Compound containing disulfide bond and its use and preparation method

A compound and disulfide bond technology, which is applied in the field of compounds containing disulfide bonds, can solve the problems of difficult separation of detergents and extracted substances, and achieve the effect of good film solubility and protein stabilization

Active Publication Date: 2021-08-24
SHANGHAI TECH UNIV
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the above-mentioned shortcoming of prior art, the object of the present invention is to provide a kind of compound containing disulfide bond, is used to solve the difficult problem of separation of detergent and extracted substance in prior art; Optimized membrane protein Preparation and Functional Studies

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
  • Compound containing disulfide bond and its use and preparation method
  • Compound containing disulfide bond and its use and preparation method
  • Compound containing disulfide bond and its use and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Disulfide bond bridged cleavable detergent molecule and its synthesis

[0063] 1. Disulfide bridged detergents, the structural formulas of which are the above-mentioned (XII)-(XV).

[0064] 2. The synthesis method is as follows, reagents and reaction conditions: i) sodium thiobenzenesulfonate, tetrabutylammonium bromide, acetonitrile, 70°C; ii) triethylamine, dichloromethane, 0°C to room temperature; iii ) sodium methoxide, methanol, room temperature.

[0065]

[0066] Specific steps are as follows:

[0067] (i) Dissolve 14g of alpha-D-bromomaltose heptaacetate 1 in 20ml of anhydrous acetonitrile, add 644mg of tetrabutylammonium bromide (Titan, product number: G28296B) and 8g of sodium thiobenzenesulfonate (Datang Medicine, Cat. No.: BL-06097). The reaction solution was refluxed at 70° C. for 6 hours (or reacted overnight). After the reaction was completed, the solvent was removed by rotary evaporation. Add 100mL ethyl acetate and 100mL water, separate...

Embodiment 2

[0084] Example 2 Evaluate the micelle properties formed by each detergent in Example 1

[0085] 1. CMC determination

[0086] The critical micelle concentration (CMC) of each detergent was determined by fluorescent dye method. The disulfide bond bridged detergents with structural formulas (XII)-(XV) are formulated into a gradient solution, and the CMC value of the detergent is determined according to the mutation point of the fluorescence curve of the hydrophobic fluorescent dye. For example, prepare concentrations of 0, 42, 84, 126, 168, 204, 240, 360, 480 with double distilled water containing 40uM fluorescent dye 8-amino-1-sulfonic acid naphthalene ammonium salt (Baringwei, product number: 442848). , 600, 800 and 1000uM detergent solutions and incubate for 20 minutes in the dark. Measure the fluorescence curve at 477nm / 388nm, and the abrupt change point of the curve is the CMC value of the detergent.

[0087] 2. Dynamic Light Scattering Experiment

[0088] Structural fo...

Embodiment 3

[0093] Example 3 Evaluation of CCD-2

[0094] The detergent CCD-2 in Example 1 was used to dissolve and purify GPCRs of different families, specifically including adenosine receptor 2A subtype protein (A2a protein, class A GPCR), glucagon-like peptide-1 receptor protein (GLP-1R, class B GPCR) and Smoothened receptor (SMO, class F GPCR).

[0095] 1. Specific steps:

[0096] Expression of GPCRs (A2a, GLP-1R, and SMO) in SF9 cells (invitrogen, Cat. No. 11496-015):

[0097] Cells were cultured at 37°C until the cell density reached 1.0-1.3x10 6 cells / mL were collected and broken to obtain cell membranes. Wash the cell membrane with low-salt buffer (10mM magnesium chloride (Sigma-Aldrich, product number M4880), 20mM potassium chloride (Sigma-Aldrich, P9541), 20mM Hepes (ABCONE, product number H33755) at pH 7.5) (50mL*3 times) and centrifuged, and then washed with high-salt buffer (1M sodium chloride (Shanghai Sangon, product number A610476), 10mM magnesium chloride, 20mM potass...

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 compound containing a disulfide bond and its use and preparation method. The general structural formula of the compound is: . Wherein, said A is a substituted or unsubstituted hydrophilic monosaccharide group, oligosaccharide group, or ammonium ammonium salt group, and said B is a hydrophobic aliphatic group within 40 carbon atoms. In the present invention, by introducing a disulfide bond at the corresponding position of the small molecule, it not only retains the film dissolution and protein stabilization effect equivalent to or even better than that of the original detergent, but also can completely realize the In situ degradation, degraded into two thiol fragments, losing the properties of surfactants. With the aid of the compound, it can be used to prepare and study membrane proteins.

Description

technical field [0001] The invention relates to a compound containing a disulfide bond, its use and preparation method. Background technique [0002] Detergents, also known as surfactants, are amphiphilic molecules with both hydrophilic polar groups and hydrophobic non-polar groups. One end is a hydrophilic group, and the other end is a hydrophobic group; the hydrophilic group is often a polar group, such as carboxylic acid, sulfonic acid, sulfuric acid, amino group or amine group and its salt, hydroxyl group, amide group, ether bond etc. can also be used as polar hydrophilic groups; while hydrophobic groups are often non-polar hydrocarbon chains, such as hydrocarbon chains with more than 8 carbon atoms. According to its dissociation state characteristics in water, it is generally divided into ionic detergents (including anionic and cationic), nonionic detergents and zwitterionic detergents. [0003] In the study of membrane proteins, detergents are used to compete and wea...

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): C07H5/10C07H1/00C07K1/14
CPCC07H1/00C07H5/10C07K1/14
Inventor 陶厚朝薛东香
Owner SHANGHAI TECH UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap