Use of composition gel having antibacterial and repairing functions in gynecological diseases

Abstract

Use of a composition gel having antibacterial and repairing functions in gynecological diseases, specifically a gel containing human immune cells and a cell lysis protein, which gel comprises the following components in mass concentrations: 0%-90% of human immune cells, 1%-30% of an immune cell lysis protein, 0.1%-5% of sodium hyaluronate, 0.1%-5% of natural trehalose, 0.1%-5% of glycerol, 0.1%-0.5% of salicylic acid, 0.1%-1% of a carbomer, and 0.5% of an antiviral peptide, with the balance being water. The product can remove HPV from a vagina and HPV-positive cervical cancer, improve the environment in the vagina, moisturize and maintain the vagina, promote the regeneration of the mucosal layer, and enhance the targeted immune repair for a patient, with the key being immune reconstitution. Moreover, the pH value of the final product is 6-6.5, such that the product does not affect the pH value and the environment in the vagina when being applied to the vagina, and the product is worthy of clinical popularization and application.

Description

Application of a composite gel with antibacterial and repairing functions in gynecological diseases

[0001] Cross-references

[0002] This application claims priority to the Chinese patent application filed on July 14, 2023, with application number 202310860758.1 and invention name “Application of a composition gel with antibacterial repair function in gynecological diseases”, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present application relates to the treatment of HPV-related diseases, specifically the application of a composition gel with antibacterial and repairing functions in gynecological diseases. Background Art

[0004] Cervical cancer is the second most common malignant tumor in women after breast cancer, with high morbidity and mortality rates. However, it is currently the only malignant tumor whose morbidity and mortality rates can be reduced through medical intervention. Human papillomavirus (HPV) infection rates are high among women of childbearing age, and persistent infection with high-risk HPV is the leading cause of cervical intraepithelial neoplasia (CIN) and cervical cancer. Routine cervical cancer screening, early detection of cervical HPV infection, and timely treatment are crucial for reducing the incidence and mortality of cervical cancer. Cervical cancer screening in my country started later than in the United States, and significant gaps remain in screening, diagnosis, and treatment. my country faces a severe shortage of formally trained and experienced cytology professionals, with only a handful of qualified cytologists.

[0005] Eliminating HPV and immune reconstitution are key. HPV infections remain difficult to cure primarily due to deficient HPV-targeted immune function in patients, which prevents them from effectively clearing HPV from cells, leading to long-term lesions. Immune reconstitution is crucial for clearing latent HPV from cells and strengthening targeted immune repair.

[0006] NKT cells, a new type of immune regulatory cell, are characterized by constant T cell receptor (TCR) gene expression, CD1d restriction, and rapid, high-level cytokine production. Unlike traditional T cells, NKT cells' antigen recognition is incapable of recognizing antigenic peptides presented by classic MHC class I and II molecules. Instead, they recognize lipid and protein antigens presented by CD1d molecules on the cell surface. Stimulated by these antigens, NKT cells are activated and rapidly produce cytokines such as IL4, IFNγ, IL10, and IL13, playing a crucial role in anti-tumor and anti-infection efforts, suppressing autoimmune diseases, and promoting transplantation immunity. NKT cells possess bidirectional immunomodulatory functions, both enhancing and suppressing immune responses, and are therefore closely associated with a variety of diseases, including tumors, autoimmune diseases, and transplantation tolerance. Current research has found that NKT cell numbers are reduced in patients with certain cancers (such as prostate cancer and multiple myeloma) and autoimmune diseases (such as diabetes, multiple sclerosis, rheumatoid arthritis, idiopathic thrombocytopenic purpura, and aplastic anemia). Furthermore, the ability of NKT cells to secrete IFNγ is reduced in cancer patients. The anti-tumor effects of NKT cells have been well established in mouse tumor models. Studies by David et al., which expanded NKT cells in tumor patients, and a phase I clinical trial by Ishikawa et al. in patients with advanced lung cancer demonstrated that the administration of αGalCer significantly increased the number of NKT cells, thereby exerting an anti-tumor effect. The main mechanism of NKT cell anti-tumor effects is currently believed to be: NKT cells, stimulated by their own ligands and synthetic ligands, activate the IL12R on their surface, stimulating DCs to secrete large amounts of IL12 and promoting the differentiation and maturation of immature DCs. NKT cells themselves rapidly and extensively secrete IFNγ. Simultaneously, IFNγ acts on NK cells, prompting them to secrete large amounts of IFNγ. The IL12 secreted by DCs acts on naive CD4+ T cells, polarizing them toward Th1. These factors work together to mediate the anti-tumor effect. In mouse models of autoimmune diseases (such as type 1 diabetes and experimental autoimmune meningitis), NKT cells have been shown to play a crucial role in the development and treatment of disease. By stimulating NKT cells with αGalCer, their numbers increase, and they polarize and secrete Th2 cytokines such as IL4 and IL10, thereby playing a therapeutic role in autoimmune diseases.

