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Polypeptide of controlling IIIv virus fusion and its use

A technology of virus envelope, -X1-X2WN-X3-X4-TWMEWER-X5-IE-X6-YTKLIY-X7-IL-X8-SQEX9-, applied in the field of polypeptides that inhibit virus fusion, can solve the problem of large molecular weight, Difficulties in synthesis, etc.

Inactive Publication Date: 2007-05-02
INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0020] Although T-20, T-1249, TR-290999 and TR-291144 are currently the most effective polypeptide HIV fusion inhibitors, their common disadvantage is that the molecular weight is relatively large compared to other anti-HIV drugs, so the synthesis is relatively difficult

Method used

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  • Polypeptide of controlling IIIv virus fusion and its use
  • Polypeptide of controlling IIIv virus fusion and its use
  • Polypeptide of controlling IIIv virus fusion and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] Synthesis and purification of embodiment 1 polypeptide

[0111] The polypeptide provided by the present invention can be synthesized on an ABI 433 solid-phase synthesizer of Applied Systems Biosystems in the United States, and the modification of the polypeptide is done manually. The amino acid used in the synthesis is protected with Fmoc (product of Advanced Chemtech, USA), and the resin used is Rink resin (product of Advanced Chemtech, USA). During synthesis, 1-hydroxybenzotriazole (HoBt) (product of Advanced Chemtech, USA) was dissolved in N-methylpyrrolidone (NMP) (PE company) as an activator, and dicyclohexylcarbodiimide (DCC) was used (Acros Company) was used as a coupling agent, and piperidine (Piperidine) (Shanghai Jier Biochemical) was used to remove the protecting group. Amino acids all have an L-type chemical structure, and they are sequentially coupled to the Rink resin.

[0112]The usage amount of Rink resin and the usage amount of Fmc protected amino aci...

Embodiment 2

[0124] The CD spectrum determination of embodiment 2 polypeptide secondary structure

[0125] 1. Purpose of the experiment

[0126] By measuring the changes in the secondary structure (such as helical content) of the polypeptide provided by the invention and N36 (a sequence from the NHR region of HIV gp41, containing the action site of the polypeptide of the invention and T20, etc.) and their complexes To analyze the intermolecular interaction between the polypeptide of the present invention and N36. The stronger the interaction, the higher the activity, and vice versa.

[0127] 2. Experimental instruments, reagents and methods

[0128] Instrument: JASCO J-715-150L type

[0129] Parameter selection: resolution 0.1nm, wave width 5.0nm, response time 4.0s, wavelength 200-250nm

[0130] Dissolving buffer: 50mM NaH 2 PO 4 (Containing 150mM NaCl)pH=7.2

[0131] Peptide concentration: 10μM

[0132] The secondary structure determination of 18 polypeptides provided by the inve...

Embodiment 3

[0141] Example 3 Gel HPLC detection of the binding effect of the polypeptide of the present invention and N36

[0142] 1. Experimental purpose and principle

[0143] The purpose is to prove whether there is a strong interaction between the two by detecting whether the polypeptide of the present invention binds to N36.

[0144] In a gel column, substances are separated according to their molecular weights, and substances with larger molecular weights are eluted first. In this experiment, if the polypeptide forms a complex with N36, it will be eluted first after the molecular weight increases. If the complex cannot be formed, the mixture will form two peaks of equal height, corresponding to the polypeptide and N36 provided by the present invention respectively.

[0145] 2. Experimental instruments, reagents and methods

[0146] Gel column: TSK-G3000SWxl, 5μm, 7.8mm×300mm (product of Japan TOSOH company)

[0147] Instrument: Bio-Rad 13507 pump, Bio-Dimension TM UV / VIS detect...

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Abstract

This invention provides a category of polypeptide that restrain HIV virus to mix together. According to the mechanism of action of HIV virus and adaptor molecule which is on the surface of cellular membrane in the process of membrane fusion, this invention designs an active polypeptide which can bind with virus surface glycoprotein gp41. Effect situs of mentioned polypeptide is at trimer NHR lateral 'long point' and bilateralis that is in gp41 molecule, it can effectively restrain reproduction of HIV virus. The polypeptide of this invention compared with present medicine, has small molecular weight, high activity, good water solubility. Using polypeptide that is mentioned by this invention can prepare medicine which can restrain many kinds of viral infection.

Description

Technical field: [0001] The invention relates to a class of polypeptides for inhibiting virus fusion, in particular to a class of polypeptides for inhibiting HIV virus fusion. The present invention also relates to the use of said polypeptide. Background technique: [0002] Some viruses with envelopes, such as human immunodeficiency virus (Human Immunodeficiency Virus, HIV), human respiratory syncytial virus and hepatitis B virus, the whole life cycle can be divided into four parts: virus and cell membrane fusion, genetic material enters the cell ; reverse transcription of viral DNA and integration into host cell chromosomes; transcription, translation, modification of viral proteins; assembly and budding of virus particles. What these viruses have in common is that there is a process of membrane fusion in the life cycle, and the structures of the proteins involved in the fusion process are similar. [0003] Taking the drugs for the treatment of HIV infection as an example,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/005C07K14/155C07K19/00A61K38/16A61P31/12
Inventor 戴秋云成健伟何玉先董铭心何俊明余练康
Owner INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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