A polypeptide and its use for treating HPV

Abstract

The present invention provides a polypeptide and its use for treating HPV. Further, the present invention provides the effects of specifically inhibiting the activity of HPV and promoting the apoptosis of HPV-positive cells. It was found that after using the polypeptide to treat an HPV mouse model, the growth of tumors can be significantly inhibited, and it has good application prospects.

Description

Technical Field

[0001] This application relates to the field of biology, and specifically relates to a polypeptide and its use for treating HPV. Background Art

[0002] HPV is a group of double-stranded DNA viruses belonging to the Papovaviridae family. It has a double-stranded closed-loop DNA genome, which is divided into the early region (E), the late region (L), and the long control region (LCR). The E region is divided into sub-regions 1, 2, 6, and 7. Sub-regions 6 and 7 are related to viral cell transcription, growth, and reproduction. Sub-region 2 and the LCR jointly participate in the expression of viral genes. Sub-region 1 specifically participates in viral replication and completes viral replication by affecting the activity of ATPase. Currently, about more than 110 types of HPV have been identified, and their DNA endonuclease spectra are different. More than 120 HPV genotypes have been discovered. According to the ability of the virus to promote cell carcinogenesis, they are divided into two types: low-risk HPV (LR-HPV) and high-risk HPV (HR-HPV). There are 11 types of LR-HPV: 6, 11, 40, 42, 43, 44, 54, 61, 70, 72, 51, which mainly cause benign exophytic warts and cervical intraepithelial neoplasia. There are 14 types of HR-HPV: 14, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 59, 68, 73, 82, which can mainly induce the occurrence of high-grade cervical intraepithelial neoplasia, external genital cancer, and cervical invasive squamous cell carcinoma. Types 16 and 18 are the most common among them. The positive rate of HPV16 in cervical squamous cell carcinoma is 51%, and the positive rates of HPV18 in cervical adenocarcinoma and cervical squamous cell carcinoma are 56% and 39% respectively. Many epidemiological studies have shown that the infection of HPV is very closely related to cervical lesions. Currently, it is basically considered internationally that a primary condition for the occurrence of cervical cancer and intraepithelial neoplasia is the persistent infection of high-risk HPV. According to the literature, the positive detection rates of HPV infection in LGSIL and HGSL are 64.4%-90.9% and 73.3%-100% respectively; while in cervical cancer, it is 88.4%-99.7%.

[0003] HPV has strict tissue tropism and mainly invades the squamous epithelium of the skin and mucosa. HPVDNA exists in host cells in two ways. One is the integrated state, and the other is the episomal form outside the chromosome. In pre-cancerous tissues, the virus is often located outside the chromosome, while in the cancer progression stage, the viral DNA is integrated into the host cell DNA in a single-copy or multi-copy tandem manner, resulting in the loss of the negative regulatory effect of the HPVE2 gene on the initiation of the E6 and E7 genes, and abnormal expression of the E6 and E7 genes, leading to uncontrolled cell growth.

[0004] The E6 and E7 proteins of the virus play a crucial role in the process of virus replication. These two proteins can interact with many molecular proteins, and experiments have shown that their interaction can cause unlimited proliferation and malignant transformation of cells. The E6 and E7 proteins exert their carcinogenic effects by binding to the cell cycle regulatory factors p53 and Rb proteins. The E6 protein can form a complex with the E6-associated protein (E6-AP) in the cell and specifically bind to p53. p53 is an important tumor suppressor protein that plays a negative regulatory role in the cell growth cycle. When DNA is damaged, the expression of the p53 gene is significantly upregulated. By promoting the transcription of its downstream target gene, the p21 gene, increasing the synthesis of p21, and inducing cells to arrest in the G1 phase, it exerts its role in inhibiting cell proliferation. Therefore, the specific binding of the two and their hydrolysis through the protease-dependent system cause the loss of its negative regulatory function on cell growth, resulting in the loss of normal regulation of the cell cycle and causing unlimited proliferation and malignant transformation of cells. As a multifunctional protein, it can also immortalize normal cells by activating telomerase. Studies have found that E6 also interacts with other proteins (such as target protein 1, interferon regulatory factor 3, p21, etc.) and is also related to apoptosis. The E7 protein is the main transforming protein of HPV and has a very high affinity for the retinoblastoma protein (Rb), a tumor suppressor protein related to the control of the cell cycle. The binding of E7 to Rb dissociates the Rb-E2F complex, and E2F is released to play its role as a transcription factor, resulting in the loss of control of the cell cycle and immortalization. After HPV infects the body, it can simultaneously cause humoral and cellular immune responses. IgG antibodies can be detected in the bodies of some infected patients. These antibodies mainly target the viral capsid proteins and have virus type and structure specificities. Such neutralizing antibodies can only bind to free viruses outside the cell, inhibit the adhesion of the virus to epithelial cells and its penetration into the cells, so they can only prevent new infections. However, for infected host cells, especially epithelial basal cells, the expression of the antigen encoded by the L1 gene will be significantly reduced after infection, and the corresponding antibody titer is low, which is not sufficient to cause antibody-mediated cell killing. Therefore, humoral immunity can only be regarded as a response to HPV infection but has no ability to eliminate HPV infection.

