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

Vasoactive intestinal peptide I type receptor inhibitor high-throughput screening method based on fluorescence resonance energy transfer technology

A technology of vasoactive intestinal peptide and receptor inhibitors, which is applied in the field of pharmacology and can solve problems such as screening methods for less vasoactive intestinal peptide type I receptor inhibitors

Inactive Publication Date: 2016-04-06
CHINA PHARM UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are few screening methods for vasoactive intestinal peptide type I receptor inhibitors. Therefore, the establishment of convenient, fast and accurate detection methods, especially in vitro functional detection, has received more and more attention in drug screening.

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
  • Vasoactive intestinal peptide I type receptor inhibitor high-throughput screening method based on fluorescence resonance energy transfer technology
  • Vasoactive intestinal peptide I type receptor inhibitor high-throughput screening method based on fluorescence resonance energy transfer technology
  • Vasoactive intestinal peptide I type receptor inhibitor high-throughput screening method based on fluorescence resonance energy transfer technology

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach

[0016] 1. Establishment of screening method for vasoactive intestinal peptide type I receptor inhibitors

[0017] 1. Experimental materials

[0018] d2-cAMP detection kit (Cisbio, France ), VIP (Sigma-aldrich, U.S.), VIP(1-7)-GRF(8-27) (Sigma-aldrich, U.S.), DMEM, FBS (Gibco, U.S.) 384-well polypropylene microplate Cat#3573 (Corning, U.S.), disposable gun tip (Axygen, U.S.)

[0019] 2. Experimental steps

[0020] 1) Determination of the optimal concentration of vasoactive intestinal peptide type I receptor cells

[0021] 1) Different concentrations of vasoactive intestinal peptide type I receptor cells were prepared, and 5 μl was added to each well.

[0022] 2) Add 5 μl 1× reaction buffer containing VIP to each well.

[0023] 4) React at room temperature for 45 minutes.

[0024] 5) Add 10 μl × stop buffer and incubate at room temperature for 1 hour. Detect the fluorescence intensity to determine the optimal concentration of vasoactive intestinal peptide type I receptor ...

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 provides a high-throughput screening method for screening a vasoactive intestinal peptide I type receptor inhibitor; in experiments, with use of coupling of a vasoactive intestinal peptide type I receptor and a Gs protein, AC is activated, the cAMP content is increased, a product with fluorescence can be produced, the role of the vasoactive intestinal peptide type I receptor is judged by the fluorescence intensity, and thus whether the obtained compound has an inhibitory effect of the vasoactive intestinal peptide type 1 receptor is obtained. The method comprises the following steps: 1) establishment and optimization of a screening model of the vasoactive intestinal peptide I type receptor inhibitor: determining the most suitable concentration for a reaction of VIP and vasoactive intestinal peptide I type receptor cells; and 2) model reliability verification with positive drug: verifying that a positive drug IC50 is in accordance with that of a reference. The method has the advantages that the method is simple and fast, sensitivity of the fluorescence detection value is high, detection accuracy is increased, the result is stable and reliable, and the reappearance is good.

Description

technical field [0001] The invention belongs to the field of pharmacology, and uses fluorescence detection technology to construct a high-throughput screening model for vasoactive intestinal peptide type I receptor inhibitors, which is used for high-throughput screening of the inhibitory activity of test samples on vasoactive intestinal peptide type I receptors. volume detection. technical background [0002] Vasoactive intestinal peptide type I receptor is a co-receptor of neuropeptide and vasoactive intestinal peptide, mediating a variety of important biological functions, such as vasoactive intestinal peptide type I receptor mediating neuropeptide and vasoactive intestinal peptide inhibition Biological functions of appetite and anti-inflammation. Vasoactive intestinal peptide type I receptors are abundantly expressed in the brain and T cells and can regulate neuronal differentiation and T cell activation. Vasoactive intestinal peptide type I receptor, as one of the subt...

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 Applications(China)
IPC IPC(8): G01N21/64
Inventor 严明沈灵孙丹丹何玲张陆勇
Owner CHINA PHARM UNIV
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