV6 / 11) mainly lead to benign lesions such as genital warts. Currently, clinical treatments for HPV infection and genital warts mainly include physical therapy (laser, cryotherapy, photodynamic therapy, etc.), drug therapy (interferon, imiquimod, etc.), and immunomodulatory therapy, but all have limitations to varying degrees.

[0003] Existing gel-based products for HPV protection or treatment mainly suffer from the following technical defects:

[0004] Defect 1: Limited functionality; it only blocks viruses or inhibits bacteria, lacking mucosal repair capabilities. Traditional anti-HPV gels primarily use acids or barrier agents as their main active ingredients, which can prevent HPV virus invasion for a certain period, but cannot repair mucosal damage caused by viral infection or physical therapy. Damaged mucosal barrier function is incomplete, providing conditions for secondary invasion of pathogenic microorganisms, hindering the healing of infected wounds and the clearance of HPV viruses.

[0005] Defect 2: Neglecting microecological regulation; broad-spectrum bactericides disrupt beneficial flora. Some gel products use broad-spectrum bactericides (such as triclosan and chlorhexidine) as their main active ingredients. While inhibiting pathogenic bacteria, these inevitably destroy beneficial flora such as lactobacilli that normally exist in the reproductive tract. Lactobacilli are the core flora for maintaining vaginal microecological balance. A decrease in their numbers will lead to an increase in vaginal pH and dysbiosis, which in turn creates favorable conditions for persistent HPV infection and recurrence. Clinical studies have shown that lactobacillus abundance is positively correlated with HPV clearance rate, and vaginal microecological imbalance is an important risk factor for persistent HPV infection.

[0006] Defect 3: Poor postoperative protection, resulting in a persistently high recurrence rate after genital wart treatment. The recurrence rate after physical therapy for genital warts is extremely high, reported in the literature as reaching 35%–65%. Traditional gel products lack specific design for postoperative wounds, failing to effectively promote wound healing and prevent pathogen colonization, leading to a consistently high recurrence rate. Ineffective protection and repair of the postoperative wound is one of the important reasons for the persistently high recurrence rate.

[0007] Defect 4: Poor stability of active ingredients. High-temperature sterilization processes damage protein-based active ingredients. Some new gel products are beginning to incorporate protein-based antiviral components (such as antibodies and enzymes) or active substances like collagen. However, the high-temperature sterilization step (121℃, 15-30 min) in traditional preparation processes irreversibly destroys these heat-sensitive active ingredients, significantly reducing product efficacy. How to maximize the preservation of the bioactivity of protein-based active ingredients while ensuring product sterility and safety is a pressing technical challenge that needs to be addressed.

[0008] In conclusion, there is an urgent clinical need for a bio-repair gel to block HPV viruses in order to overcome the aforementioned shortcomings of existing technologies. Summary of the Invention

[0009] To address the aforementioned technical problems, this application provides a biorepair gel for blocking HPV viruses, comprising a gel matrix, a moisturizing agent, and antiviral active components, prepared by weight from the following raw materials: 0.1-5 parts of anhydride-modified bovine β-lactoglobulin, 0.2-4 parts of recombinant type III humanized collagen, 0.5-3 parts of carbomer, 0.3-3 parts of β-glucan, 0.2-2 parts of galactooligosaccharides, 5-15 parts of glycerol, and the remainder being purified water, to a total of 100 parts.

[0010] Further improvements resulted in a final pH value of 4.0–6.0 for the gel system, which is close to the normal physiological pH range of the vagina and helps maintain the local microecological balance.

[0011] Further improvements have been made, with the finished gel product being filtered and sterilized using a 0.22μm sterile filter membrane. The bioactivity of the protein active ingredients is preserved at low temperatures throughout the entire process, without the need for high-temperature sterilization.

[0012] Further improvements were made to the mechanism of action of each component:

[0013] Anhydride-modified bovine β-lactoglobulin: The core antiviral active ingredient, after anhydride modification, changes its surface charge, exposing positively charged binding sites. It can specifically bind to the negatively charged region of the L1 / L2 region of the HPV viral capsid protein, achieving physical encapsulation of viral particles and blocking the adsorption and epithelial cell invasion of various HPV subtypes such as 6 / 11 / 16 / 18 / 31 / 33 / 45 / 52 / 58. At the same time, it can inhibit the colonization and reproduction of pathogenic bacteria such as Staphylococcus aureus, Candida albicans, and Gardnerella vaginalis, providing physical prevention and control of secondary gynecological inflammation without the risk of antibiotic resistance.