[0007] However, at present, there are not enough studies on the use of NKT cells to treat HPV infection, especially the research on long-acting gel-type drugs needs further development.

[0008] Summary of the Invention

[0009] On the one hand, the present application discovered that NKT cells can both enhance and suppress immune responses, thus playing an important role in anti-tumor, anti-infection, suppression of autoimmune diseases and transplant tolerance. When the HPV virus invades the human body, it activates the cellular immune mechanism, gathers at the site of infection, identifies and analyzes HPV viral DNA combinations, and synthesizes related antibodies. At the same time, the cell lysis protein is rich in a large number of immune factors, which can promote the activation and growth of systemic and local immune cells in the body, repair damaged mucosal barriers, and eliminate pathogenic microorganisms in a targeted manner. The combined immune cells achieve sensitization and efficacy, with a practical effect of 1+1 greater than 2. Active immune cells in the patient's body target and migrate deep into the deep layers of the lesion and surrounding diseased tissues to kill free and intracellular viruses in the deep infection and the surrounding area, eliminate HPV and its infected cells, correct the function of damaged cells, correct the patient's specific immune deficiency, specifically mobilize the repair function of the patient's infected skin and mucosal cells, specifically mobilize the patient's immune cell targeted killing function, repair the microenvironment around the cells, correct pathological changes, and thus block the occurrence of related diseases such as cervical cancer.

[0010] Furthermore, the present applicants have discovered that natural trehalose has physical effects such as connecting, supporting, retaining water, resisting pressure, and protecting cells. It can also affect cell survival. The addition of trehalose can enable cells or lysed proteins to survive longer in the microenvironment of the body. It can also provide nutrition to promote the implantation and proliferation of immune cells.

[0011] Furthermore, through computer simulation and screening of a large-scale screening library, the inventors obtained an antiviral peptide with excellent viral activity. The polypeptide sequence is shown in SEQ ID NO: 1 (i.e., SYKGVFWYRGSEGGK), and it has a strong inhibitory effect on HPV. The polypeptide targets the HPV E6 gene and specifically binds to E6 to form a three-dimensional structure, thereby hindering E6 from exerting its biological function. Through research, the inventors found that the polypeptide can inhibit the expression of HPV16-E6 in cervical cancer CaSki / SiHa cells, with inhibitory activity reaching 75.6% and 79.6%, respectively, demonstrating a strong inhibitory effect. Because the E6 protein is a multifunctional protein, its most important function is to mediate the degradation of the p53 protein, resulting in a weakening or even elimination of p53's inhibitory effect on cell growth and proliferation. Therefore, the function of this protein is known, and it can achieve a therapeutic effect of inhibiting HPV. Based on this, the inventors constructed a gel containing immune cells.

[0012] Furthermore, this application is implemented using the following technical solutions:

[0013] A human immune cell or cell lysate protein gel for treating vaginal HPV infection, characterized by comprising the following components at the following mass concentrations:

[0014] Human immune cells 10-90%;

[0015] Immune cell lysate protein 1%-30%;

[0016] Natural trehalose 0.1%-5%;

[0017] Sodium hyaluronate 0.1%-5%;

[0018] Glycerol 0.1% to 5%;

[0019] Salicylic acid 0.1% to 0.5%;

[0020] Carbomer 0.1% to 1%;

[0021] 0.5% of the antiviral peptide represented by SEQ ID NO: 1;

[0022] Water balance.

