A catalyst-free hydrazone-linked polypeptide or protein chemical modification method based on electron-deficient benzaldehyde

A technology for protein modification and linking peptides, applied in the field of chemical modification of peptides and/or proteins, can solve the problems of limited application scope and not many, and achieve the effect of simple feeding method

Active Publication Date: 2021-10-01
UNIV OF SCI & TECH OF CHINA
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Traditional hydrazone ligation reactions have so far been limited in scope due to the need for the addition of toxic catalysts
Although there are many new methods for hydrazone linkage reaction without catalyst addition, there are not many methods that can be used for peptide and / or protein modification conveniently and universally. Some methods only realize the reaction at the model molecular level, while others can only Simple fluorescent molecular modification of peptides and 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 catalyst-free hydrazone-linked polypeptide or protein chemical modification method based on electron-deficient benzaldehyde
  • A catalyst-free hydrazone-linked polypeptide or protein chemical modification method based on electron-deficient benzaldehyde
  • A catalyst-free hydrazone-linked polypeptide or protein chemical modification method based on electron-deficient benzaldehyde

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] The electrophile shown in embodiment 1 preparation formula 1, 2 and 3

[0104] (1) The electrophile of formula 1 is prepared by chemical synthesis. The specific process is as follows:

[0105]

[0106] Commercially available 4-amino-2-chlorobenzonitrile (1.5 g, 10 mmol, purchased from TCI (Shanghai) Chemical Industry Development Co., Ltd.) was weighed and added to a 25 mL round bottom flask. Add 5mL of deionized water and 12mmol of concentrated hydrochloric acid to it. It should be noted that the concentrated hydrochloric acid needs to be added dropwise slowly. After the dropwise addition was complete, stirring was continued for 30 minutes. Sodium nitrite (0.76 g, 11 mmol) was then added to the flask and the reaction was stirred until the precipitate mostly disappeared (10 minutes or so). After observing this phenomenon, potassium iodide (1.82 g, 11 mmol) was added to the flask, and the stirring reaction was continued for 1 hour. After the reaction was completed,...

Embodiment 2

[0118] Embodiment 2 prepares the polypeptide hydrazone connection product of formula 9

[0119] The reaction formula of this embodiment is as follows:

[0120]

[0121] Concrete reaction steps are as follows:

[0122] (1) Dissolve the powder of polypeptide hydrazide (Leu-Tyr-Arg-Ala-Phe-NHNH2) in PME buffer (100mM PIPES, 1mM MgSO 4 , 2mM EGTA, pH 7.0) and prepared as a 1 mg / mL stock solution, the electron-deficient benzaldehyde compound 1 was dissolved in ethanol and prepared as a 10 mM stock solution.

[0123] (2) Take a 4mL plastic round bottom centrifuge tube and add 1mL of PME buffer into the tube. Then take an appropriate amount of the polypeptide hydrazide stock solution and the electron-deficient benzaldehyde 1 stock solution prepared in step (1) into the above-mentioned centrifuge tube, and the final concentrations of the two are 30 μM and 200 μM, respectively. Add a magnetic stir bar to the centrifuge tube and stir the reaction at room temperature for 30 minutes...

Embodiment 3

[0132] Embodiment 3 prepares the protein hydrazone connection product of formula 12

[0133] The reaction formula of this embodiment is as follows:

[0134]

[0135] This example can prove that the present invention is suitable for coupling between proteins and drug molecules, and may be applied in the field of antibody-drug coupling in the future.

[0136] Concrete reaction steps are as follows:

[0137] (1) Dissolve the powder of ubiquitin hydrazide in PME buffer (100mM PIPES, 1mMMgSO 4 , 2mM EGTA, pH 7.0) and prepared as a stock solution of 1mg / mL, the electron-deficient benzaldehyde (formula 3) coupled with dasatinib molecules was dissolved in N,N-dimethylformamide and prepared as 10 mg / mL stock solution.

[0138] (2) Take a 4mL plastic round bottom centrifuge tube and add 1mL of PME buffer into the tube. Then take an appropriate amount of the protein hydrazide stock solution prepared in step (1) and the electron-deficient benzaldehyde stock solution coupled with da...

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 method for modifying a hydrazone-linked polypeptide or protein without adding a catalyst, the method comprising the following steps: under the condition of an aqueous buffer, using an electron-deficient benzaldehyde compound represented by formula III as the hydrophilic The electroreagent uses the hydrazide of the polypeptide or protein shown in the formula II as a nucleophile to carry out the hydrazone linking reaction to obtain a hydrazone linking product having the structure shown in the general formula I, wherein, in the formula III, R is a variable structure Substituents, for example, methyl groups, fluorescent molecule substituents, drug molecule substituents, etc. The present invention also provides the electron-deficient benzaldehyde compound of formula III and the polypeptide or protein modified by the electron-deficient benzaldehyde compound obtained by the hydrazone linking polypeptide or protein modification method.

Description

technical field [0001] The invention belongs to the technical field of chemical modification of polypeptides and / or proteins. Specifically, the invention relates to a hydrazone-linked polypeptide or protein chemical modification method based on electron-deficient benzaldehyde compounds without catalyst addition. Background technique [0002] The chemical modification of peptides and proteins has been an extremely important part of the field of chemical biology since its inception (Angew. Chem. Int. Ed. 2009, 48, 6974.). Due to its good bio-orthogonality, hydrazone ligation reaction has been widely used in the field of chemical modification of peptides and proteins. At present, there are mainly the following methods for the modification of polypeptides and proteins using the hydrazone linkage method: [0003] (1) Hydrazone linkage reaction catalyzed by aniline or aniline catalyst. Polypeptide and protein modification reactions need to be carried out under neutral conditions...

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): C07K1/107C07K1/13A61K47/64C07C69/716C07D417/12C07D271/12
CPCA61K47/64C07C69/716C07D271/12C07D417/12C07K1/1077C07K1/13
Inventor 许杨石景王玉
Owner UNIV OF SCI & TECH OF CHINA
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