Bio-orthogonal activated time-resolved response type rare earth probe and its preparation method and application

A time-resolved, bio-orthogonal technology, applied in the direction of material excitation analysis, chemical instruments and methods, preparations for in vivo experiments, etc., can solve the problems of high copper ion toxicity, unfavorable large-scale promotion, large fluorescence background, etc., to achieve good water solubility

Active Publication Date: 2021-08-06
SOUTH CHINA NORMAL UNIVERSITY
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

【a) O.T. Keppler, R. Horstkorte, M. Pawlita, C. Schmidt, W. Reutter, Glycobiology 2001, 11, 11R–18R; b) D.H. Dube, C.R. Bertozzi, Curr. Opin. Chem. Biol. 2003, 7, 616 –625; c) T.-L. Hsu, S.R. Hanson, K. Kishikawa, S.K. Wang, M. Sawa, C.H. Wong, Proc. Natl. Acad. Sci. U.S.A. 2007, 104, 2614–2619; d) J. Du, M.A. Meledeo, Z. Wang, H.S. Khanna, V.D. Paruchuri, K.J. Yarema, Glycobiology 2009, 19, 1382–1401; e) S. Stairs, A.A. Neves, H. Y.A.Wainman, H.Ireland-Zecchini, K.M.Brindle, F.J.Leeper, ChemBioChem 2013,14,1063–1067.] However, the main disadvantages of this type of probe are: 1) the toxicity of copper ions is relatively large; 2) the fluorescence background is large, especially It is the autofluorescence of biological cells and tissues, which is not conducive to large-scale promotion

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
  • Bio-orthogonal activated time-resolved response type rare earth probe and its preparation method and application
  • Bio-orthogonal activated time-resolved response type rare earth probe and its preparation method and application
  • Bio-orthogonal activated time-resolved response type rare earth probe and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] (1) The synthesis steps of the bio-orthogonal activated time-resolved response type rare earth probes provided by the present invention are as follows: figure 1 As shown, the details are as follows: IX 1 Intermediate S 1 Preparation: 7-amino-2-keto-4-quinolinic acid (43.6mg, 0.2mmol) was mixed with N-hydroxysuccinimide NHS (115.0mg, 1.0mmol), 1-(3-dimethyl Aminopropyl)-3-ethylcarbodiimide hydrochloride EDC·HCl (190.8mg, 1.0mmol) was dissolved in 10mL N,N-dimethylformamide and stirred at room temperature overnight for carboxyl activation. Precipitate with saturated saline, centrifuge and dry to obtain a tan solid (i.e. activated 7-amino-2-keto-4-quinolinic acid); continue to mix the tan solid with 4-(6-methyl-1,2, 4,5-tetrazin-3-yl)benzylamine (40.2mg, 0.2mmol) was stirred in N,N-dimethylformamide at room temperature overnight, the solvent was removed, and the red solid S was obtained by silica gel chromatography. 1 (66.8 mg), yield 83.3%.

[0045] characterizing dat...

Embodiment 2

[0049] (1) IX 1 Intermediate S 2 Preparation: at room temperature, remove the red solid S obtained in Example 1 1 (20.5mg, 0.05mmol), diethyltriaminepentaacetic acid DTPA (19.7mg, 0.05mmol) were dissolved in 3mL dimethyl sulfoxide, stirred for overnight reaction, and then precipitated with ether and dried to obtain solid S 2 (34.3 mg), yield 88.4%.

[0050] Characterization data: ICP-MS ( Figure 5 ):calcd.for[M + ]776.76,found:713.40.

[0051] (2) IX 1 Preparation: at room temperature, remove the solid S obtained in step (1) 2 (7.8mg, 0.01mmol), terbium chloride hexahydrate TbCl 3 ·6H 2 O (3.8 mg, 0.01 mmol) was dissolved in 1 mL dimethyl sulfoxide / water (volume ratio, 1:1), stirred overnight for reaction, and then precipitated and dried with ether to obtain rare earth probe IX 1 (8.41 mg), yield 90.1%.

[0052] Characterization data: ICP-MS ( Figure 6 ):calcd.for[M + ]931.66, found: 931.20.

Embodiment 3

[0054] (1) IX 2 Intermediate S 3 , S 4 Preparation: Dissolve the activated 7-amino-2-keto-4-quinolinic acid (33.2 mg, 0.1 mmol) in Example 1 in 10 mL of N,N-diamine containing ethylenediamine (6 mg, 0.1 mmol) In methylformamide, stirred overnight at room temperature, after the completion of the reaction was monitored by TCL, 4-(6-methyl-1,2,4,5-tetrazin-3-yl)benzylamine (30.1 mg, 0.1 mmol) with succinic anhydride (10.0mg, 0.1mmol) was stirred overnight at room temperature in N,N-dimethylformamide, the solvent was removed, and the red solid S was obtained by silica gel chromatography. 3 . At room temperature, the red solid S 3 , diethyltriaminepentaacetic acid (39.5mg, 0.1mmol) was dissolved in 4mL dimethyl sulfoxide, stirred for overnight reaction, then precipitated and dried with ether to obtain its solid S 4 (68.3 mg), yield 74.4%.

[0055] S 3 Characterization data: ICP-MS ( Figure 7 ):calcd.for[M + ]526.59,found:525.10.

[0056] red solid S 3 The structural for...

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 bioorthogonal activated time-resolved responsive rare-earth probe and a preparation method and application thereof, belonging to the technical field of biological markers and imaging. The rare earth probe provided by the present invention contains a tetrazine group, which is combined with Tb 3+ However, the rare earth ion Tb undergoes a bioorthogonal reaction with D-mannose cyclooctene, and the fluorescence resonance energy transfer in the probe cannot occur, resulting in the rare earth ion Tb 3+ The characteristic fluorescence is released, and the probe solution is accompanied by a naked-eye color change from pink to colorless, which can be used for cell membrane imaging of tumor cells; in addition, the rare earth probe has good water solubility and will be used in the solution to tumor cells. The problem of autofluorescence caused by imaging processes such as zebrafish and zebrafish is of great significance.

Description

technical field [0001] The invention belongs to the technical field of biological labeling and imaging, in particular to a bio-orthogonal reaction-based probe with time-resolved fluorescence responsiveness and a preparation method thereof, in particular to a bio-orthogonal activated time-resolved response type rare earth probe and a preparation method thereof and application. Background technique [0002] Bioorthogonal reaction is a type of chemical reaction that can be carried out in living cells without interfering with the biochemical reaction of the organism itself, mainly including azide and alkynyl or phosphoric acid groups, tetrazine and alkenes or trans-cyclooctene , has the characteristics of high selectivity and rapid response. Therefore, the development of bioorthogonal reaction probes is very necessary in broadening biological applications. [0003] At present, this reaction combined with organic fluorescent small molecule probes and its imaging technology has ...

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): C07D403/12C09K11/06G01N21/64A61K49/00
CPCA61K49/0021C07D403/12C09K11/06C09K2211/1007C09K2211/1029C09K2211/1074C09K2211/182G01N21/6428G01N21/6458
Inventor 张涛邢达郑举敦
Owner SOUTH CHINA NORMAL UNIVERSITY
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