Application of Retro-2cycl and related derivatives

A retro-2.1, derivative technology, applied in the field of new drug use

Active Publication Date: 2017-06-23
CHANGCHUN BCHT BIOTECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently about Retro-2 cycl and Retro-2.1 and related derivatives have no reports on the inhibitory effect of picornavirus infection

Method used

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  • Application of Retro-2cycl and related derivatives
  • Application of Retro-2cycl and related derivatives
  • Application of Retro-2cycl and related derivatives

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Example 1: Construction of 293S cells

[0100] The synthetic EV71 receptor human SCARB2 gene fragment (GenBank accession no: NM_005506.3, synthesized by Shanghai Jierui Company) with EcoR I & Xba I restriction sites at both ends was inserted into the lentiviral vector Lenti-XpLVX-Puro vector (purchased from Clontech), and use the transfection reagent in the lentiviral vector kit (purchased from Clontech) to transfect human embryonic kidney cells (HEK 293 cells, purchased from ATCC, accession number #CRL-1573) according to the instructions to obtain stable transfection human SCARB2 HEK 293 cells, referred to as 293S cells.

Embodiment 2

[0101] Example 2: Retro-2 cycl and Retro-2.1 cytotoxicity and inhibition of cytopathic effects caused by enterovirus 71

[0102] Take 293S cells in the logarithmic growth phase, and use 3×10 4 The density of cells / well was inoculated in 96-well culture plate and cultured for 24 hours. 293S cells at 37°C and 5% CO 2 The culture medium used was DMEM cell culture medium containing 1.25 μg / ml puromycin (purchased from Clontech) supplemented with 10% fetal bovine serum (FBS) (abbreviated as 10% FBS-DMEM, both FBS and DMEM were purchased from Sigma).

[0103] In detection of Retro-2 cycl When the cytotoxicity of Retro-2.1 and Retro-2.1 was detected, the original cell culture medium was sucked off, and 100 μl of Retro-2 containing different concentrations were added to each well. cycl and Retro-2.1 supplemented with 2% fetal bovine serum DMEM cell culture fluid (abbreviated as 2% FBS-DMEM), so that Retro-2 cycl And the final concentration of Retro-2.1 is 500μM, 250μM, 125μM, 62...

Embodiment 3

[0124] Example 3: Retro-2 cycl Inhibition of viral plaque effect caused by enterovirus 71 and Retro-2.1

[0125] Take 293S cells in the logarithmic growth phase, and use 5×10 5 The density of cells / well was inoculated in 12-well culture plate, and cultivated for 24 hours. 293S cells at 37°C and 5% CO 2 Next, the culture medium used was 10% FBS-DMEM containing 1.25 μg / ml puromycin. Aspirate the original cell culture solution from each well, and before adding 500 μl EV71 to infect (the infection volume is 100PFU / well), add 500 μl of Retro-2 containing different concentrations to the cells in each well cycl and Retro-2.1 in 2% FBS-DMEM cell culture medium to make Retro-2 cycl The final concentrations of Retro-2.1 and Retro-2.1 were 25 μM, 12.5 μM, 6.25 μM, 3.13 μM, 1.56 μM and 6.25 μM, 1.56 μM, 0.39 μM, 0.098 μM, 0.024 μM, and each concentration was repeated for 5 hours; Add EV71 to infect and maintain the presence of the drug, continue to culture the cells for 1 hour, suck ...

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PUM

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Abstract

The invention provides an application of Retro-2cycl and/or Retro-2.1 and related derivatives in preparing drugs for preventing and/or treating diseases induced by small RNA virus infection. Researches in the invention verify that the Retro-2cycl and/or Retro-2.1 and the related derivatives can effectively inhibit cytopathic effect and viral plaque effect induced by an enterovirus 71, remarkably reduce the output of progeny viruses of the enterovirus 71 and remarkably inhibit an enterovirus 71 infected mouse. The researches prove that Retro-2cycl and/or Retro-2.1 and related derivatives can effectively prevent and/or treat diseases induced by small RNA virus infection.

Description

technical field [0001] The present invention relates to the field of new application of medicine, in particular to Retro-2 cycl And / or the application of Retro-2.1 and related derivatives in the preparation of drugs for preventing and / or treating diseases caused by picornavirus infection. Background technique [0002] Picornaviruses are tiny, non-enveloped, positive-strand RNA viruses that currently contain 17 virus genera, such as enterovirus, rhinovirus, and hepatovirus. Picornaviruses are the pathogens of many human and animal diseases, and the more common pathogenic viruses include Enterovirus 71 (EV71), Coxsackievirus (Coxsackievirus), Poliovirus (Poliovirus) and Hepatitis A virus ( Hepatitis A virus), etc., can cause various diseases such as hand, foot and mouth disease, poliomyelitis, hepatitis A, etc., leading to respiratory inflammation, hand, foot and mouth inflammation, meningitis, acute myelitis, cardiovascular disease, hemorrhagic conjunctiva Inflammation, hep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/517A61P31/14
CPCA61K31/517
Inventor 苏维恒姜春来孔维代文文丹尼尔·吉莱让-克里斯托夫·桑塔于廉·巴比尔吴瑜
Owner CHANGCHUN BCHT BIOTECH
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