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A kind of Streptomyces producing anti-HIV active substance and its application

A streptomyces, HIV-1 technology, applied in the direction of medical preparations containing active ingredients, bacteria, antiviral agents, etc., can solve problems such as drug resistance and rapid virus mutation

Active Publication Date: 2020-10-27
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The pathogenic virus is naturally highly mutated, and under the selection pressure of drugs, it is easy to cause rapid mutation of the virus and thus drug resistance.

Method used

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  • A kind of Streptomyces producing anti-HIV active substance and its application
  • A kind of Streptomyces producing anti-HIV active substance and its application
  • A kind of Streptomyces producing anti-HIV active substance and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1 Streptomyces (Streptomyces sp.) CPCC 201585 prepares the method for Surfactin compounds

[0057] Streptomyces sp. of the present invention (Streptomyces sp.) CPCC 201585, which is isolated from the soil sample of Menghai County, Jinghong, Yunnan Province, has been preserved in the General Microorganism Center (CGMCC) of China Committee for Microorganism Culture Collection on December 30, 2014, The address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, the postal code is 100101, and the deposit number is CGMCCNo.10265.

[0058] Such as figure 1 Shown, utilize streptomyces (Streptomyces sp.) CPCC 201585 to prepare the method for Surfactin compounds, the steps are as follows:

[0059] 1), the fermentation of the Streptomyces sp. CPCC 201585

[0060] Strain activation: prepare slant culture medium, divide into test tubes, sterilize at 121°C for 30 minutes, and then make slant; transfer the preserved Streptomyces sp. sky;

[0061] The formu...

Embodiment 2

[0072] Anti-HIV activity test of embodiment 2 Surfactin compounds 1-4

[0073] The anti-HIV-1 activity of the active compounds was determined by the pseudovirus cellular level pharmacological screening model.

[0074] 1. Principle of pharmacological screening model

[0075] This high-throughput screening model uses recombinant technology to co-transfect 293T cells with plasmids expressing HIV-1 core and VSV-G to generate VSVG / HIV pseudoviral particles with VSV-G shell wrapping HIV-1 core. -1 replicates as a cellular level screening model for targets.

[0076] The HIV-1 genome has removed the env of HIV-1, so the virus particles formed after its separate transfection cannot infect the host cell again, and can be operated in a conventional laboratory; at the same time, a luciferase reporter gene is introduced at the position of the nef gene, The activity of the reporter gene can reflect the replication level of the pseudovirus in the cell. The receptors of vesicular stomatiti...

Embodiment 3

[0090] Example 3 Surfactin compounds 1-4 inhibit Vif from degrading APOBEC3G (hA3G)

[0091] 1. The obtained Surfactin compounds 1-4 inhibit Vif from degrading hA3G

[0092] 293T was plated on a 6-well plate with a cell concentration of 2.0×105 cells / ml. After 24 hours, hA3G (600ng) and vif (200ng) were co-transfected with Lipofectamine 2000 (Invitrogen) as the transfection reagent. After 12 hours of transfection, add the positive control drug MG132 ((5 μM)) and the obtained Surfactin compounds 1-4 (each 20 μM), and act for 36 hours, collect the cell samples, add the loading buffer and heat in a metal bath, and store at -20 ℃.

[0093] After the test samples were subjected to polyacrylamide electrophoresis and membrane transfer, antibodies were added in sequence. Primary anti-HA antibody (hA3G) 1:3000, vif antibody 1:5000, secondary antibody goat anti-rabbit 1:3000. As a result, it was found that the positive control drug i.e. MG132 and the screened Surfactin compounds 1-4...

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Abstract

The present invention relates to Streptomyces sp. CPCC 201585 and applications thereof. According to the present invention, the acetone extract of the Streptomyces sp. CPCC 201585 fermented broth can significantly inhibit the replication of HIV-1, and the Surfactin compound isolated from the mycelium extract can provide the HIV-1 replication inhibition effect by inhibiting the degradation of Vif on hA3G.

Description

technical field [0001] The invention belongs to the field of microbial pharmacy, and in particular relates to a Streptomyces sp. CPCC 201585 strain producing anti-HIV-1 active substances and its application. Background technique [0002] HIV (Human Immunodeficiency Virus) virus, the full name of human immunodeficiency virus in Chinese, is a kind of retrovirus. After the HIV virus invades the host cells, it will seriously damage the immune function of the host body, thereby causing severe opportunistic infections and causing secondary immunodeficiency syndrome characterized by malignant diseases, which is acquired immunodeficiency syndrome (acquired immunodeficiency syndrome, AIDS). The first case of AIDS was discovered in the United States in 1981, and then began to spread around the world, seriously threatening human health. HIV virus can be divided into HIV-1 type and HIV-2 type, and the genomes of the two have great differences, among which HIV-1 is the main pathogen th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/74A61K38/08A61P31/18C12P21/02
Inventor 余利岩庞旭岑山马铃方晓梅魏玉珍李晓宇刘红宇张玉琴
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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