[0014] Recombinant Type III humanized collagen: a core component for mucosal repair, with nearly 100% homology to human reproductive tract mucosal tissue, excellent biocompatibility, and no rejection reaction; it can quickly fill mucosal damage caused by viral invasion and physical surgery, forming a fibrous mesh scaffold, providing three-dimensional spatial support for the crawling and proliferation of epithelial cells, accelerating wound healing, thickening the physical barrier of the reproductive tract mucosa, and reducing the probability of viral reinvasion at the body level.

[0015] β-glucan: A local immune-activating component derived from the β-1,3 / 1,6-glucan of yeast cell walls. It can target and activate the activity of immune cells such as macrophages, NK cells and dendritic cells in the reproductive tract mucosa, enhance the local innate and adaptive immunity of the reproductive tract, and help the body's own immune system to autonomously clear HPV viruses that have colonized the epithelial cells, achieving a dual antiviral effect of external blocking and internal clearing.

[0016] Galacto-oligosaccharides: A prebiotic specifically for the reproductive tract. It has a human milk oligosaccharide analog structure and cannot be absorbed and utilized by human epithelial cells. It can specifically and targetedly nourish the native beneficial lactobacilli in the reproductive tract, inhibit the excessive proliferation of harmful bacteria, quickly correct the acid-base imbalance and flora disorder of the reproductive tract, reshape the healthy micro-ecological environment of the reproductive tract, and build a strong endogenous protective barrier.

[0017] Carbomer: A medical gel matrix with excellent mucosal adhesion and sustained drug release properties. It can prolong the residence time of all active ingredients on the surface of the reproductive tract mucosa to 6-8 hours, avoid rapid loss of active ingredients, and greatly improve the utilization rate of effective ingredients. At the same time, it ensures gel formability and is suitable for non-invasive drug delivery using vaginal administration tubes.

[0018] Glycerin: A medical moisturizing and soothing agent that relieves discomfort symptoms such as dryness, stinging, and dryness of the reproductive tract mucosa caused by HPV infection and postoperative wounds, reduces the irritation of the topical gel to the damaged mucosa, and comprehensively improves the comfort of clinical use.

[0019] A method for preparing a biorepair gel for blocking HPV virus, the method comprising the following steps:

[0020] S1. Matrix pretreatment: Carbomer is swollen and dispersed in glycerol to obtain a matrix mixture;

[0021] S2. Dissolution of prebiotic components: Add β-glucan and galactooligosaccharides to purified water and stir to dissolve, thus obtaining a prebiotic solution;

[0022] S3. Addition of low-temperature active components: Add anhydride-modified bovine β-lactoglobulin and recombinant type III humanized collagen to the system under low-temperature conditions and mix evenly by low-speed stirring;

[0023] S4. pH adjustment: Adjust the pH of the system to 4.0–6.0;

[0024] S5. Sterilization filling: After filtration through a 0.22μm sterile filter membrane, the product is filled.

[0025] In a further improvement, the low-temperature condition mentioned in step S3 is that the system temperature does not exceed 25°C.

[0026] Further improvements are made to the following conditions for the swelling of carbomer in glycerol in step S1: temperature 20-25°C, time 45-60 minutes, and stirring speed 200-300 r / min; the pH adjustment in step S4 uses lactic acid or acetic acid as the regulator, with a concentration of 0.1%-0.5%.

[0027] Further improvements have been made to the application of the HPV-blocking bio-repair gel in physical interventions against HPV, repair of damaged cervical and vaginal mucosa, regulation of reproductive tract microecological imbalance, wound repair after physical therapy for condyloma acuminata, and prevention of recurrence.

[0028] The present invention has at least the following beneficial effects:

[0029] 1. This invention achieves the organic integration of four functions—virus blocking, mucosal repair, microecological regulation, and immune activation—through the scientific formulation of active components such as acid-anhydrated bovine β-lactoglobulin, recombinant type III humanized collagen, β-glucan, and galactooligosaccharides. Clinically verified, the HPV viral load reduction efficiency of this invention is 42% higher than that of a single antiviral gel, effectively filling the technological gap of existing technologies with single functions.

[0030] 2. Anhydride-modified bovine β-lactoglobulin exposes positively charged sites through anhydride modification, which can specifically bind to the negatively charged region of the HPV viral capsid protein, achieving physical encapsulation and blocking. β-glucan exerts an endogenous antiviral effect by activating local immune cells. The dual mechanisms work synergistically to achieve a 92.7% blocking rate for HPV16 / 18, and it also has good blocking effects on multiple HPV subtypes such as 6 / 11 / 16 / 18 / 31 / 33 / 45 / 52 / 58.

[0031] 3. The recombinant type III humanized collagen has nearly 100% homology with the reproductive tract mucosa tissue and can quickly form a fibrous mesh scaffold, providing three-dimensional spatial support for the crawling and proliferation of epithelial cells. Clinical data show that the mucosal healing rate of the product of this invention reaches 89.3% after 7 days, which is significantly better than the 52.1% of ordinary anti-HPV gel, effectively shortening the healing cycle of infected wounds.

[0032] 4. As a selective prebiotic, galactooligosaccharides can only be utilized by lactobacilli and not absorbed by human cells. They can effectively promote the proliferation of lactobacilli in the vagina. Clinical data show that after using the product of this invention for 14 days, the abundance of lactobacilli increased by 68.2%, the vaginal pH value returned to normal, and the imbalance of the reproductive tract microecology was effectively improved.

[0033] 5. For the wound after physical therapy for condyloma acuminata, the product of this invention integrates postoperative hemostasis, anti-infection and healing promotion. Clinical trials show that the recurrence rate of condyloma acuminata in the experimental group 3 months after surgery was 6.7%, which was significantly lower than 36.7% in the control group, and the postoperative recurrence rate was reduced by more than 38%.

[0034] 6. This invention employs a low-temperature preparation process throughout, with the addition temperature of protein-based active components controlled below 25°C. Furthermore, a 0.22μm sterile filter membrane is used to replace the traditional high-temperature sterilization process, maximizing the preservation of the bioactivity of anhydride-modified bovine β-lactoglobulin and recombinant type III humanized collagen, thus ensuring the stability of the product's efficacy.

[0035] 7. This invention does not contain hormones, antibiotics, or preservatives. It uses biocompatible natural or recombinant materials as the main active ingredients. Carbomer, as a polymer matrix, has pseudoplastic flow behavior and can remain in the vagina for 6-8 hours, achieving sustained-release administration, reducing the frequency of administration, and improving patient compliance. Attached Figure Description

[0036] Figure 1 This is a process flow diagram of the preparation method of the present invention. Detailed Implementation

[0037] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0038] Example 1: Optimal Mass Production Formulation

[0039] In this embodiment, the following raw materials were used by weight: 1.2 parts of anhydride-modified bovine β-lactoglobulin, 1.5 parts of recombinant type III humanized collagen, 1.8 parts of carbomer, 1.0 part of β-glucan, 0.8 parts of galactooligosaccharide, 10.0 parts of glycerol, and 83.7 parts of purified water.

[0040] Preparation method:

[0041] S1. Matrix pretreatment: 1.8 parts of carbomer were added to 10.0 parts of glycerol and swollen at 22℃ and 250r / min for 45 minutes to obtain a homogeneous matrix mixture.

[0042] S2. Dissolving prebiotic components: Add 1.0 part of β-glucan and 0.8 parts of galactooligosaccharide to 83.7 parts of purified water and stir at 400 r / min to dissolve, thus obtaining a prebiotic solution;

[0043] S3. Addition of active components at low temperature: Add 1.2 parts of anhydride-modified bovine β-lactoglobulin and 1.5 parts of recombinant type III humanized collagen to the system at 22°C and stir at low speed for 12 minutes to ensure that the active components are mixed evenly.

[0044] S4. pH adjustment: Adjust the pH of the system to 5.2 using a 0.3% lactic acid solution;

[0045] S5. Sterilization filling: After filtration through a 0.22μm sterile filter membrane, the product is filled.

[0046] The resulting gel product has a pH of 5.2, is uniformly translucent, has moderate viscosity, and can remain in the vagina for 6–8 hours.

[0047] Example 2: Low-concentration daily protective formula

[0048] In this embodiment, the following raw materials are taken by weight: 0.5 parts of anhydride-modified bovine β-lactoglobulin, 0.5 parts of recombinant type III humanized collagen, 1.0 part of carbomer, 0.5 parts of β-glucan, 0.4 parts of galactooligosaccharide, 8.0 parts of glycerol, and 89.1 parts of purified water.

[0049] The preparation method is the same as in Example 1, except that the pH value is adjusted to 4.5. The resulting product is suitable for daily protection of people at low risk of HPV infection and for maintaining the vaginal microecological balance.

[0050] Example 3: High-concentration post-operative repair formula

[0051] In this embodiment, the following raw materials are used by weight: 3.0 parts of anhydride-modified bovine β-lactoglobulin, 3.0 parts of recombinant type III humanized collagen, 2.5 parts of carbomer, 2.0 parts of β-glucan, 1.5 parts of galactooligosaccharide, 12.0 parts of glycerol, and 76.0 parts of purified water.

[0052] The preparation method is the same as in Example 1, except the pH value is adjusted to 5.5. The resulting product has high concentrations of anhydride-modified bovine β-lactoglobulin and collagen, making it suitable for wound repair, promoting healing, and preventing recurrence after physical therapy for condyloma acuminata.

[0053] Comparative Example 1: Galacto-oligosaccharide-free formulation

[0054] This comparative example uses the following raw materials (excluding galactooligosaccharides) by weight: 1.2 parts of anhydride-modified bovine β-lactoglobulin, 1.5 parts of recombinant type III humanized collagen, 1.8 parts of carbomer, 1.0 part of β-glucan, 10.0 parts of glycerol, and 84.5 parts of purified water.

[0055] This formula gel does not contain galactooligosaccharide prebiotic components, so it cannot play a role in regulating the flora. The abundance of lactobacilli did not increase significantly, and symptoms of flora imbalance are likely to occur after use. Compared with Example 1 which contains galactooligosaccharide, the abundance of lactobacilli did not change significantly after 14 days, and the vaginal pH value failed to return to normal, which verifies the key role of galactooligosaccharide in maintaining the balance of the microecology.

[0056] Comparative Example 2: Formula without Recombinant Collagen

[0057] This comparative example uses the following raw materials (excluding recombinant collagen) by weight: 1.2 parts of anhydride-modified bovine β-lactoglobulin, 1.8 parts of carbomer, 1.0 part of β-glucan, 0.8 parts of galactooligosaccharides, 10.0 parts of glycerol, and 85.2 parts of purified water.

[0058] This formula gel does not contain recombinant type III humanized collagen, and therefore cannot provide three-dimensional spatial support for the crawling and proliferation of epithelial cells, resulting in a significant decrease in the repair capacity of damaged mucosa.

[0059] Clinical observations showed that after 7 days of using this formula, the mucosal healing rate was only 53.2%, which was significantly different from Example 1 (89.3%) containing collagen. The probability of secondary infection was increased by about 35%, which verified the key role of recombinant collagen in mucosal repair.

[0060] Clinical efficacy test

[0061] The product of this invention has undergone the following clinical efficacy tests by an authoritative third-party testing institution:

[0062] 1. HPV blocking rate test: Using an in vitro neutralization experiment with HPV16 / 18 pseudoviruses as the test subjects, the results showed that the product of this invention had a blocking rate of 92.7% against HPV16 / 18, and also showed good blocking effects against multiple subtypes such as HPV6 / 11 / 16 / 18 / 31 / 33 / 45 / 52 / 58.

[0063] 2. Mucosal Repair Efficacy Test: 120 patients with cervical / vaginal mucosal injury were included and randomly divided into an experimental group (using the product of this invention) and a control group (using ordinary anti-HPV gel). The mucosal healing rate was observed after 7 days. The results showed that the mucosal healing rate of the experimental group was 89.3% after 7 days, which was significantly higher than that of the control group (52.1%) (P<0.01).

[0064] 3. Vaginal flora regulation effect test: Eighty patients with vaginal flora imbalance were included. After using the product of this invention for 14 days, changes in vaginal flora were analyzed by 16S rRNA gene sequencing. The results showed that the abundance of lactobacilli increased by 68.2%, the vaginal pH value returned to the normal range (3.8-4.5), and the microecological imbalance was effectively improved.

[0065] 4. Postoperative recurrence rate test of condyloma acuminata: Sixty patients who underwent physical therapy for condyloma acuminata were included and randomly divided into an experimental group (using the product of this invention, n=30) and a control group (routine care, n=30), and followed up for 3 months. The results showed that the recurrence rate in the experimental group was 6.7% (2 / 30), significantly lower than that in the control group (36.7% (11 / 30), with a postoperative recurrence rate reduction of more than 38 percentage points, verifying the significant effect of the product of this invention in preventing postoperative recurrence of condyloma acuminata.

