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

A kind of 20-hydroxycholesterol-biotin molecular probe and its preparation method and application

A hydroxycholesterol and molecular probe technology, which is applied in the fields of biology, medicine and its preparation and application, can solve the problem of insufficient identification of 20-hydroxycholesterol binding protein and the like, and achieves simple and easy operation, high yield and reaction conditions. mild effect

Active Publication Date: 2018-08-24
EAST CHINA NORMAL UNIV +1
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the bipolar molecular characteristics of 20-hydroxycholesterol molecules, there has been no good method for identifying 20-hydroxycholesterol-binding proteins.

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
  • A kind of 20-hydroxycholesterol-biotin molecular probe and its preparation method and application
  • A kind of 20-hydroxycholesterol-biotin molecular probe and its preparation method and application
  • A kind of 20-hydroxycholesterol-biotin molecular probe and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1: Preparation of compound formula (1) 20-hydroxycholesterol-Biotin molecular probe

[0043] 20-hydroxycholesterol (20(S)-amine) (200mg, 0.53mmol) and Biotin-OSu (280mg, 0.8mmol) were dissolved in anhydrous DMF (10ml), triethylamine (107mg, 1.06mmol) was added, N 2 Stir at room temperature under protection for 12 hours. After the completion of the reaction as detected by TLC, add water and extract with ethyl acetate, combine the organic phases, wash with a large amount of water, wash with saturated sodium chloride solution (30 mL×3), dry over anhydrous sodium sulfate, and concentrate. Silica gel column chromatography (DCM:MeOH=10:1) gave a white solid (280 mg, 88%). 1 HNMR (500MHz, DMSO-d6) δ7.72(t, J=5.0Hz, 1H), 6.42(s, 1H), 6.35(s, 1H), 5.25(d, J=5.0Hz, 1H), 4.58( d,J=5Hz,1H),4.32–4.28(m,1H),4.15–4.10(m,1H),3.71(s,1H),3.29–3.20(m,1H),3.09(m,1H), 2.92-2.99(m,2H),2.83–2.92(m,1H),2.57(d,J=15.0 Hz,1H),2.19–2.06(m,2H),2.06–1.98(m,3H),1.93– 1.85(m,1H),1.81–1...

Embodiment 2

[0044]Example 2: Application of 20-hydroxycholesterol-Biotin molecular probe in identifying 20-hydroxycholesterol-binding protein.

[0045] Smoothened proteins can bind 20-hydroxycholesterol molecules at the cysteine-rich region of their N-terminus. First, at 4°C, the 20-hydroxycholesterol-Biotin molecular probe prepared in Example 1 was incubated with Avidin beads for 4 hours, and the 20-hydroxycholesterol-Biotin molecular probe was coated on the Avidin beads . Then the free 20-hydroxycholesterol-Biotin molecular probe was washed to obtain Avidin beads coated with 20-hydroxycholesterol-Biotin molecular probe. After the Smoothened protein was transfected in 293T cells, cultured at 37°C for 48 hours, the cells were collected, and treated with 10 mM HEPES pH=7.5, 300 mM NaCl and 1.3% (mass / volume) n-dodecyl-β-d-maltopyranoside (DDM , Anatrace) to lyse the cells, use Flag beads to specifically immunoprecipitate the Smoothened protein, 4 degrees Celsius for 3 hours, then use 150...

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 20-hydroxycholesterol-Biotin molecular probe and a preparation method and application thereof. 20-hydroxycholesterol is used as a raw material and performs condensation reaction with Biotin-OSu to obtain the 20-hydroxycholesterol-Biotin molecular probe shown as a formula (1). The preparation method has the advantages of mild reaction conditions, simple and easy operation and high yield. The invention further discloses application of the 20-hydroxycholesterol-Biotin molecular probe in identification of 20-hydroxycholesterol binding proteins.

Description

technical field [0001] The invention belongs to the technical field of biology, medicine and its preparation and application, and specifically relates to a 20-hydroxycholesterol-Biotin molecular probe, its preparation, and its application in the process of identifying 20-hydroxycholesterol-binding protein. Background technique [0002] As an important cholesterol derivative, 20-hydroxycholesterol can activate the hedgehog pathway by binding smoothened protein. Abnormal activation of the hedgehog pathway often leads to tumors and other diseases, while abnormal silencing can lead to developmental disorders and other diseases. The study of the combination of 20-hydroxycholesterol and smoothened protein is of great significance to the study of diseases related to the hedgehog pathway, and the identification of new 20-hydroxycholesterol-binding proteins also has important scientific research and clinical value. However, due to the bipolar molecular characteristics of 20-hydroxyc...

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): C07J9/00G01N33/92G01N33/68
CPCC07J9/00G01N33/6803G01N33/92
Inventor 仇文卫宋保亮杨连芳唐静洁肖旭肖洁心赵青玲
Owner EAST CHINA NORMAL 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