[0023] Furthermore, the human immune cells involved in the above human immune cells or cell lysis proteins can be human NK cells and T cells listed in the case, or can be one or more of human CIK cells, human CTL cells, human DC cells, human TIL cells, and human CAR-T cells.

[0024] Furthermore, sodium hyaluronate is a key component of the patented formula. Working together with trehalose, it improves the extracellular environment, providing essential nutritional support and environmental guarantees for cell survival. It also locks in moisture, thereby promoting new cell growth and cell metabolism, accelerating wound healing. Furthermore, sodium hyaluronate also plays an important role in the body by retaining water, maintaining the extracellular space, regulating osmotic pressure, and providing lubrication.

[0025] Furthermore, salicylic acid: an organic solvent, is used in this formula for its anti-inflammatory, antiseptic and pH regulating properties.

[0026] Furthermore, carbomer is a very commonly used additive that can reduce viscosity. It is a slightly acidic substance with a certain porosity and a strong ability to dilute substances. Therefore, when making gels, adding carbomer can reduce the viscosity of these substances and maintain the product at an appropriate viscosity. Carbomer has certain anti-inflammatory and antibacterial properties because it is a natural medicinal ingredient with certain anti-inflammatory and antibacterial effects.

[0027] Specifically, the preparation method of the vaginal gel of the present application is as follows:

[0028] First, natural trehalose, salicylic acid, carbomer, sodium hyaluronate, and glycerol are dissolved in pure water and stirred evenly. Then, purified NKT cells or human immune cell lysate protein filtered through a 0.22 μm membrane are added and stirring is continued. Then, 0.5% of the antiviral peptide shown in SEQ ID NO: 1 is added. After gentle stirring again, the remainder is supplemented with pure water. Finally, the pH value of the obtained gel is adjusted to 6.5 to obtain the finished gel. Beneficial effects

[0029] The present application relates to a commonly used pharmaceutical composition for the treatment of vaginal HPV infection and HPV-positive cervical cancer, specifically a human immune cell and cell lysis protein gel, comprising the following components in the following mass concentrations: 10%-90% human immune cells; 1%-30% immune cell lysis protein; 0.1%-5% sodium hyaluronate; 0.1%-5% natural trehalose; 0.1%-5% glycerol; 0.1%-0.5% salicylic acid; 0.1%-1% carbomer; 0.5% antiviral peptide, with the remainder being water. This product can eliminate HPV and HPV-positive cervical cancer in the vagina, improve the vaginal environment, moisturize and maintain, promote mucosal regeneration, and enhance targeted immune repair in patients, with the key being immune reconstruction. At the same time, because the final product has a pH value of 6-6.5, the product will not affect the vaginal pH and internal environment when applied to the vagina, making it worthy of clinical promotion and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] One or more embodiments are exemplarily illustrated by the accompanying figures, and these exemplary illustrations do not limit the embodiments. The word "exemplary" is used herein to mean "serving as an example, example, or illustration." Any embodiment described herein as "exemplary" is not necessarily to be construed as superior or preferred over other embodiments.

[0031] FIG1 is a micrograph of NKT cells prepared in the present application.

[0032] FIG2 is a graph showing the results of the gel anti-tumor experiment in mice of the present application. DETAILED DESCRIPTION

[0033] Those skilled in the art can refer to the contents of this article and appropriately improve the process parameters to achieve it. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art and they are all deemed to be included in this application. The methods and applications of this application have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of this application to implement and apply the technology of this application. The methods, equipment, and materials in the following implementation cases, unless otherwise specified, are conventional methods, equipment, and materials in this field and can be purchased from the market.

[0034] Example 1 Preparation of human immune cells

[0035] Specifically, the steps for isolating and culturing NKT cells are as follows:

[0036] NKT cell culture

[0037] Prepare coating solution: add 35ug Retronectin and 35ug CD16 monoclonal antibody to 7ml D-PBS and mix well.

[0038] Coating: Add the coating solution to a T75 culture flask, place it flat, mix well, and incubate at 4°C in the dark overnight.

[0039] 1.1 Blood collection: Take 50 ml of human peripheral blood and add it to a 50 ml centrifuge tube.

