Application of ailanthus altissima milk gel in preparation of drugs for resisting low-risk HPV infection and preventing and treating low-risk HPV related diseases

An HPV virus and disease technology, applied in drug combinations, skin diseases, antiviral agents, etc., can solve the problems of reducing recurrence rate, lack of antiviral drugs, and lack of drugs for early control of condyloma acuminatum

Pending Publication Date: 2022-05-10
ZHUZHOU QIANJIN PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although most studies believe that traditional physical methods to remove lesions and combined with antiviral therapy can effectively control the development and recurrence of condyloma acuminata, but there is still a lack of effective antiviral drugs, so it can only reduce the recurrence rate. Drugs for early control of genital warts
Moreover, the existing drugs for inhibiting high-risk HPV, such as Ruibeisheng, are only high-risk HPV inhibitors, and there is a lack of drugs specifically targeting low-risk HPV

Method used

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  • Application of ailanthus altissima milk gel in preparation of drugs for resisting low-risk HPV infection and preventing and treating low-risk HPV related diseases
  • Application of ailanthus altissima milk gel in preparation of drugs for resisting low-risk HPV infection and preventing and treating low-risk HPV related diseases
  • Application of ailanthus altissima milk gel in preparation of drugs for resisting low-risk HPV infection and preventing and treating low-risk HPV related diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1 HPV6 and HPV11 pseudovirus preparation

[0057] 1. Experimental method

[0058] 1. HPV6 and HPV11 pseudovirus construction

[0059] (1) Construction of structural gene expression plasmids

[0060] The codon-optimized HPV6 and HPV11 structural protein L1 and L2 genes were synthesized and cloned into the vector pCMV-MCS, and the structural gene expression plasmids were named pCMV-HPV6-L1-flag-L2-6*His and pCMV-HPV11- L1-flag-L2-6*His.

[0061] (2) Construction of plasmid

[0062] The reporter plasmids pcDNA3.1-EGFP-HPV6-E6, pcDNA3.1-EGFP-HPV6-E7, pcDNA3.1-EGFP-HPV11-E6, pcDNA3.1-EGFP-HPV11-E7 were synthesized. Hereinafter, pcDNA3.1-EGFP-HPV6-E6 / E7 means pcDNA3.1-EGFP-HPV6-E6 and pcDNA3.1-EGFP-HPV6-E7, and pcDNA3.1-EGFP-HPV11-E6 / E7 means pcDNA3.1- EGFP-HPV11-E6 and pcDNA3.1-EGFP-HPV11-E7.

[0063] (3) Routine transformation and mass extraction of plasmids

[0064]Take out 4 tubes of DH5α competent cells from the -80°C refrigerator and put them on ice. A...

Embodiment 2

[0080] Example 2 Effect of Tsubaki Milk Gel on Morphology and Proliferation of 293FT Cells

[0081] 1. Experimental method

[0082] Divide 293FT cells by 1.5×10 4 / mL density in a 96-well plate, placed at 37 ° C, 5% CO 2 Incubate overnight in the incubator. After the cells grow into a single layer of cells, wash the cells with 0.9% NaCl solution for 2 to 3 times, and dilute the mother solution (40 mg / mL) of the test product (Tsubaki milk gel) with DMEM medium containing 10% FBS for 6 Concentrations (1000, 500, 250, 125, 62.5 and 31.25 μg / mL); positive drug (podophyllin) set 6 concentrations (100, 50, 25, 12.5, 6.25 and 3.125 μmoL / L), take each group Add 100 μL of the drug into the 96-well plate, and set a blank control group at the same time, set 3 duplicate holes for each group, and place at 37°C, 5% CO 2 After 24h, 48h, and 72h, the morphology of the cells was observed under an inverted fluorescence microscope, and the cell activity after 72h was measured by the CellTite...

Embodiment 3

[0087] The impact of embodiment 3 Tsubaki milk gel on HPV6 and HPV11 pseudovirus infectivity

[0088] 1. Experimental method

[0089] Divide 293FT cells by 1.5×10 4 / mL density in a 96-well plate, placed at 37 ° C, 5% CO 2 Incubate overnight in an incubator until a monolayer of cells forms. When the cells are about 80% full, discard the culture medium and add the 100-fold TCID prepared in Example 1 50 100 μL / well of HPV6 and HPV11 pseudovirus solution. After continuing to cultivate for 2h, change to 100 μ L / hole of different concentrations of drug-containing medium (concentration is set according to cytotoxicity). , 31.25, 15.62, 7.81 and 3.90 μg / mL), the positive drug was diluted to 6 concentrations (100, 50, 25, 12.5, 6.25 and 3.125 μmoL / L), and the vehicle control group was set at the same time: simply add DMEM medium, each group 8 duplicate wells, placed at 37°C, 5% CO 2 After incubation in the incubator for 24-72 hours, observe and record the cytopathic conditions o...

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Abstract

The invention discloses application of ailanthus altissima milk gel in preparation of drugs for resisting low-risk HPV infection and preventing and treating low-risk HPV related diseases. According to the research of the invention, the ailanthus altissima milk gel can obviously reduce HPV6 and HPV11 related gene and protein expression, inhibit HPV6 and HPV11 virus proliferation, reduce virus infectivity and recover cell cycle distribution abnormality induced by low-risk HPV infection, and has a recovery treatment effect on cell development abnormality induced by low-risk HPV infection. And the Chinese toon milk gel can obviously reduce the pH value of the vagina and the virus titer of the cervical tissue after low-risk HPV infection, relieve the pathological changes of the cervical tissue and relieve the conditions of epithelial cell hyperplasia, necrosis and inflammatory cell infiltration of the cervical tissue caused by HPV6 and HPV11 infection. Therefore, the ailanthus altissima milk gel can be used for preventing low-risk HPV virus infection and treating diseases caused by low-risk HPV viruses. And the composition can play a role in the early formation stage of the condyloma acuminatum.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of Tsubaki milk gel in the preparation of medicines for resisting low-risk HPV infection and preventing and treating low-risk HPV-related diseases. Background technique [0002] Human papilloma virus (Human Papilloma Virus, HPV) is a spherical double-stranded DNA virus, and its genome sequence is about 7900bp in length. More than 160 different subtypes of HPV have been detected and are mainly transmitted to the genitals and surrounding skin through sexual behavior. [0003] Studies have reported that different HPV subtypes can cause different clinical diseases, which can be roughly divided into low-risk subtypes and high-risk subtypes according to the severity of the disease. Low-risk HPV, also known as non-cancer-related types, mainly includes HPV6, 11, 13, 42, 43, 44, 53, and 54. It can be considered that the infection of this type of HPV mainly causes comm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/06A61K36/71A61K36/65A61P31/20A61P17/12A61K31/045
CPCA61K9/06A61K36/185A61K36/489A61K36/71A61K36/65A61K36/324A61K31/045A61K47/38A61P31/20A61P17/12A61K2236/331A61K2236/39A61K2236/51A61K2236/53A61K2300/00
Inventor 吴梦瑶龚云何娟娟赵威彭冬冬张小冬姜德建
Owner ZHUZHOU QIANJIN PHARMA
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