CAGW polypeptide and TAT-NLS polypeptide-modified gene-loaded composite, preparation method and application thereof

A technology of TAT-NLS and polypeptide modification, which is applied in the field of biomedicine to achieve the effects of enhanced transfection efficiency, improved biocompatibility, and improved endosome escape ability

Active Publication Date: 2019-05-03
QINGHAI UNIV FOR NATITIES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the above-mentioned problems in the prior art, the present invention provides a gene-carrying compound modified by CAGW polypeptide and TAT-NLS polypeptide and its preparation method and application, which can effectively improve the difficulty of the existing gene-carrying compound entering cells and the transgenic complex. The problem of low dyeing efficiency

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  • CAGW polypeptide and TAT-NLS polypeptide-modified gene-loaded composite, preparation method and application thereof
  • CAGW polypeptide and TAT-NLS polypeptide-modified gene-loaded composite, preparation method and application thereof
  • CAGW polypeptide and TAT-NLS polypeptide-modified gene-loaded composite, preparation method and application thereof

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Embodiment 1

[0036] A gene-carrying complex modified by a CAGW polypeptide and a TAT-NLS polypeptide, the preparation method of which comprises the following steps:

[0037] 1. Synthesis of polyethylene glycol-polyethylene lactide copolymer mPEG-b-PLGA (named mP) (b is the abbreviation of block, representing block polymerization)

[0038] Using mPEG as a macromolecular initiator, stannous octoate as a catalyst, initiates the ring-opening copolymerization of DL-LA and GA, the specific process is: weigh 1.956g (0.3912mmol) mPEG, 0.8078g (6.964mmol) glycolide (GA) , 3.209g (22.28mmol) DL-type lactide (DL-LA), put into a dry polymerization tube, add Sn(Oct) 2 Toluene solution (0.4mL, 0.25mol / L), sealed, evacuated, nitrogen, repeated several times, then put into an oil bath, reacted at 110°C for 24h. The obtained product was dissolved in chloroform, precipitated in n-hexane, and the product was purified several times, dried in vacuum at 25°C, and set aside.

[0039] 2. Synthesis of polylactic...

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Abstract

The invention relates to a CAGW polypeptide and TAT-NLS polypeptide-modified gene-loaded composite, a preparation method and an application thereof. The preparation method comprises the following steps: preparing polyethylene glycol-PGLA copolymer mPEG-b-PLGA; preparing a PGLA-g-polyethyleneimine copolymer PLGA-g-PEI; using PLGA-g-PEI as a raw material for preparation of a PLGA-g-PEI-g-CAGW copolymer; performing self-assembly preparing on the targeted copolymer micelles by of PLGA-g-PEI-g-CAGW and mPEG-b-PLGA; adding pDNA and TAT-NLS to the targeted copolymer micelles to prepare the gene-loaded composite. The gene-loaded composite prepared by the invention has specific selectivity to cells, has good penetrating ability and nuclear importing ability, and has high transfection efficiency.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a gene-carrying compound modified by a CAGW polypeptide and a TAT-NLS polypeptide, and a preparation method and application thereof. Background technique [0002] Cardiovascular diseases occur due to endothelial dysfunction or damage, so reconstruction or repair of vascular endothelium is an effective strategy for the treatment of cardiovascular diseases. In many studies, ECs cells were cultured and planted on blood vessels with endothelial damage, so as to achieve the purpose of improving and repairing endothelial function. Wilson et al. used recombinant retrovirus to introduce LacZ gene into canine endothelial cells. After the cells were transfected with the LacZ gene, they were planted on the artificial blood vessel and implanted into the canine great carotid artery. Five weeks later, the results of scanning electron microscopy confirmed that the endothelial c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P9/00
Inventor 朵兴红冯亚凯白凌闯
Owner QINGHAI UNIV FOR NATITIES
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