[0040] 1.2 Centrifugal sedimentation: 800g, 20min, room temperature. The upper layer A is the plasma layer, and the lower layer B is the Buffycoat layer and red blood cells.

[0041] 1.3 Collect the plasma layer: Gently aspirate the A layer liquid and incubate at 56°C for 30 min.

[0042] 1.4 Preparation of autologous plasma: 4°C, stand for 10 min, 1800 rpm / 800g, 4°C, 10 min, the supernatant is used as autologous plasma.

[0043] 1.5 Separation of lymphocytes with lymphocyte separation medium: Add PBS to layer B to make up to 50 ml, mix well and slowly add the volume evenly to two centrifuge tubes that have been added with 15 ml of lymphocyte separation medium. Centrifuge at 1800 rpm / 800 g, room temperature, for 20 min, no-break.

[0044] 1.6 Washing PBMCs: Use a Pasteur pipette to aspirate the white PBMC layer in the middle after centrifugation. Add about 20 ml of PBS to a new 50 ml centrifuge tube for washing. Centrifuge at 1000 rpm for 6 min. Discard the supernatant. Wash again with 20 ml of PBS. Centrifuge at 1000 rpm for 6 min. Discard the supernatant. Resuspend in 20 ml of culture medium. Mix well. Take a sample, stain and count with Typan Blue. Centrifuge at 1000 rpm for 6 min. Discard the supernatant.

[0045] 1.7 Prepare cell suspension: Resuspend the cell pellet in 40 ml of GT-T551H3 medium and mix well.

[0046] 1.8 Cleaning the culture flask: Take out the coated culture flask, discard the coating solution, wash once with 6 ml PBS, and then wash once with 6 ml culture medium.

[0047] 1.9 Inoculation of cells: Add 40 ml of cell suspension to the culture flask, add IL-15 final concentration of 30 ng / ml, IL-21 final concentration of 30 ng / ml, autologous plasma final concentration of 5%, serum substitute final concentration of 5% (US Helios product), and IL-2 final concentration of 6000 U / ml, and place the inoculated culture flask in a carbon dioxide incubator for culture.

[0048] DAY4 was cultured for 4 days and the cells were divided into 3 bottles for further culture. Each bottle was supplemented with 50 ml of GT-T551H3 medium containing IL-2500 U / ml, IL-21 at a final concentration of 30 ng / ml, IL-15 at a final concentration of 30 ng / ml, and autologous plasma at a final concentration of 2%.

[0049] On day 6, transfer the cells to a culture bag and add 500 ml of GT-T551H3 medium containing IL-2500 U / ml, IL-21 at a final concentration of 30 ng / ml, IL-15 at a final concentration of 30 ng / ml, and autologous plasma at a final concentration of 1%.

[0050] On day 8, 500 ml of GT-T551H3 medium containing IL-2500 U / ml was added, along with IL-21 at a final concentration of 30 ng / ml, IL-15 at a final concentration of 30 ng / ml, and autologous plasma at a final concentration of 1%.

[0051] On day 11, 500 ml of GT-T551H3 medium containing IL-2500 U / ml was added, along with IL-21 at a final concentration of 30 ng / ml, IL-15 at a final concentration of 30 ng / ml, and autologous plasma at a final concentration of 1%.

[0052] On day 14, 500 ml of GT-T551H3 medium containing IL-2500 U / ml was added, along with IL-21 at a final concentration of 30 ng / ml, IL-15 at a final concentration of 30 ng / ml, and autologous plasma at a final concentration of 1%.

[0053] On DAY14, 10 ml of cell suspension was collected and sent for testing for mycoplasma endotoxins and bacterial infectious diseases. No corresponding endotoxins or infectious diseases were found.

[0054] On Day 17, NKT cells were recovered. These cells function as both natural killer (NK) cells and cytotoxic T cells, expressing CD3 and CD56, as well as TCR Vα24 and TCR Vβ11. Figure 1 shows the cells during the cell culture process. As can be seen in Figure 1, NKT cells with good appearance and activity were isolated.