[0005] There is currently a lack of clinically recognized effective treatments for cervical HPV infection. In recent years, the clinical treatment of cervical HPV infection mainly includes drugs, physical therapy, surgery and comprehensive treatment. Broad-spectrum antiviral preparations have multiple effects such as inhibiting RNA virus replication, regulating immunity, antiviral and anti-tumor, and are currently the most important antiviral and anti-tumor biological products. Studies have shown that interferon kappa induces sp100 protein to inhibit the transcription of human papillomavirus 31. The results show that interferon has anti-human papillomavirus activity and can quickly inhibit HPV transcription. There are also studies that use RevMan5.3 software to conduct statistical analysis on 3442 patients in 23 literatures, and found that interferon has significant effects on the treatment of HPV infection and good safety. Virus-like protein particle vaccines (VLPs) are the earliest developed and most mature vaccines, and there is no obvious cross-protection effect between different subtype vaccines. In order to expand the coverage of various HPV subtypes, VLPs vaccines are usually designed as multivalent vaccines that can prevent multiple, especially high-risk HPV infections at the same time. Currently, there are three main vaccines currently on the market, including the 2-valent vaccine Cervarix (preventing HPV16 and 18 subtypes) produced by GSK, the 4-valent vaccine Gardasil (preventing HPV6, 11, 16, 18 subtypes) produced by Merck, and the 9-valent vaccine V503 (preventing HPV6, 11, 16, 18, 31, 33, 45, 52, 59 subtypes). The relatively mature VLPs vaccines independently developed in China are also being developed in an orderly manner. There are many methods in the literature on the treatment of cervical HPV infection with traditional Chinese medicine. Oral Chinese medicine decoctions are used to strengthen the body's immune system; local application of medicines is used to clear away heat and toxicity, remove rot and promote new growth, and eliminate HPV infection. Researchers found that 160 patients with high-risk cervical HPV infection were treated with Chinese medicine combined with Baofukang suppositories. The results showed that the clinical efficacy of Chinese medicine decoctions combined with external use of Baofukang for patients with high-risk cervical HPV infection was acceptable.

[0006] Based on the above analysis, it is found that there are many drugs for the clinical treatment of cervical HPV virus infection, but there is currently a lack of recognized effective specific drugs. Therefore, the development of more effective drugs for the treatment of HPV virus is an important research direction. Summary of the invention

[0007] Studies have found that caerin polypeptide has a killing effect on HPV+ cells in vivo and in vitro, but the killing effect is not strong enough and needs further optimization and improvement.

[0008] The present invention is a Caerin1.9 polypeptide (GLFGVLGSIAKHVLPHVVPVIAEKL-NH 2On the basis of [[ID=]], through bioinformatics analysis, it was found that the 5th amino acid V was modified to C, the 20th V was modified to H, and myr modification was carried out at the N-terminus. Leon Biology was commissioned to synthesize the modified sequence, and the synthesized sequence was myr-GLFGCLGSIAKHVLPHVVPHIAEKL-NH 2 , named Caerin-mGJ. By such a modification method, the ability of the polypeptide to penetrate cell membranes and its killing activity can be improved, and the activity is relatively high.

