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High-specificity MMP-14 substrate peptide, preparation method and applications thereof

A MMP-14, high-specificity technology, applied in the field of biological enzyme detection, achieves the effect of simple structure and high safety in vivo

Active Publication Date: 2017-12-22
泊诺(天津)创新医药研究有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, due to the obvious substrate cross-activity between MMPs family proteases, how to improve the specificity of substrate peptides has become a key issue in the development of fluorescent detection probes.

Method used

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  • High-specificity MMP-14 substrate peptide, preparation method and applications thereof
  • High-specificity MMP-14 substrate peptide, preparation method and applications thereof
  • High-specificity MMP-14 substrate peptide, preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-3

[0053] Synthesis sequence: from C-terminal to N-terminal. The synthesis steps are as follows:

[0054] (1) Resin swelling

[0055] Put 2-Chlorotrityl Chloride Resin into the reaction tube, add DCM (15mL / g), and shake for 30min.

[0056] (2) Take the first amino acid

[0057] Filter out the solvent through a sand core, add a 3-fold molar excess of Fmoc-Leu-OH amino acid, add DMF to dissolve, then add a 10-fold molar excess of DIEA, and shake for 60 minutes. Block with methanol.

[0058] (3) Deprotection

[0059] Remove DMF, add 20% piperidine DMF solution (15mL / g) for 5min, remove and add 20% piperidine DMF solution (15mL / g) for 15min.

[0060] (4) Detection

[0061] Take out the piperidine solution, take more than a dozen resins, wash them three times with ethanol, add detection reagents for detection, heat at 105°C-110°C for 5min, and turn dark blue as a positive reaction.

[0062] (5) Washing: DMF (10 mL / g) twice, DCM (10 mL / g) twice, DMF (10 mL / g) twice.

[0063] (6...

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Abstract

The invention belongs to the field of biological enzyme detection, and particularly relates to a high-specificity MMP-14 substrate peptide, a preparation method and applications thereof, wherein the substrate peptide has a structure represented by a formula I, n is an integer of 4-30, and F1 and F2 are respectively a fluorophore and a corresponding quenching group thereof. According to the present invention, the substrate has high specificity, uses the polypeptide and the PEG structure as the basis, and has broad prospects in matrix metalloproteinase detection and cancer detection and treatment. The formula I is defined in the specification.

Description

technical field [0001] The invention relates to the field of biological enzyme detection, in particular to a highly specific MMP-14 substrate peptide and its preparation method and application. Background technique [0002] Matrix metalloproteases (MMPs) are a class of proteolytic enzymes that are highly conserved in natural evolution and whose activity depends on zinc ions and calcium ions. They are widely distributed in plants, vertebrates, and invertebrates. At present, 28 members of this family have been isolated and identified, numbered MMP1-28. Members of this family have a similar structure and are generally composed of five structural domains with different functions, including a hydrophobic signal peptide sequence, a propeptide region (to maintain the stability of the zymogen), a catalytically active region (Zn 2+ binding site), a proline-rich hinge region, and a carboxy-terminal region (relevant to the substrate specificity of the enzyme). The members of MMPs hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K5/117C07K1/04C07K1/06C07K1/16C07K1/107C12Q1/37
CPCC07K5/1024C07K7/08C07K19/00C12Q1/37
Inventor 孙璐杨志民贺慧宁解树平
Owner 泊诺(天津)创新医药研究有限公司
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