[0055] Example 2 Preparation of NKT immune cell lysis protein

[0056] a. Take 40 ml of the supernatant from the NKT immune cell culture prepared in Example 1 and centrifuge at 4000 × g for 20 minutes at 4°C to remove cells or cell debris (for use in step d). After centrifugation, transfer the supernatant to a new tube. Place the collected sample on ice.

[0057] b. Take 40.0 ml of the treated cell culture supernatant and place it into a 50 ml centrifuge tube. Centrifuge at 12,000 × g and 4°C for 30 min. Transfer the supernatant to another 50 ml centrifuge tube and centrifuge at 100,000 × g for 2 h. Aspirate the supernatant, taking care not to damage the cell pellet.

[0058] c. Resuspend immune cell lysate protein

[0059] Add 100ul 1×PBS and mix thoroughly by repeatedly pipetting or vortexing to completely dissolve the precipitate. This resuspension contains complete immune cell extracellular matrix protein A.

[0060] d. Preparation of NKT immune cell lysis protein combination

[0061] Freeze and thaw the NKT cells isolated in step a three times to produce cell lysate protein B. Finally, mix equal amounts of protein A and protein B and add 2 ml of saline to the volume. Store at -80°C.

[0062] Example 3 Activity Identification of Antiviral Peptides and Immune Cells and Cell Lysates

[0063] The antiviral peptide of SEQ ID NO: 1 was commissioned to be synthesized by Hongpeptide Biotechnology. The concentration of the synthesized peptide was adjusted to 1 mg / mL, and the purity was 99.97%, and it was set aside.

[0064] HPV-positive cervical cancer cell line SiHa was purchased from Shanghai Xinyu Biotechnology Co., Ltd. Cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and a 1% penicillin-streptomycin mixture at 37°C in a cell culture incubator with 5% CO2. The medium was changed every two days and cells were observed under a microscope until they reached the logarithmic growth phase. C33A cells in the logarithmic growth phase were seeded in 96-well plates. The experimental co-cultured cells were divided into 6 groups: (1) SiHa group, (2) C33A + 0.5 mg / mL antiviral peptide of SEQ ID NO: 1 group, (3) SiHa + 1 mg / mL antiviral peptide of SEQ ID NO: 1 group, (4) SiHa + NKT cell group (cell co-culture effector-target ratio of 1:10, SiHa:NKT cell) + A + B 5 mg / mL prepared in Example 2 + 1 mg / mL antiviral peptide of SEQ ID NO: 1 group, (5) SiHa + NKT cell group (cell co-culture effector-target ratio of 1:10, SiHa:NKT cell) + A + B 5 mg / mL prepared in Example 2 + 1 mg / mL antiviral peptide of SEQ ID NO: 1 group, (6) positive control group: SiHa + 5-fluorouracil 1 mg / mL. After 48 h of culture, NKT cells were removed, and 10 μL of CCK8 working solution was added to each well. The 96-well plate was placed in a 37°C incubator with 5% CO2 and incubated in the dark for 4 h. The absorbance (OD) value of each well was measured at 450 nm using a microplate reader. The results are shown in Table 1.

[0065] Table 1 CCK8 assay for cervical cancer cell proliferation in each group

[0066] The CCK8 assay was used to detect the proliferation ability of tumor cells after co-culture. The results are shown in Table 1. Compared with the SiHa cell group, the cell proliferation ability of each experimental treatment group was significantly weakened (P<0.01). As can be seen from Table 1, both the antiviral peptide of SEQ ID NO: 1 and the NKT cell combined with A+B lytic protein group can significantly reduce the proliferation activity of cancer cells and effectively inhibit HPV-positive cervical cancer cells.

[0067] Example 4 Preparation of Human Immune Cells and Cell Lysate Gel

[0068] The cells of Example 1 and the lysed protein prepared in Example 2 were prepared according to the following composition:

[0069] Human NKT immune cells 10%; cell lysis protein A+B 20%;

[0070] Sodium hyaluronate 2%; natural trehalose 1%;

[0071] Glycerol 3%;

[0072] Salicylic acid 0.5%;

[0073] Carbomer 0.5%;

[0074] 0.5% of the antiviral peptide represented by SEQ ID NO: 1;

[0075] Water balance.