[0009] Specifically, the present invention provides a pharmaceutical composition for treating HPV infection, and the pharmaceutical composition contains the Caerin-mGJ polypeptide, and its amino acid sequence is as

[0010] myr-GLFGCLGSIAKHVLPHVVPHIAEKL-NH 2 shown.

[0011] More specifically, the present invention also provides the use of the Caerin-mGJ polypeptide in the preparation of a drug for treating HPV infection, wherein the Caerin-mGJ polypeptide, its amino acid sequence is as

[0012] myr-GLFGCLGSIAKHVLPHVVPHIAEKL-NH 2 shown.

[0013] Furthermore, the present invention also provides a method for inhibiting HPV in vitro, and the method includes the step of contacting the Caerin-mGJ polypeptide with target cells, wherein the Caerin-mGJ polypeptide, its amino acid sequence is myr-GLFGCLGSIAKHVLPHVVPHIAEKL-NH 2 shown.

[0014] Furthermore, the drug or pharmaceutical composition of the present invention also includes a pharmaceutically acceptable carrier.

[0015] When a therapeutically effective amount of the polypeptide of the present invention is designed to be administered, for example, by intravenous, cutaneous, or subcutaneous injection, the binder will be in the form of a pyrogen-free, parenterally acceptable aqueous solution. Considering appropriate pH, isotonicity, stability, etc., methods for preparing parenterally acceptable protein solutions are within the skill in the art. In addition to the binder, preferred pharmaceutical compositions for intravenous, cutaneous, or subcutaneous injection will contain an isotonic vehicle such as sodium chloride injection, Ringer's injection, glucose injection, glucose and sodium chloride injection, Ringer's injection containing lactate, or other vehicles known in the art. The pharmaceutical composition of the present invention may also contain stabilizers, preservatives, buffers, antioxidants, or other additives well known to those skilled in the art.

[0016] Pharmaceutically acceptable excipients used in the preparation of the provided pharmaceutical compositions include inert diluents, dispersing and / or granulating agents, surfactants and / or emulsifying agents, disintegrants, binders, preservatives, buffering agents, lubricants and / or oils. Excipients (such as cocoa butter and suppository waxes), colorants, coating agents, sweetening agents, flavoring agents, and aromatic agents may also be present in the composition.

[0017] Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dibasic calcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dried starch, corn starch, powdered sugar, and mixtures thereof.

[0018] Exemplary granulating and / or dispersing agents include potato starch, corn starch, tapioca starch, sodium starch glycolate, clay, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponges, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, croscarmellose sodium (cross-linked carboxymethyl cellulose), methyl cellulose, pregelatinized starch (starch 1500), microcrystalline starch, water-insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof.

[0019] Exemplary surfactants and / or emulsifiers include natural emulsifiers (e.g., gum arabic, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan gum, pectin, gelatin, egg yolk, casein, lanolin, cholesterol, waxes, and lecithin), bentonites (e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminum silicate)), long-chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glycerol monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic polymers, and carboxyvinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethyl cellulose, powdered cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan, polyoxyethylene sorbitan monooleate, sorbitan monopalmitate, sorbitan monostearate, sorbitan tristearate, glycerol monooleate, sorbitan monooleate, polyoxyethylene esters (e.g., polyoxyethylene monostearate, polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, sucrose fatty acid esters, polyethylene glycol fatty acid esters, polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether, poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, poloxamer P-188, cetrimide, cetylpyridinium chloride, benzalkonium chloride, sodium docusate, and / or mixtures thereof.

[0020] Exemplary binders include starches (e.g., corn starch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., gum arabic, sodium alginate, Irish moss extract, panwar gum, ghatti gum, mucilage of isabgol husk, carboxymethyl cellulose, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate, and larch arabinogalactan, alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, ethanol, and / or mixtures thereof.

[0021] Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, antiprotozoal preservatives, alcohol preservatives, acidic preservatives, and other preservatives. In some embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent.

[0022] Exemplary antioxidants include α-tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, thioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.

