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Method for screening drugs for targeting synthesis and assembly of bacillus tubercle cell wall core

A technology of Mycobacterium tuberculosis and cell wall, which is applied in the field of screening drugs targeting Mycobacterium tuberculosis, and can solve the problem of lack of screening drugs targeting the whole cell wall of Mycobacterium tuberculosis

Active Publication Date: 2010-12-08
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

[0004] However, the prior art still lacks a method for screening drugs targeting the whole cell wall of Mycobacterium tuberculosis, especially a method for screening drugs targeting the synthesis and assembly of the core of the cell wall of Mycobacterium tuberculosis

Method used

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  • Method for screening drugs for targeting synthesis and assembly of bacillus tubercle cell wall core
  • Method for screening drugs for targeting synthesis and assembly of bacillus tubercle cell wall core
  • Method for screening drugs for targeting synthesis and assembly of bacillus tubercle cell wall core

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] The determination of embodiment 1 inoculum size

[0072] OD 600 The value can reach 6.6-7.5, diluted to OD 600 for 3. The growth rate and final concentration of the bacteria under different culture times (8h, 10h, 12h, 14h, 24h) with different proportions of inoculum (5%, 1%, 0.5%, 0.3% and 0 / 1%) , measured by OD, the growth curve after inoculation is as follows figure 1 . Select several inoculum concentrations with small differences in absorption values ​​before culturing, and use 1 μg / ml ethambutol to evaluate them. When the inoculum amount is 0.3%, the inhibition rate difference is the most obvious, which is 56.5±5.71, while at 0.5% It was 37.4±4.22 when the inoculation amount was 1%, and 17.2±1.37 when the inoculation amount was 1%, and the inhibition of ethambutol on the bacteria in BHI medium was the most significant when the inoculation amount was 0.3%.

Embodiment 2

[0073] The determination of embodiment 2 positive control drug concentration

[0074] In BHI and BHIS medium, measure different concentration ethambutol (1ng, 10ng, 100ng, 1 μgl, 10 μg, 100 μgl, 1mg / ml) to the growth inhibitory rate of wild-type Corynebacterium glutamicum, determine the difference between the two The maximum ethambutol concentration was the concentration of the positive control drug in the screening model.

[0075] Determination of the concentration of ethambutol, the concentration of 1 μg / ml has a significant difference in the inhibition rate of wild-type Corynebacterium glutamicum in the two media, and when the concentration of 10 μg / ml is used, in the BHIS medium, sorbitol The growth of Corynebacterium glutamicum was not significantly restored. Therefore, 1 μg / ml was chosen as the measurement standard for the construction and application of drug screening models based on 96-well plates.

Embodiment 3

[0076] The feasibility verification of embodiment 3 model

[0077] Using the conditions determined above, the different effects of antibiotics with different mechanisms of action on the test strain wild-type Corynebacterium glutamicum on the two culture media were determined. For the 96-well plate method, add the known antibiotic to be tested to each well with a final concentration of 1 μg / ml. For the disk method, 50 μl of the compound at 100 μg / ml was applied to a disk with a diameter of 7 mm.

[0078] The antibiotic of table 2 different mechanism of action is to the inhibition situation of wild-type Corynebacterium glutamicum on different mediums

[0079]

[0080] a. Use concentration is 50ug / ml; b. Use concentration is 500ug / ml

[0081] In the 96-well plate-based screen, the difference in activity was represented by the difference in inhibition rate. It can be seen from Table 2 that among the drugs known to act on the cell wall of Mycobacterium tuberculosis, ethambuto...

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Abstract

The invention relates to a method for screening drugs for targeting bacillus tubercle, in particular to a method for screening drugs for targeting synthesis and assembly of a bacillus tubercle cell wall. The invention comprises the following steps of establishing a screening model for targeting the synthesis and the assembly of the bacillus tubercle cell wall core by adopting Corynebacterium glutamicum as a test organism and an BHI (Brain-Heart Infusion) and BHIS (Brain-Heart Infusion Supplemented) medium as a screening medium, and screening novel anti-tuberculous drugs and lead compounds of different enzyme systems and target spots, which act on the synthesis and assembly process of the bacillus tubercle cell wall core, from microorganism metabolic products and samples of other sources. The anti-tuberculous drugs for targeting the whole synthesis and assembly process of the bacillus tubercle cell wall core can be screened by the screening model of the invention. The model is a cell-level high-flux screening model.

Description

technical field [0001] The invention relates to a method for screening drugs targeting Mycobacterium tuberculosis, especially a method for screening drugs targeting Mycobacterium tuberculosis cell wall core synthesis and assembly, and a method for screening drugs targeting Mycobacterium tuberculosis cell wall core synthesis and assembly Screening models. Specifically, the present invention relates to using Corynebacterium glutamicum as a test bacterium, using BHI (brain-heart infusion, brain heart infusion) medium and BHIS medium as screening media, from microbial metabolites and samples from other sources Screen new anti-tuberculosis drugs and lead compounds that act on different enzyme systems and different targets in the synthesis and assembly process of the core of Mycobacterium tuberculosis cell wall. The invention also relates to the screening method and screening criteria of the screening model. Background technique [0002] The cell wall of Mycobacterium tuberculos...

Claims

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Application Information

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IPC IPC(8): C12Q1/18C12R1/32C12R1/34C12R1/15
Inventor 肖春玲高鹏郝雪秦关艳熊小椒
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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