[0066] The above clinical test data are summarized as follows:

[0067] Test Project Product of this invention control group HPV16 / 18 blocking rate 92.7% — 7-day mucosal healing rate 89.3% 52.1% Increased abundance of lactobacilli in 14 days 68.2% — Recurrence rate 3 months after surgery 6.7% 36.7%

[0068] The above clinical test data fully demonstrate that the product of this invention has excellent clinical performance in HPV virus blocking, mucosal repair, microecological regulation and postoperative recurrence prevention, and is superior to existing similar technology products.

[0069] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0070] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A bio-repair gel for blocking HPV virus, comprising a gel matrix, a moisturizing agent, and an antiviral active component, characterized in that, It is prepared from the following raw materials in parts by weight: 0.1–5 parts of anhydride-modified bovine β-lactoglobulin, 0.2–4 parts of recombinant type III humanized collagen, 0.5–3 parts of carbomer, 0.3–3 parts of β-glucan, 0.2–2 parts of galactooligosaccharides, 5–15 parts of glycerol, and the remainder is purified water, to a total of 100 parts.

2. The biorepair gel according to claim 1, characterized in that, The final pH value of the gel system is 4.0–6.

0.

3. The biorepair gel according to claim 1, characterized in that, The finished gel is sterilized by filtration through a 0.22μm sterile filter membrane, and the bioactivity of the protein active ingredients is preserved at low temperatures throughout the process without high-temperature sterilization.

4. The biorepair gel according to claim 1, characterized in that, The anhydride-modified bovine β-lactoglobulin has an anhydride modification degree of 85% to 98%. After anhydride modification, its surface charge changes, exposing positively charged binding sites. It can specifically bind to the negatively charged region of the L1 / L2 region of the HPV viral capsid protein, and block multiple subtypes of HPV virus particles such as HPV6 / 11 / 16 / 18 / 31 / 33 / 45 / 52 / 58 through physical encapsulation.

5. The biorepair gel according to claim 1, characterized in that, The recombinant type III humanized collagen has ≥99% homology with human reproductive tract mucosa tissue, and its molecular weight is 50-120kDa. It forms a fibrous network scaffold structure, providing three-dimensional spatial support for the crawling and proliferation of epithelial cells.

6. The biorepair gel according to claim 1, characterized in that, The mass ratio of β-glucan to galactooligosaccharide is 1:0.6 to 1.

2. The β-glucan is derived from β-1,3 / 1,6-glucan in yeast cell walls and is used to activate local macrophages, NK cells and dendritic cells in the mucosa. The galactooligosaccharide has a human milk oligosaccharide analog structure and selectively nourishes lactobacilli.

7. A method for preparing the biorepair gel according to any one of claims 1-6, characterized in that, Includes the following steps: S1. Matrix pretreatment: Carbomer is swollen and dispersed in glycerol to obtain a matrix mixture; S2. Dissolution of prebiotic components: Add β-glucan and galactooligosaccharides to purified water and stir to dissolve, thus obtaining a prebiotic solution; S3. Addition of low-temperature active components: Add anhydride-modified bovine β-lactoglobulin and recombinant type III humanized collagen to the system under low-temperature conditions and mix evenly by low-speed stirring; S4. pH adjustment: Adjust the pH of the system to 4.0–6.0; S5. Sterilization filling: After filtration through a 0.22μm sterile filter membrane, the product is filled.

8. The preparation method according to claim 7, characterized in that, The low-temperature condition mentioned in step S3 is that the system temperature does not exceed 25°C.

9. The preparation method according to claim 7, characterized in that, The swelling conditions of carbomer in glycerol in step S1 are: temperature 20-25℃, time 45-60 minutes, and stirring speed 200-300 r / min; the pH adjustment in step S4 uses lactic acid or acetic acid as the regulator, with a concentration of 0.1%-0.5%.

10. The use of the biorepair gel of claim 1 in the following: Physical intervention to block HPV virus, repair of damaged cervical and vaginal mucosa, regulation of reproductive tract microecological imbalance, wound repair and recurrence prevention after physical therapy for condyloma acuminata.

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