[0023] Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and its salts and hydrates (e.g., sodium ethylenediaminetetraacetate, disodium ethylenediaminetetraacetate, trisodium ethylenediaminetetraacetate, calcium disodium ethylenediaminetetraacetate, dipotassium ethylenediaminetetraacetate, etc.), citric acid and its salts and hydrates (e.g., citric acid monohydrate), fumaric acid and its salts and hydrates, malic acid and its salts and hydrates, phosphoric acid and its salts and hydrates, and tartaric acid and its salts and hydrates. Exemplary antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethanol, glycerol, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.

[0024] Exemplary antifungal preservatives include butyl paraben, methyl paraben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid.

[0025] Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, parabens, and phenylethyl alcohol.

[0026] Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E, β-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.

[0027] Regarding the administration of the pharmaceutical composition of the present invention, the dosage range will generally be about 0.0001 - 100 milligrams per kilogram (mg / kg) of the host body weight, and more typically 0.01 - 5 mg / kg of the host body weight. Exemplary dosages can be 0.25 mg / kg body weight, 1 mg / kg body weight, 3 mg / kg body weight, 5 mg / kg body weight, or 10 mg / kg body weight, or within the range of 1 - 10 mg / kg. An exemplary treatment regimen is administration once or twice a day, or once or twice a week, once every two weeks, once every three weeks, once every four weeks, once a month, once every two or three months, or once every three to six months. The dosage can be selected and readjusted by a skilled healthcare professional as needed to maximize the therapeutic benefit for a particular patient.

[0028] The pharmaceutical composition of the present invention will typically be administered to the same patient in a variety of situations. The interval between single doses can be, for example, 2 - 5 days, weekly, monthly, every two or three months, every six months, or annually. Based on adjusting the blood level or other markers in the subject or patient, the interval between administrations can also be irregular. The dosage regimen of the compound of the present invention includes intravenous administration at 1 mg / kg body weight or 3 mg / kg body weight, wherein the compound is administered once every two to four weeks for up to six doses, and then once every three months at 3 mg / kg body weight or 1 mg / kg body weight.

[0029] Beneficial effects

[0030] The present invention provides a polypeptide and its use for treating HPV. Further, the present invention provides the effects of specifically inhibiting HPV activity and promoting apoptosis of HPV-positive cells. After using the polypeptide in the treatment of an HPV mouse model, it was found that it can significantly inhibit tumor growth, and has good application prospects. Description of the drawings

[0031] Figure 1 Effects of each treatment group on the apoptosis rate Detailed implementation manners

[0032] The specific embodiments of the present invention will be described in more detail below with reference to the drawings. Although the specific embodiments of the present invention are shown in the drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to enable a more thorough understanding of the present invention and to fully convey the scope of the present invention to those skilled in the art.

[0033] Example 1 Improvement and modification of Caerin 1.9 polypeptide

[0034] According to the sequence of Caerin1.9 polypeptide (GLFGVLGSIAKHVLPHVVPVIAEKL-NH 2 ), through bioinformatics analysis, it was found that the 5th amino acid V was modified to C, the 20th V was modified to H, and at the same time, myr modification was carried out at the N-terminus. Leon Biosciences was commissioned to synthesize the modified sequence, and the synthesized sequence was myr-GLFGCLGSIAKHVLPHVVPHIAEKL-NH 2 , named Caerin-mGJ. At the same time, GLFGCLGSIAKHVLPHVVPHIAEKL-NH 2 was also synthesized and named Caerin-GJ. Through such a modification method, the ability of the polypeptide to penetrate the cell membrane and its killing activity can be improved, and the activity is relatively high. Caerin1.9 polypeptide, myr-Caerin1.9 polypeptide and the control polypeptide Caerin-DZ: myr-GLFGSLGSIAKHVLPHVVPRIAEKL-NH 2 were also respectively commissioned to Leon Biosciences for biosynthesis. HPLC was used to identify that their purity was above 95%, and the polypeptide concentration was adjusted to 2 mg / mL for standby.

[0035] Example 2 Activity Identification of Each Modified Polypeptide of Caerin

[0036] HPV+TC-1 cell line, product number Cell-1201, was purchased from Shanghai Fengshou Biotechnology Co., Ltd. (HPV+TC-1 cells are C57BL / C(H-2b) mouse lung epithelial cells co-transformed with HPV16 E6, E7 and ras genes). HUVEC cells, product number hz10729, were purchased from Shanghai Hzhen Biotechnology Co., Ltd.

