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Self-assembly material for in-situ construction of artificial extracellular matrix as well as preparation method and application of self-assembly material

A self-assembly, exogenous matrix technology, applied in the field of biomedicine

Active Publication Date: 2020-08-07
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In summary, there are still relatively few therapeutic strategies for inhibiting tumor metastasis in the prior art. Therefore, it is very important to develop a new type of therapeutic strategy that can inhibit tumor metastasis with significant curative effect

Method used

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  • Self-assembly material for in-situ construction of artificial extracellular matrix as well as preparation method and application of self-assembly material
  • Self-assembly material for in-situ construction of artificial extracellular matrix as well as preparation method and application of self-assembly material
  • Self-assembly material for in-situ construction of artificial extracellular matrix as well as preparation method and application of self-assembly material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] In this example, a self-assembling material for constructing an artificial extracellular matrix in situ is constructed. The self-assembling material is composed of a targeting peptide, a self-assembling polypeptide and a bispyrene fluorescent signal molecule. Its chemical structure is as follows;

[0097]

[0098] Its preparation method comprises the following steps:

[0099](1) Swell Wang resin with N,N-dimethylformamide;

[0100] (2) Using the amino acid whose terminal amino group is protected by Fmoc, the side chain amino group obtained by Boc protection, and the bispyrene fluorescent signal molecule whose terminal amino group is protected by Fmoc as raw materials, first, according to the amino acid sequence of Ala-Lys-Glu-Arg-Cys, the Add alanine to Wang resin, carry out coupling reaction and connection with Wang resin; remove the Fmoc protecting group on alanine, carry out coupling reaction and connection between lysine and alanine; until Ala-Lys- Condensation ...

Embodiment 2

[0108] This embodiment prepares self-assembled nanoparticle solution and nanofiber dispersion, the specific method is as follows:

[0109] The self-assembled material prepared in Example 1 was dissolved in DMSO solvent (the concentration of the self-assembled material was 3×10 -3 M), take the above solution and place it in a centrifuge tube, then slowly add deionized water into the centrifuge tube to prepare mixed solutions with different water contents (0%, 98%), and mix the self-assembly material monomer solution (water content 0%) %) and self-assembled nanoparticle solution (water content 98%), carry out the fluorescence detection with the fluorescence spectrophotometer respectively, and the fluorescence spectrum is as follows figure 2 shown by figure 2 It can be seen that the fluorescence intensity of the self-assembled nanoparticle solution (with a water content of 98%) peaks at 525 nm, indicating the formation of J-type aggregates of bispyrene fluorescent molecules. ...

Embodiment 3

[0114] Laser confocal test:

[0115] In this example, human breast cancer cell MDA-MB-231 and mouse breast cancer cell 4T1 were used as cell models, respectively, in DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin Medium culture, the culture temperature is 37.0 ℃, CO 2 The concentration is 5.0%. When the cells were cultured to the logarithmic growth phase and the cells were in good condition, the dispersed cells were digested with trypsin for 3 min, centrifuged at 1000 r / min for 3 min, and the supernatant was discarded. Each milliliter contains 10 4 The cell suspension of two cells was added to the laser confocal small dish, cultivated for 24h, and then respectively added 1mL of the 30μM self-assembled material prepared in Example 1 to each confocal dish, put into the cell culture incubator and cultivated for 2h, Pour off the supernatant and wash it three times with PBS, add an appropriate amount of PBS, and perform single-photo...

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Abstract

The invention relates to a self-assembly material for in-situ construction of an artificial extracellular matrix, and a preparation method and application thereof. The self-assembly material is composed of a targeting peptide, a self-assembly polypeptide and a dipyrene fluorescence signal molecule, and the chemical structure of the self-assembly material is shown as a formula (I); wherein R1 is from a self-assembled polypeptide with multiple hydrogen bonds in a molecule; r2 and R3 are both from a tumor targeting peptide. The preparation method comprises the step of synthesizing the self-assembled material by taking amino acid and dipyrene fluorescence signal molecules as raw materials through a solid-phase synthesis method. The self-assembly material can actively target a tumor site; meanwhile, the tumor part can be induced to deform; the artificial extracellular matrix is connected with a natural extracellular matrix to jointly form a long-term barrier to inhibit tumor metastasis andinvasion, and the artificial extracellular matrix and the natural extracellular matrix can compete for binding sites of integrin, reduce expression of metal matrix protease and better inhibit tumor metastasis.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a self-assembly material and its preparation method and application, in particular to a self-assembly material for in-situ construction of an artificial extracellular matrix and its preparation method and application. Background technique [0002] Tumor refers to the new organism formed by the proliferation of local tissue cells under the action of various tumorigenic factors. Tumors are divided into benign tumors and malignant tumors. Malignant tumors seriously endanger people's health. The reason for high cancer mortality is mainly due to rapid tumor metastasis. Therefore, inhibiting tumor metastasis is a key factor in tumor treatment. Tumor metastasis is an extremely complex process, among which, the inhibition of tumor metastasis by inhibiting the activity of matrix metalloproteinases is a common strategy, because matrix metalloproteinases degrade the extracellular matrix is ​​the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/64A61K47/69A61K49/00A61P35/00A61P35/04
CPCA61K47/64A61K49/0021A61K47/6953A61P35/00A61P35/04A61K49/0056A61K49/0069Y02P20/55
Inventor 王浩王磊贺平平
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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