[0076] Specifically, natural trehalose, salicylic acid, carbomer, sodium hyaluronate, and glycerol are first dissolved in pure water and stirred evenly. Then, the prepared NKT cells are added, and human immune cell lysis protein A+B filtered through a 0.22 μm membrane is added. After gentle stirring, a uniform system is obtained. Then, 0.5% of the antiviral peptide shown in SEQ ID NO: 1 is added, and after gentle stirring again, the remainder is supplemented with pure water. Finally, the pH value of the obtained gel is adjusted to 6.5 to obtain the finished gel.

[0077] Example 5 Preparation of Human Immune Cells and Cell Lysate Gel without Peptide Control Group

[0078] The cells of Example 1 and the lysed protein prepared in Example 2 were prepared according to the following composition:

[0079] Human NKT immune cells 10%; cell lysis protein A+B 20%;

[0080] Sodium hyaluronate 2%; natural trehalose 1%;

[0081] Glycerol 3%;

[0082] Salicylic acid 0.5%;

[0083] Carbomer 0.5%;

[0084] Water balance.

[0085] Specifically, first dissolve natural trehalose, salicylic acid, carbomer, sodium hyaluronate, and glycerol in pure water and stir evenly, then add the prepared NKT cells, add human immune cell lysis protein A+B filtered through a 0.22μm membrane, stir gently to obtain a uniform system, make up the remainder with pure water, and finally adjust the pH value of the obtained gel to 6.5 to obtain the finished gel.

[0086] Example 6 Preparation of the polypeptide control group

[0087] Prepare according to the following composition:

[0088] Sodium hyaluronate 2%; natural trehalose 1%;

[0089] Glycerol 3%;

[0090] Salicylic acid 0.5%;

[0091] Carbomer 0.5%;

[0092] 0.5% of the antiviral peptide represented by SEQ ID NO: 1;

[0093] Water balance.

[0094] Specifically, natural trehalose, salicylic acid, carbomer, sodium hyaluronate, and glycerol are first dissolved in pure water and stirred evenly. Then, 0.5% of the antiviral peptide shown in SEQ ID NO: 1 is added and stirred. The remainder is supplemented with pure water. Finally, the pH value of the obtained gel is adjusted to 6.5 to obtain the finished gel.

[0095] Example 7 Mouse Animal Experiment

[0096] The anti-tumor efficacy of the hydrogels of Examples 4-6, as well as the positive control group and the negative PBS group, was evaluated in vivo using 6-week-old female BALB / C mice. 6 SiHa cell suspension was injected subcutaneously into the axilla of the right forelimb of mice. After 2 weeks, the tumor volume reached approximately 150 mm. 3 The mice were randomly divided into 5 groups (10 mice per group). The dose of the hydrogel of Example 4-6 was 20.0 mg / kg, and it was injected only once near the tumor. The positive control group was 5-fluorouracil 20.0 mg / kg. The negative control group was an equal volume of PBS. The evaluation of the therapeutic efficacy and safety was achieved by measuring the tumor volume after 28 days and observing the survival status of the mice. The tumor volume was calculated by the following formula: V = L × W 2 / 2. L and W are the maximum and minimum diameters of the tumor, respectively.

[0097] By measuring the tumor volume, it was found that the tumor volume of the negative control group was (6352.8±56.7) mm 3 The tumor volumes of each treatment group were significantly smaller than those of the negative control group, with significant differences (P<0.01). Furthermore, as shown in Figure 2, the tumor volume of the gel group of Example 4 was the smallest (23.4±3.1) mm. 3 The effects of using cells, their lysis proteins, and antiviral peptides alone were slightly inferior to those of the gel group in Example 4, but were better than those of the positive control group. This indicates that the combination of the two has a good synergistic effect, and that they also have a good anti-cancer effect when used alone.

[0098] Observation of the groups treated with the gels of Examples 4-6 revealed no significant damage to normal organs, indicating that the local treatments lacked significant systemic toxicity. Hearts, livers, spleens, lungs, and kidneys were collected from the mice and evaluated using H&E staining. Compared to the PBS control group, the H&E staining of the gel-treated groups showed no abnormal changes, suggesting no significant damage to these organs following treatment. This demonstrates the excellent safety profile of the gels of this application.

