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Material for constructing temperature-responsive aggregates in cells and preparation method and application of material

A temperature-responsive and aggregate technology, which is applied in the field of materials for constructing temperature-responsive aggregates in cells and their preparation, and achieves the effect of good application prospects.

Active Publication Date: 2017-09-08
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

However, the precise preparation of large-volume nanomaterials remains a great challenge

Method used

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  • Material for constructing temperature-responsive aggregates in cells and preparation method and application of material
  • Material for constructing temperature-responsive aggregates in cells and preparation method and application of material
  • Material for constructing temperature-responsive aggregates in cells and preparation method and application of material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] In this embodiment, the polypeptide sequence (including responsive polypeptide and membrane-penetrating peptide) in the material for constructing temperature-responsive aggregates in the cells is Cys-Lys-Asp-Glu-Val-Asp-Gly-Arg-Lys- Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Gln, the thermosensitive polymer is poly-N-isopropylacrylamide, the synthesis method is as follows:

[0054] (1) Synthetic responsive polypeptide Cys-Lys-Asp-Glu-Val-Asp-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Gln, namely

[0055]

[0056] The polypeptide was synthesized by Fmoc solid-phase synthesis method: Wang resin with a modification density of 0.35mM was selected for synthesis, in which the C-terminus of the first amino acid (glutamine) was immobilized on the resin, and the N-terminus was protected by Fmoc. The Fmoc protection was deprotected with 20% (v / v) hexahydropyridine in DMF, and the deprotection result was tested by ninhydrin test. Then the carboxyl group of the next amino acid was treate...

Embodiment 2

[0066] The only difference from Example 1 is that in the synthesis process of step (1), Boc-Cys(Trt)-OH is used for the last amino acid, and the amino group of the Lys side chain connected to the Cys is protected by Dde. After removing Dde with 2% hydrazine hydrate, the carboxyl group of the signal molecule Cy5 is connected with the amino group of the side chain of Lys by an amide bond. Then, through the same step (2) and step (3) as in Example 1, the material for constructing temperature-responsive aggregates in cells with signal molecules was prepared.

Embodiment 3

[0068] In this example, the polypeptide sequence CK(S-S)DGRKKRRQRRRPQ with the following structure was synthesized by the same solid-phase synthesis method as step (1) in Example 2:

[0069]

[0070] Figure 6 It is the HPLC spectrogram of the polypeptide sequence CK(S-S)DGRKKRRQRRRPQ, from which it can be seen that the retention time of the polypeptide in the mobile phase is 17.6 minutes.

[0071] Figure 7It is the MALDI-TOF spectrum of the polypeptide sequence CK(S-S)DGRKKRRQRRRPQ, from which it can be seen that the molecular weight of the polypeptide is 2131.5.

[0072] Using the polypeptide sequence CK(S-S)DGRKKRRQRRRPQ, the material for constructing temperature-responsive aggregates in cells was finally prepared through the same steps as step (2) and step (3) in Example 1.

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Abstract

The invention provides a material for constructing temperature-responsive aggregates in cells and a preparation method and application of the material. The material for constructing the temperature-responsive aggregates in the cells comprises a polypeptide sequence composed of responsive polypeptide and cell-penetrating peptide, and a temperature-sensitive polymer, wherein the responsive polypeptide of the polypeptide sequence is connected to the temperature-sensitive polymer. The material for constructing the temperature-responsive aggregates in the cells enters the cells with the assistance of the cell-penetrating peptide and responds to specific stimulating signals in the cells after entering the cells, the hydrophilic polypeptide is removed, the phase transition temperature of the material is changed into the physiological temperature of 37 DEG C or below, collapse and aggregation of the material are caused finally, and the nano-aggregates are formed. With the strategy, long-time retention of the material in the cells can be achieved, and the material has good application prospects in live cell imaging or high-retention-rate drug delivery.

Description

technical field [0001] The invention belongs to the technical field of polymer materials, and relates to a material for constructing temperature-responsive aggregates in cells, a preparation method and application thereof. Background technique [0002] The bioavailability of nanomaterials plays an important role in the application of materials for bioapplications. To improve the biological functions of nanomaterials, various strategies have been developed to enhance the accumulation and retention of nanomaterials at target sites, for example, high permeability and retention (EPR) effect, active targeting mechanism, long-lasting blood circulation, etc. Based on the above concepts, efforts have been made to design and synthesize nanomaterials with easily controllable physicochemical properties, such as size, morphology, charge, surface chemistry, payload, and stability. However, the precise preparation of large-volume nanomaterials remains a great challenge. [0003] In addi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/22A61K49/00A61K47/69A61K47/64C08G81/02A61K31/196A61K31/519
CPCA61K31/196A61K31/519A61K49/0021A61K49/0032A61K49/0073A61K49/226C08G81/02
Inventor 王浩乔圣林马洋李莉莉
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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