[0037] TC-1 and HUVEC cells in the logarithmic growth phase were selected for MTT assay. The cells in the culture flask were digested and centrifuged, and the cell suspension concentration was adjusted to 5×10 5 cells / ml. 100 μl of cell suspension was evenly inoculated into each well of a 96-well plate (i.e., 5×10 3 cells per well), and the outer peripheral wells were filled with sterile PBS. It was placed in a 5% CO 2 , 37 °C incubator and incubated for 24 h, and the cell monolayer could cover the bottom of the well (96-well flat bottom plate). The polypeptide drug groups were set up, and a blank control group (containing only PBS) with 3 replicates in each group was set up, and each experiment was repeated three times. The polypeptide drug groups were Caerin1.9 polypeptide, myr-Caerin1.9 polypeptide, control polypeptide Caerin-DZ, and Caerin-GJ polypeptide with a concentration of 15 μg / ml respectively. Each group was placed in 5% CO 2, incubate again at 37°C for 24 hours. Add 10μl of MTT solution (5mg / ml, i.e. 0.5% MTT) to each well and place in the incubator for another 4 hours. Terminate the culture and carefully remove the liquid in the wells. Add 150μl of dimethyl sulfoxide (DMSO) to each well and place on a shaker for 10 minutes at a low speed to fully dissolve the crystals. Use an enzyme-linked immunosorbent assay to detect the absorbance of each well at an OD value of 570nm. Cell survival rate = (OD value of the drug well - OD value of the blank well) / (OD value of the untreated well - OD value of the blank well) × 100%. The results are shown in Table 1.

[0038] Table 1 Effects of each group on the cell viability of HPV+TC-1 cells and HUVEC cells

[0039] Group Survival rate of HPV+TC-1 cells (%) Survival rate of HUVEC cells (%) Blank control group 100.00±1.30 100.00±1.91 Caerin1.9 polypeptide group 19.29±0.31* 94.17±2.56 myr-Caerin1.9 group 15.37±0.26* 93.88±2.17 Caerin-DZ control group 70.57±0.85 95.89±3.24 Caerin-mGJ polypeptide group 5.53±0.08* 97.63±2.87 Caerin-GJ polypeptide group 9.88±0.12* 97.88±2.13

[0040] From the results in Table 1, it can be seen that Caerin1.9 polypeptide, myr-Caerin1.9 polypeptide and Caerin-GJ polypeptide have an inhibitory effect on the proliferation of TC-1 cells, and the difference is very significant (*P<0.01). From the results, it can be seen that myr modification can improve the cell permeability of the polypeptide and improve the biological activity of the polypeptide. The modified mutant polypeptide Caerin-mGJ polypeptide of the present invention has a very significant activity improvement effect. At the same time, it has no cytotoxic effect on normal cells.

[0041] Example 3 Apoptotic Activity of Caerin-mGJ Polypeptide

[0042] The method is a conventional method in the art. Specifically, the HPV+TC-1 cells with vigorous proliferation in Example 2 were selected for apoptosis experiment. First, the TC-1 cells were digested and centrifuged to adjust the cell concentration to 10 6 cells / ml, 1 ml of cell suspension was inoculated into each well of a 24-well flat-bottom plate. Each experiment was repeated three times. Overnight culture: placed in 5% CO 2 , incubated in a 37°C incubator for 24 hours, and observed under a microscope that the cells covered the bottom of the well. Use peptide treatment: set one well as no treatment, and the remaining wells as the experimental group, add different concentrations (15, 10 and 5μg / ml) of Caerin-mGJ peptide solution, and use Caerin-GJ peptide and Caerin1.9 peptide as controls, with the concentrations of 10μg / ml. Place again in 5% CO 2 , incubated at 37°C for 24 hours. Collect the cell supernatant, digest and centrifuge the TC-1 cells in each well, wash them twice with phosphate buffered saline (PBS) containing 2% fetal bovine serum (FBS), count the number of cells in each group using a cell counter, and then divide each group into three samples and include them in the experimental specimens. Adjust the number of cells in the flow tube to 106 Cells were centrifuged at 1000 rpm for 5 min, the supernatant was removed, and the cells were resuspended in 100 μl of Binding Buffer. All samples were stained: in the reference samples, 1 tube of Unstained cells (without any antibody labeling), 1 tube was added with 5 μl of AnnexinV-FITC labeling, 1 tube was added with 5 μl of PI labeling, and the remaining samples were simultaneously added with 5 μl of AnnexinV-FITC and 5 μl of PI, incubated in the dark at room temperature for 15 min, and then 400 μl of Binding Buffer was added to terminate the reaction. They were washed twice with PBS containing 2% FBS, and finally resuspended in 200 μl of PBS containing 2% FBS. The apoptosis rate of the cells was detected by flow cytometry, and the obtained results were analyzed and processed by Flowjo V10 software. The results are as Figure 1 shown.