[0099] When implementing or testing the embodiments of the present application, optional methods and materials similar to or equivalent to the methods and materials described in this specification may be used, and preferred methods, devices, and materials are described in this specification. However, before describing the materials and methods of this application, it should be understood that the specific sizes, shapes, dimensions, materials, methods, means, etc. described in this specification may be changed according to conventional experimental methods and optimization purposes, and therefore this application is not limited thereto. It should also be understood that the technical terms used in this specification are only used to describe specific types or embodiments and are not used to limit the scope of this application, which is limited only by the scope of the appended claims.

[0100] The above is only a preferred embodiment of the present application. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present application. These improvements and modifications should also be regarded as the scope of protection of the present application. Industrial Applicability

[0101] The gel containing human immune cells and cell lysis proteins provided in this application can clear HPV in the vagina and HPV in HPV-positive cervical cancer, improve the vaginal environment, moisturize and maintain, promote mucosal regeneration, and enhance the patient's targeted immune repair. The key lies in immune reconstruction; and because the pH value of the final product is 6-6.5, its application in the vagina will not affect the vaginal pH and internal environment, and is worthy of clinical promotion and application.

Claims

1. An antiviral peptide, the amino acid sequence of which is shown in SEQ ID NO: 1, which can specifically inhibit the activity of cervical cancer cells containing HPV.

2. Use of the antiviral peptide according to claim 1 in the preparation of a preparation for inhibiting the activity of cervical cancer containing HPV.

3. A gel containing human immune cells and cell lysate proteins for treating HPV, which is composed of the following components in mass concentrations: Human immune cells 10%-90%; cell lysis protein 1-30%; Sodium hyaluronate 0.1-5%; natural trehalose 0.1-5%; Glycerol 0.1% to 5%; Salicylic acid 0.1%-0.5%; Carbomer 0.1% to 1%; 0.5% of the antiviral peptide shown in SEQ ID NO: 1; Water balance; The human immune cells are NKT cells isolated from humans; the cell lysis protein is the cell lysis protein obtained by mixing the extracellular matrix obtained by NKT cell culture with NKT cell lysis solution.

4. The gel containing human immune cells and cell lysate proteins for treating HPV according to claim 3, wherein: The cell lysis protein is specifically prepared by the following method: 40 ml of NKT immune cell culture supernatant is centrifuged at 4000×g and 4°C for 20 minutes to remove cells or cell fragments, and the supernatant is aspirated into a 50 ml centrifuge tube after centrifugation, and centrifuged at 12,000×g and 4°C for 30 minutes. The supernatant is transferred to another 50 ml centrifuge tube and centrifuged at 100,000×g for 2 hours. A white precipitate is visible at the bottom of the tube, and the supernatant is aspirated without damaging the cell precipitate. 100 μl of 1×PBS is added, and the mixture is repeatedly blown or vortexed with a pipette to mix evenly to completely dissolve the precipitate. The resuspension contains complete immune cell extracellular matrix protein A; the separated NKT cells are repeatedly frozen and thawed three times to form cell lysis protein B, and finally A and B are mixed in equal amounts and added to 2 ml of saline to obtain the cell lysis protein.

5. The gel containing human immune cells and cell lysate proteins for treating HPV according to claim 3, wherein: The gel is prepared by a method comprising the following steps: First, natural trehalose, salicylic acid, carbomer, sodium hyaluronate, and glycerol are dissolved in pure water and stirred evenly. Then, the prepared NKT cells are added, and human immune cell lysate protein filtered through a 0.22 μm membrane is added. The stirring is continued to obtain a uniform system. Then, 0.5% of the antiviral peptide shown in SEQ ID NO: 1 is added and mixed evenly. Then, the remainder is supplemented with pure water, stirred evenly, and finally, the pH value of the obtained gel is adjusted to between 6 and 6.5 to obtain the finished product.

6. Use of the gel containing human immune cells and cell lysis proteins for treating HPV as described in any one of claims 3 to 5 in the preparation of a gel for inhibiting the proliferation of cervical cancer cells containing HPV.

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