Unlock instant, AI-driven research and patent intelligence for your innovation.

A method for detecting mmp9 protein in solution

A technology of MMP9 and solution, which is applied in the field of chemical medicine and can solve problems that have not been reported

Active Publication Date: 2016-07-06
LANZHOU UNIVERSITY
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In recent years, due to the advantages of molecular docking technology in the specific recognition of macromolecules, it has been widely used in the design of drug targets and achieved good results; There is no report on the detection of specific 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 method for detecting mmp9 protein in solution
  • A method for detecting mmp9 protein in solution
  • A method for detecting mmp9 protein in solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1 A method for detecting MMP9 protein in solution, comprising the following steps:

[0069] ⑴ Molecular docking technology was used to design the specific small molecule ligand of MMP9 (water-soluble white solid with a molecular weight of 355 and stable chemical properties). The chemical structural formula of the specific small molecule ligand of MMP9 is as follows (see reference for the design principle Literature XiL, DuJ, LiS, et al.A combined molecular modeling study on gelatinases and their potentin inhibitors [J]. JComputChem.2010,31(1):24-42.):

[0070] .

[0071] (2) Chemical synthesis of small molecule ligands:

[0072] ① Preparation of potassium fluoride-calcium fluoride (KF-CaF 2 ) melt:

[0073] Mix potassium fluoride dihydrate and calcium fluoride at a material ratio of 1:1.5, stir evenly and grind for 1 hour to obtain a powder mixture, which is vacuum-dried at 140°C for 8 hours to obtain potassium fluoride - Calcium Fluoride (KF-CaF 2 ) m...

Embodiment 2

[0113] Embodiment 2 A method for detecting MMP9 protein in solution, comprising the following steps:

[0114] (1) The design of the specific small molecule ligand of MMP9 using molecular docking technology is the same as that in Example 1.

[0115] (2) Chemical synthesis of small molecule ligands:

[0116] ① Preparation of potassium fluoride-calcium fluoride (KF-CaF 2 ) melt:

[0117] Mix potassium fluoride dihydrate and calcium fluoride at a material ratio of 1:2.5, stir evenly and grind for 2 hours to obtain a powder mixture, which is vacuum-dried at 160°C for 5 hours to obtain potassium fluoride - Calcium Fluoride (KF-CaF 2 ) melt.

[0118] ② Add p-toluenesulfonyl chloride, potassium fluoride-calcium fluoride melt in sequence in dry acetonitrile, stir at room temperature for 12 hours, and after the reaction is completed, cover 3 cm thick diatomaceous earth with a sand core funnel to filter to obtain the filtrate. The filtrate was distilled under reduced pressure at 40°C ...

Embodiment 3

[0152] Embodiment 3 A method for detecting MMP9 protein in solution, comprising the following steps:

[0153] (1) The design of the specific small molecule ligand of MMP9 using molecular docking technology is the same as that in Example 1.

[0154] (2) Chemical synthesis of small molecule ligands:

[0155] ① Preparation of potassium fluoride-calcium fluoride (KF-CaF 2 ) melt:

[0156] Mix potassium fluoride dihydrate and calcium fluoride at a ratio of 1:2, stir evenly and grind for 1.5 hours to obtain a powder mixture, which is vacuum-dried at 150°C for 6 hours to obtain fluoride Potassium-calcium fluoride (KF-CaF 2 ) melt.

[0157] ② Add p-toluenesulfonyl chloride, potassium fluoride-calcium fluoride melt in sequence in dry acetonitrile, stir at room temperature for 10 hours, after the reaction is completed, cover the 2.5cm thick diatomaceous earth with a sand core funnel and filter to obtain the filtrate, The filtrate was distilled under reduced pressure at 35°C to obta...

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

PropertyMeasurementUnit
thicknessaaaaaaaaaa
thicknessaaaaaaaaaa
Login to View More

Abstract

The invention relates to a method for detecting MMP9 protein in solution. The method comprises the following steps: (1) designing an MMP9 specific micromolecule ligand; (2) chemically synthesizing the micromolecule ligand, comprising a, preparing KF-CaF2 melt; b, preparing p-toluenesulfonyl fluoride, c, reacting by using catalyst LDA to obtain a compound I; d, removing Boc group on a reactant by HCL gas to obtain a compound II; e, reacting by using catalyst triethylamine to obtain a compound III; f, reacting the compound II with the compound III to obtain a compound IV; g, reacting the compound IV with NaoH solution to obtain a compound V; h, reacting by using catalysts O-(Tetrahydro-2H-pyran-2-yl) hydramine, NMM, HOBT and EDC to obtain a compound VI; and I, removing a protective group from the compound VI to obtain an MMP9 micromolecule ligand; (3) detecting MMP9 in the solution by an SPR method: i, modifying a naked gold film chip; ii, fixing the micromolecule ligand; iii, removing non-specificity micromolecule adsorption; iv, detecting the MMP9 protein in the solution; and v, removing the MMP9 non-specificity protein adsorption. The method is high in sensitivity.

Description

technical field [0001] The invention relates to the field of chemical medicine, in particular to a method for detecting MMP9 protein in a solution. Background technique [0002] Molecular docking technology (Molecular docking technology) simulates the interaction between the specific active center of the biological macromolecule and the small molecule of the ligand at the molecular level, and calculates the binding free energy between the macromolecule and the small molecule, which is used to design new energy. Small molecules that specifically bind to macromolecular substances and have strong affinity can save a lot of time and shorten the experimental process. Docking technology is a method and method commonly used in computer-aided drug design. In recent years, great success has been achieved in drug design. The introduction of molecular docking technology has become a possibility to solve the specific recognition of specific proteins. [0003] Synthetic chemistry is the...

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): G01N21/552
Inventor 王志平刘映前姚小军王静张丽秀祁萍
Owner LANZHOU UNIVERSITY