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Compound for preventing and treating human papillomavirus infection disease and screening method of compound

A technology of human papillomavirus and screening method, which is applied in the field of compounds and their screening for the prevention and treatment of human papillomavirus infection diseases, and can solve problems such as compounds with high specificity and high inhibitors that are difficult

Active Publication Date: 2013-01-30
小江生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are no effective compounds for the prevention and treatment of human papillomaviruses at home and abroad, mainly because it is difficult to find compounds with high specificity and high Inhibitor compounds of

Method used

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  • Compound for preventing and treating human papillomavirus infection disease and screening method of compound
  • Compound for preventing and treating human papillomavirus infection disease and screening method of compound
  • Compound for preventing and treating human papillomavirus infection disease and screening method of compound

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Experimental program
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Effect test

Embodiment 1

[0053] Embodiment 1: the preparation of human papillomavirus (HPV) E1 protein and the mensuration of enzymatic activity

[0054] (1) Preparation of human papillomavirus (HPV) E1 protein

[0055] The preparation of human papillomavirus (HPV) E1 protein will take HPV35 type E1 helicase as an example. The expression and purification of HPV35E1 helicase was done by Escherichia coli expression system. The plasmid used is pGEX-2T as the vector. The condition for inducing the expression of E1 protein is 0.4mM lPTG, 16 degrees, 16 hours. Cell disruption is performed in a solution of 50mM Tris-HCl, pH 8.0 (at 4 degrees), 0.25M NaCl, 5mM EDTA, 5mM DTT, 5% glycerol, 0.2% NP-40, using a French crusher or an ultrasonic machine. The cell fragment was centrifuged at 40000x G, and the GST-E1 in the supernatant was extracted by affinity chromatography with glutathione. The GST-E1 fusion protein was cut with thrombin, and the eluted E1 protein was purified by Superdex-200 column chromatogra...

Embodiment 2

[0062] Example 2: Crystallization of Papillomavirus E1 Helicase Protein, Molecular Structure Model and Drug Screening

[0063] (1) Crystallization of papillomavirus E1 helicase protein Take bovine papillomavirus E1 as an example. The method described in Example 1 can be used for the expression, isolation and purification of the E1 protein. The crystallization of papillomavirus E1 protein is carried out at a protein concentration of 20mg / ml, in a solution containing 100mM MES (pH7.5), 300mM KPO4 (pH7.5), 190mM NaCl, 8% PEG6K, using the hanging drop gas phase diffusion method, Under the condition of 18 degrees. The crystal packet constants are: P2(1)2(1)2(1), a=133.211, b=179.192, and c=185.350, α=β=g=90. The crystal diffraction resolution was 3.1 angstroms.

[0064] (2) Molecular model of crystal structure

[0065] The overall structure of the hexamer of E1:

[0066] Such as Figure 5 (A, B) As shown, the E1 subunits assemble into a homohexameric structure with a central ...

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Abstract

The invention discloses a method for screening a compound for preventing and treating human papillomavirus (HPV) infection diseases. The method comprises the steps of adopting structure-based computer-aided drug design and screening in a mode that a human papillomavirus E1 helicase structure is used as the basis so as to obtain a candidate compound capable of suppressing the E1. The screening method has the characteristics of convenience, time saving and efficiency in Agilent hygroplasm analysis and training. The invention also discloses a compound obtained by using the screening method, such as trovafloxacin and derivatives of the trovafloxacin and gatifloxacin and derivatives of the gatifloxacin; and an inhibitor of the human papillomavirus E1 helicase structure has the characteristics of high specificity and strong pertinence, and is helpful for preventing and treating human papillomavirus infection and diseases relevant to HPV infection, such as diseases of carcinoma of uterine cervix, genital organ wart and the like.

Description

technical field [0001] The present invention relates to the field of treatment and prevention of human papillomavirus infection and cervical cancer, especially in the design and use of E1 helicase inhibitors to prevent or treat HPV-related cervical cancer and other diseases. Background technique [0002] Cervical cancer (Cervical Cancer) is second only to breast cancer in the world and threatens the life and health of women's No. 2 killer (World Health Organization data), causing more than 300,000 adult women to die of cervical cancer every year. It has been confirmed that the cause of cervical cancer is the infection of DNA carcinogenic virus of human papillomavirus (HPV). The virus is mainly transmitted through sexual contact, obligately infects human epithelial cells, especially reproductive organs, and some virus carriers show various clinical symptoms after repeated infection. After women are infected with high-risk types (HPV16, HPV18, etc.), a small number of patient...

Claims

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Application Information

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IPC IPC(8): G06F19/10A61K31/4375A61K31/496A61P35/00A61P31/20A61P15/00
Inventor 陈小江陈林李大伟楼慧强
Owner 小江生物技术有限公司
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