Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

High-throughput screening method for screening collagen transcription inhibitors for the treatment of organ fibrosis

A technology for organ fibrosis and screening methods, applied in the direction of biochemical equipment and methods, instruments, measuring devices, etc., can solve problems such as organ fibrosis and organ function decline

Active Publication Date: 2022-08-05
XIAMEN BERYL THERAPEUTICS INC
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If this repair response is excessive, strong, and out of control, it will cause organ fibrosis and lead to organ function decline

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • High-throughput screening method for screening collagen transcription inhibitors for the treatment of organ fibrosis
  • High-throughput screening method for screening collagen transcription inhibitors for the treatment of organ fibrosis
  • High-throughput screening method for screening collagen transcription inhibitors for the treatment of organ fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A high-throughput screening method for screening collagen transcription inhibitors for treating organ fibrosis, comprising the following steps:

[0025] Step 1: Clone the promoter-2000-100 region of human Col1A1, Col1A2, and Col3A1, and insert into pGL3-basic reporter gene vector (purchased from Promega) alone or in combination to obtain 4 plasmids: pGL3-Col1A1 Prom (1A1 for short), pGL3-Col1A2 Prom (1A2 for short), pGL3-Col3A1 Prom (3A1 for short) and pGL3-Col1A1 Prom -Col1A2 Prom (referred to as 1A12), the plasmid for transfection is obtained through transformation of DH5α bacteria and plasmid extraction steps;

[0026] Step 2: 3 μg of plasmid obtained in step 1 and 1 μg of internal reference pRL-TK plasmid (purchased from Promega) were transfected into NIH-3T3 cells (10 5 cells / well), 12h after transfection, 5ng / ml TGF-β1 was added to the medium for activation;

[0027] Step 3: After 24 hours of TGF-β1 activation, follow the instructions of the luciferase report...

Embodiment 2

[0029] A high-throughput screening method for screening collagen transcription inhibitors for treating organ fibrosis, comprising the following steps:

[0030] Step 1: Clone the promoter-2000-100 region of human Col1A1, Col1A2, and Col3A1, and insert into pGL3-basic reporter gene vector (purchased from Promega) alone or in combination to obtain 4 plasmids: pGL3-Col1A1 Prom (1A1 for short), pGL3-Col1A2 Prom (1A2 for short), pGL3-Col3A1 Prom (3A1 for short) and pGL3-Col1A1 Prom -Col1A2 Prom (referred to as 1A12), the plasmid for transfection is obtained through transformation of DH5α bacteria and plasmid extraction steps;

[0031]Step 2: 3 μg of the plasmid obtained in step 1 and 1 μg of the internal reference pRL-TK plasmid (purchased from Promega) were respectively transfected into human skin fibroblasts BJ (10 μg) in a 6-well plate. 5 cells / well), 5ng / ml of TGF-β1 was added to the medium after transfection for 8h, and different concentrations of TGFβR inhibitors Galuniser...

Embodiment 3

[0034] Example 3: The drug BRL-2021101 inhibits collagen transcription

[0035] The above two methods screened the compound BRL-2021101 from its own compound library. It inhibits type I collagen transcription IC50=25 μM, and the maximum potency is 94% of that of Galunisertib.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a high-throughput screening method for screening collagen transcription inhibitors for treating organ fibrosis. The pGL3-basic reporter gene vector is combined and inserted, and the plasmid for transfection is obtained by transforming DH5α bacteria and plasmid extraction; the obtained plasmid and the internal reference pRL-TK plasmid are respectively transfected into fibroblasts, and then TGF-β1 is added for activation; or at the same time TGFβR inhibitor was added; after activation, it was processed according to the luciferase reporter gene detection kit, the chemiluminescence data was read by a microplate reader, and a plasmid sensitive to TGF-β1-induced collagen synthesis was obtained. The above results confirmed this screening method practicability. Also disclosed is the application of the method in the quantitative analysis of the pharmacodynamics of drugs that inhibit I / III collagen transcription.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a high-throughput screening method for screening collagen transcription inhibitors for treating organ fibrosis. Background technique [0002] Tissue cell damage caused by any reason can lead to degeneration, necrosis and inflammatory response of tissue cells. If the damage is small, normal parenchymal cells around the damaged cells will undergo proliferative repair, which can completely restore normal structure and function. However, if the damage is larger or the repeated damage exceeds the regeneration capacity of the parenchymal cells around the injury, the extracellular matrix will proliferate to repair the defect tissue, that is, the pathological changes of fibrosis will occur. Therefore, fibrosis is essentially a repair response after tissue damage to protect the relative integrity of tissues and organs. Although the hyperplastic fibrous connective tissue repairs the defect, it...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02G01N21/64
CPCG01N33/5044G01N33/5038G01N21/6486
Inventor 王阳柏旭
Owner XIAMEN BERYL THERAPEUTICS INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com