Molecular probe, and preparation method and application thereof

A molecular probe and molecular technology, which are applied in the field of molecular probes and their preparation to achieve the effects of good specificity and signal stability, enrichment of autophagy detection methods, and broad application prospects

Active Publication Date: 2017-01-04
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, molecular probes with specific responses for in vivo cell autophagy imaging are rarely reported. Therefore, based on the AIE characteristics of fluorescent molecules, molecular probes with high biosafety performance, sensitivity and specificity are developed to be used as in vivo and in vitro cell autophagy. The diagnostic imaging agent of autophagy will provide the possibility for the diagnosis and treatment of autophagy-related diseases

Method used

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  • Molecular probe, and preparation method and application thereof
  • Molecular probe, and preparation method and application thereof
  • Molecular probe, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1: the preparation of molecular probe

[0054] 1. Taking the bipyrene molecular probe with AIE effect as an example, the responsive polypeptide sequence is:

[0055] Gly-Lys-Gly-Ser-Phe-Gly-Phe-Thr-Gly.

[0056] Polypeptide synthesis was synthesized by Fmoc solid-phase synthesis: (1) Wang resin with a modification density of 0.35mM was used for synthesis, wherein the C-terminus of the first amino acid (glycine) was fixed on the resin, and the N-terminus was protected by Fmoc. / v) The DMF solution of hexahydropyridine removes the Fmoc protection, and then uses the ninhydrin test method to test the deprotection result;

[0057] (2) Add the carboxyl group of the next amino acid with a DMF solution of 0.4M 4-methylmorpholine and 0.4M benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate Activated and added to the deprotected resin to react for 2 hours, wherein the second amino acid adopts the side chain amino group as Dde-protected lysine, after comple...

Embodiment 2

[0072] Example 2: In vivo photoacoustic imaging experiment of autophagy

[0073] Taking the imaging of autophagy detection in mouse tumor models by the molecular probe of purpurin 18, which has the characteristics of aggregation and enhanced photoacoustic signal, as an example. The molecular probe chemical structure is as follows:

[0074]

[0075] Select BALB / c nude mice of 6-7 weeks, and inject 1×10 7 U87 glioma cells were used to establish a mouse hind leg tumor model. Inject 1 μM autophagy inducer rapamycin 100 μL through the tail vein to activate the tumor model, and then inject 200 μL of the molecular probe with a concentration of 100 μg / mL through the tail vein, and select 0.25, 0.5, 1, 2, 4, 6 , 7, 9, 12 and 24 hour time points, observe the imaging of the autophagy level of the tumor site at different time points.

[0076] The result is as Figure 9 As shown, no fluorescent signal was detected at any time period in the tumor site of mice treated with normal sali...

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Abstract

The invention provides a molecular probe, and a preparation method and application thereof; the molecular probe comprises water-soluble carrier molecules, responsive polypeptides and signal molecules, wherein the responsive polypeptides have a sequence of (Xaa)m(Thr-Phe-Gly-Phe)(Xaa)nLys(Xaa) i, and wherein Xaa refers to any amino acid, and i, n and m are independently 0-4; the water-soluble carrier molecules and the signal molecules are connected with the responsive polypeptides through amido bonds. The molecular probe herein has good specificity and signal stability, provides richer autophagy detection means and has a promising application prospect.

Description

technical field [0001] The invention belongs to the technical field of molecular probes, and relates to a molecular probe and its preparation method and application. Background technique [0002] Autophagy is a process in which cells rely on lysosomes to transport damaged, denatured or aging proteins and organelles to lysosomes for digestion and degradation. Under normal physiological conditions, autophagy helps cells maintain homeostasis; when stress occurs, autophagy prevents the accumulation of toxic or carcinogenic damaged proteins and organelles, inhibits cell canceration, and maintains the stability of the intracellular environment and metabolic balance. Cell autophagy is extremely critical in the growth and development of organisms, cell differentiation, and responses to environmental stress. It participates in processes such as cell survival, death, pathogen clearance, and antigen presentation, and is associated with cancer, infection, and neurodegenerative diseases....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K49/22G01N21/64
CPCA61K49/0021A61K49/0036A61K49/0054A61K49/0056A61K49/221G01N21/6428G01N21/6486G01N2021/6432
Inventor 王浩林耀新王磊
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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