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Brain-targeted HIV entry inhibitor polypeptide and application thereof

An entry inhibitor and brain-targeted technology, applied in the field of biomedicine, can solve problems such as viral load rebound and inability to effectively prevent HIV infection, and achieve low toxicity and strong specificity

Active Publication Date: 2021-05-28
DALI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, although HAART therapy can inhibit the replication of HIV-1 to the greatest extent, it cannot effectively prevent HIV from infecting the central nervous system.
Once the drug is stopped, the virus re-replicates and amplifies, leading to a rapid rebound in viral load

Method used

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  • Brain-targeted HIV entry inhibitor polypeptide and application thereof
  • Brain-targeted HIV entry inhibitor polypeptide and application thereof
  • Brain-targeted HIV entry inhibitor polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1AL 6 T, TL 6 Expression and purification of A

[0043] 1. Construction of recombinant plasmids

[0044] Construction of protein expression plasmids pET-28a-Ang-L6-T1144 and pET-28a-T1144-L6-Ang, Ang-L6-T1144 and T1144-L6-Ang nucleotide sequences were synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd. and approved BamHI and EcoRI were inserted into pET-28a expression vector.

[0045] Wherein, the Ang-L6-T1144 nucleotide sequence is as follows:

[0046] CGGATCC ATGACCTTTTTTTTATGGCGGCTGCCGCGGCAAACGCAACAACTTTTAAAACCGAAGAATAGCGGCGGCCGCGGCGGCACGACCTGGGAAGCATGGGACAGAGCTATTGCTGAATACGCAGCTAGGATAGAAGCTTTACTCAGAGCTTTACAAGAACAGCAAGAAAAGAATGAAGCAGCCTTAAGGGAATTA CACCACCACCACCACCACTAA GAATTC ; Wherein the underlined part contains the enzyme cleavage site and 6*his purification tag, the non-underlined part is the codon sequence of SEQ ID NO.3, the non-underlined part refers to SEQ ID NO.5, and the italicized part is the promoter.

[0047] The nucleotide sequence...

Embodiment 2HI

[0063] Example 2 Construction of HIV cell-cell fusion detection model

[0064] Studies have shown that the method of detecting HIV fusion inhibitors through cell-cell fusion models and pseudovirus models is feasible (reference 1: Li Jianbin, Chen Bin, Mi Zhiqiang, et al. Construction of human immunodeficiency virus pseudovirus drug screening model and Its application [J]. Biotechnology Communications. 2012,23(04):481-484. Reference 2: Wang Ping, Chen Huan, Luo Ronghua, et al. VSVG / HIV-1_(NL4-3)Luc Pseudovirus Screening Condition optimization and application of anti-HIV-1 drugs [J]. Chinese Pharmacology Bulletin. 2016, 32 (03): 433-438), compared with other detection models, the construction speed of these two models is fast, and it is very important for the laboratory The level requirements are low, and general laboratories can perform related experiments, and can evaluate the inhibitory effect of drugs on virus entry into cells.

[0065] Based on this, we constructed a cell-...

Embodiment 3

[0087] Example 3 The polypeptide has anti-HIV cell-cell fusion activity

[0088] References for inhibition of cell fusion experiments (Reference 4: Alam M M, Kuwata T, Tanaka K, et al.Synergistic inhibition of cell-to-cell HIV-1 infection by combinations of single chain variable fragments and fusion inhibitors[J].BiochemBiophysRep.2019 , 20:100687). Proceed as follows:

[0089] (1) On the first night, the pre-plated 293T cells were made into a cell suspension and plated in a 6-well plate.

[0090] (2) The next day, after the cells were cultured overnight and the density reached about 70-80%, transfection was performed. The pAAV-IRES-EGFP plasmid was transfected into 293T cells as a negative control, and the pAAV-EGFP-SC42 plasmid was transfected into Another well was used for fusion experiments.

[0091] (3) 12-18 hours after transfection, replace with fresh medium.

[0092] (4) On the fourth day, the cells transfected for 48 hours were observed for transfection efficiency...

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Abstract

The invention belongs to the technical field of biomedicine, and particularly relates to brain-targeted HIV entry inhibitor polypeptide and application thereof. The invention relates to a group of polypeptides which can penetrate through the blood brain barrier to target the brain, have inhibitory activity on HIV, and are further used for HIV brain infection and latent library removal. The amino acid sequence of the polypeptide is as shown in SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3 or SEQ ID NO. 4, and a derivative product of the polypeptide is also disclosed. According to the invention, candidate polypeptide drugs can be provided for prevention and treatment of human immunodeficiency virus HIV brain infection and elimination of a brain latent library.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a brain-targeted HIV entry inhibitor polypeptide and application thereof. Background technique [0002] Acquired immunodeficiency syndrome (AIDS) is a chronic infectious disease caused by human immunodeficiency virus (Human immunodeficiency virus, HIV) infection. HIV primarily attacks the human immune system, weakening the body's defenses against pathogenic infections and certain cancers. As the virus destroys immune cells, the infected person's immune system gradually becomes dysfunctional, making the body increasingly vulnerable to infection by opportunistic pathogens, cancer and certain diseases. According to differences in genes and surface antigens, HIV can be divided into two subtypes: HIV-1 and HIV-2, among which HIV-1 is the subtype of HIV widely prevalent in the world. [0003] The World Health Organization reported that at the end of 2018, there were a...

Claims

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Application Information

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IPC IPC(8): C07K14/47C12N15/12A61K38/17A61P31/18
CPCC07K14/4703A61P31/18A61K38/00
Inventor 刘奇魏雪玲石哲芳孟余王聪
Owner DALI UNIV
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