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Polypeptide as well as preparation method and application thereof in inhibition of HIV

A lentivirus and amino acid technology, applied in the field of biomedicine, can solve the problems of being unable to clear HIV latent pools to cure AIDS, unable to kill HIV-infected cells, and achieve the effect of less drug-resistant strains

Active Publication Date: 2019-03-29
SHANXI JINBO BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

More importantly, the proposed protein inactivators and broad-spectrum neutralizing antibodies can only neutralize HIV, but still cannot kill HIV-infected cells, especially HIV latently infected cells, so they cannot remove HIV latent pools and cure AIDS

Method used

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  • Polypeptide as well as preparation method and application thereof in inhibition of HIV
  • Polypeptide as well as preparation method and application thereof in inhibition of HIV
  • Polypeptide as well as preparation method and application thereof in inhibition of HIV

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Embodiment 1: Binding experiment of the polypeptide of the present invention

[0097] 1.1 Experimental materials and methods

[0098]Based on enzyme-linked immunosorbent assay (ELISA), it was detected whether the F9170 polypeptide (SEQ ID NO: 2) (synthesized by Synpeptide Co., Ltd.) could interact with the following HIV envelope protein-derived proteins or polypeptides (all Synpeptide Co., Ltd. Synthetic) combination: gp120 (SEQ ID NO:19): HIV envelope protein gp120 subunit; gp140 (SEQ ID NO:20): HIV envelope protein removes the transmembrane domain and intracellular sequence part; N63 (SEQ ID NO:21): NHR domain on HIV envelope protein subunit gp41; C34 (SEQ ID NO:22): CHR domain on HIV envelope protein subunit gp41; MPER (SEQ ID NO:23): HIV envelope Proximal membrane domain on protein subunit gp41; LLP1(SEQ ID NO:24) / LLP2(SEQ ID NO:25) / LLP3(SEQ ID NO:26): HIV envelope protein subunit gp41 intracellular segment LLP1 / LLP2 / LLP3 domain; LLP1-RH (SEQ ID NO: 27): 828-842 ...

Embodiment 2

[0102] Embodiment 2: Polypeptide of the present invention inactivates HIV virus

[0103] 2.1 Experimental materials and methods

[0104] Detection of F9170 polypeptide (SEQ ID NO:2), F9170 scrambled polypeptide (F9-scr) (SEQ ID NO:30) and T20 (SEQ ID NO:18) (all synthesized by Synpeptide Co., Ltd.) to inactivate HIV Virus strain HIV-1IIIB, Bal, TZA68 / 125A and 92TH009 virus strain (from NIH AIDS Reagent Program, product number is 398,510,11255,1656 respectively) The method of ability is as follows: each 50 μ L above-mentioned each HIV virus (with serum-free 1640 medium (Meilunbio, product number MA0215, the 1640 medium used in all embodiments is this product) dilution, the final concentration is 200 times TCID 50 ) and 50 μL diluted (4 times) F9170 polypeptide, F9170 scrambled polypeptide and T20 (diluted with serum-free 1640 medium, initial concentration is 5 μM) and incubated at 4°C for 1 hour. Add PEG6000 (Sigma-Aldrich, Cat. No. 1546580) to make the final concentration 3%...

Embodiment 3

[0108] Example 3. F9170 specifically inhibits HIV pseudoviruses.

[0109] 3.1 Experimental materials and methods

[0110] Pseudovirus packaging: HIV envelope protein particles or MERS-CoV envelope protein particles (HIV envelope protein particles are from AIDS Reagent Program, item number is 324; MERS-CoV envelope protein particles are from cooperative research group New York Blood Center Viral Immunology research group) and HIV backbone plasmid (pNL4-3.Luc.R-E, from NIH AIDS ReagentProgram, item number: 3418) were used to package HIV and MERS-CoV pseudoviruses. Two kinds of plasmid HIV envelope protein particles or MERS-CoV envelope protein particles and HIV backbone plasmid (5 μg DNA melted in 100 μl normal saline) were simultaneously used VigoFect reagent (Vigolas Biotechnology (Beijing) Co., Ltd., catalog number T001) ( 2 μl of reagent diluted to 100 μl of saline) were transfected into 293T cells (purchased from ATCC) according to the manufacturer's instructions (200,000-...

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Abstract

The invention provides polypeptide as well as a preparation method and an application thereof in inhabitation of HIV. The polypeptide comprises an amino acid sequence shown in X-W / A-E / A-A / E-L / A-X-Y / A-L / A-W / A-N / A-L / A-L / A-Q / A-Y / A-W / A. The invention also provides nucleic acid coding the polypeptide, a carrier containing the nucleic acid, a host, a preparation or use method and an application. The invention further provides fusion protein, a pharmaceutical composition or conjugated containing the polypeptide.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a therapeutic agent for inactivating HIV. Background technique [0002] According to the data provided by the United Nations AIDS Program (UNAIDS), since it was discovered in the 1980s, the human immunodeficiency virus (HIV) has caused a total of more than 70 million people to be infected and more than 30 million people to die. Now, There are still about two million new infections each year. AIDS is still a major threat in the field of human public health because there is still no cure drug and vaccine available so far. Effective treatment has always been a focus of research in this field. The emergence of Highly Active Combined Antiretroviral Therapy (HAART) has greatly reduced the morbidity and death of AIDS among HIV-infected patients. However, the current clinically used drugs still have disadvantages such as strong toxic side effects and easy induction of drug-resistant virus st...

Claims

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Application Information

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IPC IPC(8): C07K14/16C12N15/49C12N15/62C12N15/63A61K38/10A61P31/18
CPCA61K38/00A61P31/18C07K14/005C07K2319/00Y02P20/55
Inventor 陆路姜世勃王茜苏珊杨霞
Owner SHANXI JINBO BIO PHARMA CO LTD
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