Flavone glycoside functionalized magnetic nanometer affinity probe, preparation method and applications thereof, and intracellular target protein capture method

A flavone glycoside, magnetic nanotechnology, applied in nanotechnology, nanomedicine, nanotechnology, etc., can solve the problems of weak interaction, loss of heavy target proteins, etc., to achieve the effect of simplified operation and high protein capture specificity

Active Publication Date: 2018-10-23
INST OF CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are currently two main challenges: on the one hand, the interaction between pharmacologically active molecules and proteins is relatively weak, and conventional methods are difficult to keep the interaction between them from being destroyed; on the other hand, the expression of drug target proteins Many at the trace level, ordinary ESI-MS or MALDI-MS loses the information of many heavy target proteins due to the detection limit problem

Method used

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  • Flavone glycoside functionalized magnetic nanometer affinity probe, preparation method and applications thereof, and intracellular target protein capture method
  • Flavone glycoside functionalized magnetic nanometer affinity probe, preparation method and applications thereof, and intracellular target protein capture method
  • Flavone glycoside functionalized magnetic nanometer affinity probe, preparation method and applications thereof, and intracellular target protein capture method

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preparation example Construction

[0047] The present invention also provides a method for preparing the above-mentioned flavone glycoside functionalized magnetic nano-affinity probe, the method comprising the following steps,

[0048] 1) In the presence of an organic solvent, the flavone glycoside compound is first contacted with the compound of the structure shown in formula (2), to obtain an intermediate compound in which one end of the compound of the structure shown in formula (2) is bonded to the flavone glycoside compound;

[0049] 2) performing a second contact with the intermediate compound obtained in step 1) and the magnetic nanoparticles to obtain a flavone glycoside functionalized magnetic nano-affinity probe whose other end of the compound shown in the formula (2) is connected to the magnetic nanoparticles;

[0050]

[0051] In formula (2), R 1 Represents azido, alkynyl or amino, R 2 Represents an amino group or a BOC-protected amino group, and n is an integer of 2-10.

[0052] Preferably, in...

Embodiment 1

[0086] This Example 1 is used to illustrate the preparation and characterization of the baicalin derivative Qz-1 of the present invention.

[0087] Weigh baicalin (446.2mg, 1mmol, Baicalin, its structure is shown in the above formula (1), the same below), HOBt (175.6mg, 1.3mmol) is placed in a round bottom flask, add 6ml of anhydrous DMF, stir for 30min . Take N in the fridge 3 -PEG 3 -NH 2 (218.3 mg, 1 mmol (1amino-11 azide-3,6,9-trioxa-undecane) was dissolved in 2 ml of DMF, mixed well, added dropwise to the flask, EDCI (249.2 mg, 1.3 mmol) was dissolved in 2 ml of Add water DMF dropwise, add excess TEA (303.3mg, 3mmol) for a period of time, stir and react at room temperature for 72h, add water three times to stop the reaction, CHCl 3 Extracted, dried over night with anhydrous magnesium sulfate, separated and purified by silica gel column chromatography, and dried in an oven to obtain a light yellow powder (baicalin derivative Qz-1 (hereinafter also referred to as Qz-1),...

Embodiment 2

[0092] This Example 2 is used to illustrate the preparation and characterization of the baicalin derivative-ferric iron tetroxide magnetic nano-affinity probe of the present invention (that is, the flavone glycoside functionalized magnetic nano-affinity probe of the present invention).

[0093] The concentration of the alkyne-functionalized magnetic nanoparticles is 30 mg / ml, and the preservation solution is 50% ethanol aqueous solution, which is dispersed uniformly by ultrasonic before use. The baicalin derivative Qz-1 was made up to 5 mM in DMSO and diluted with PBS to contain 1.25% by weight of DMSO before use. Anhydrous copper sulfate was prepared to 5mM with water, sodium vitamin C was prepared to 50mM with three times of water before use. The construction method of baicalin derivative-ferric oxide affinity nanoprobe (MNPs@Qz-1) is as follows: take a clean 1.5ml centrifuge tube, add 100μL (3mg) magnetic nanoparticles into the tube, magnetically separate and discard the pr...

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Abstract

The invention relates to the field of drug target protein enrichment, separation and detection, and discloses a flavone glycoside functionalized magnetic nanometer affinity probe, and further providesa preparation of the flavone glycoside functionalized magnetic nanometer affinity probe, applications of the flavone glycoside functionalized magnetic nanometer affinity probe in the drug target protein capture and/or identification of cells, and a method for capturing an intracellular target protein through the flavone glycoside functionalized magnetic nanometer affinity probe, wherein the flavone glycoside functionalized magnetic nanometer affinity probe comprises magnetic nanoparticles, a coupling structure and a flavone glycoside compound supported on the surface of the magnetic nanoparticle, one end of the coupling structure is coupled to the surface of the magnetic nanoparticles, and the other end of the coupling structure is coupled to the flavone glycoside compound. With the flavone glycoside functionalized magnetic nanometer affinity probe of the present invention, the high-specificity capture can be achieved only with a small amount of the whole protein extraction liquid.

Description

technical field [0001] The present invention relates to the field of drug target protein enrichment, separation and detection, in particular to a flavone glycoside functionalized magnetic nano-affinity probe and its preparation method and its application in the capture and / or identification of drug target protein in cells ; and a method for capturing intracellular target proteins by flavone glycoside functionalized magnetic nano-affinity probes. Background technique [0002] Flavonoids are a class of polyphenols that widely exist in plants, and most of them come from the secondary metabolism of plants. Its basic core is 2-phenyl chromone, which is based on C 6 -C 3 -C 6 way to connect. According to the connection position and chemical properties, there are mainly flavonoids, flavonols, dihydroflavones, anthocyanins, dihydroflavonols, isoflavones, and ercones. A large number of studies have shown that flavonoids have various functions such as anti-free radicals, analgesi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68B82Y30/00B82Y5/00
Inventor 汪福意梁祖青罗群
Owner INST OF CHEM CHINESE ACAD OF SCI
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