[0043] From Figure 1 the results, it can be seen that after the HPV+TC-1 cell line was treated with the polypeptide for 24 h, compared with the blank control group, Caerin-mGJ polypeptide could significantly promote the apoptosis of TC-1 cells and inhibit the proliferation of TC-1 cells (P<0.01). With the increase of the concentration, the percentage of Caerin-mGJ polypeptide promoting tumor cell apoptosis increased, indicating a certain dose-dependence. When the concentration was 15 μg / ml, the apoptosis rate reached (95.13±2.83)%, and the effect was better.

[0044] Example 3 Animal model experiment of the polypeptide

[0045] The TC-1 tumor mouse model was used to explore the in vivo anti-tumor activity of the polypeptide, and the method also referred to the mature methods in the art. Specifically, on the 4th to 5th day after subcutaneous inoculation of TC-1 cells into C57 mice, mice with a tumor size diameter of approximately 5 mm were selected for the experiment. The specific tumor-bearing mice were completely randomized into groups using a random number table: control group: PBS group, experimental group 1: Caerin-mGJ polypeptide group, experimental group 2: Caerin-GJ polypeptide group; positive control group: Caerin1.9 polypeptide group. The administration method was intratumoral injection. The PBS group was given 100 μl of PBS, and each experimental group was given 100 μl of a polypeptide solution containing 30 μg. Injection was carried out continuously for 7 days. On the 14th day after the injection was completed, the long diameter and short diameter of the tumor were measured. The tumor volume was calculated (tumor volume = 1 / 2×long diameter×short diameter2). The results are shown in Table 2.

[0046] Table 2 Results of the effects of each group on tumor volume

[0047] Group <![CDATA[Tumor volume (mm 3 )]]> Control group 689.52±48.61 Experimental group 1 79.56±5.57## Experimental group 2 115.89±10.28# Positive control group 205.41±15.67#

[0048] As can be seen from Table 2, compared with the tumor size of the mice in the control group, there were significant statistical differences in the tumor size of the tumor-bearing mice treated with the polypeptides in each experimental group (P<0.01). In particular, the tumor suppression effect of Experimental Group 1 was the most significant, with a significant decrease in volume and extremely strong application prospects.

[0049] From the tumor-bearing model of mice, it can be seen that the cervical cancer model can be effectively treated by the polypeptides, and those skilled in the art can confirm that the polypeptides can be used for the treatment of cervical cancer caused by HPV.

[0050] Finally, it should be noted that: the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A polypeptide capable of treating HPV, characterized in that The polypeptide is a Caerin-mGJ polypeptide, and its amino acid sequence is shown as myr-GLFGCLGSIAKHVLPHVVPHIAEKL-NH2.

2. Use of the Caerin-mGJ polypeptide as claimed in claim 1 in the preparation of a drug for cervical cancer infected by HPV.

3. Use of the Caerin-mGJ polypeptide as claimed in claim 1 in the preparation of an agent for inhibiting HPV proliferation and promoting apoptosis of cervical cancer in vitro.

4. A method for inhibiting HPV in vitro, the method comprising the step of contacting a Caerin-mGJ polypeptide with a target cell containing HPV, wherein: The amino acid sequence of Caerin-mGJ polypeptide is shown as myr-GLFGCLGSIAKHVLPHVVPHIAEKL-NH2.

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