A kind of protein nanoparticle carrying antitumor peptide and its preparation method and application

Abstract

The invention belongs to the technical field of biomedicine, and relates to a protein nanoparticle carrying an anti-tumor peptide and its preparation method and application; the preparation method is: according to the gene sequence of the pET25b(+)-HSP16.5 expression vector, synthesizing the pro-apoptotic peptide The single-stranded DNA sequence corresponding to the KLAK peptide, after the annealing treatment, the product and the carrier were subjected to HindⅢ and XhoI double enzyme digestion, the digested product was recovered from the gel, the KLAK sequence was connected to the carrier, and the ligated product was transformed into DH5a Escherichia coli competent, picked Shake the monoclonal colony overnight, extract the plasmid for sequencing; transform the correctly sequenced pET25b(+)-HSP-KLAK plasmid into BL21 Escherichia coli competent, expand the culture, sonicate the bacteria to obtain the corresponding crude product, then purify, dialyze and concentrate HSP-KLAK protein is obtained after freeze-drying; the heat shock protein nanoparticles carrying anti-tumor peptides of the present invention can be rapidly taken up by tumor cells, and after being released in lysosomes, target mitochondria to exert tumor-killing functions, while the protein of the drug carrier itself It can be rapidly degraded by cells to achieve a good therapeutic effect on tumors.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a protein nanoparticle carrying an anti-tumor peptide and a preparation method and application thereof. Background technique [0002] Cancer, also known as malignant tumor, is a frequently-occurring disease caused by multiple factors, multiple gene mutations, and multi-stage development. The severe toxic side effects and multi-drug resistance of tumor cells during the clinical use of traditional chemotherapy drugs have severely limited Their use in tumor therapy. [0003] Peptides are small "proteins" composed of a small number of amino acids (10-100), which have the advantages of small molecular weight, easy modification, clear structure, clear mechanism of action, and activity close to that of proteins. They combine the advantages of chemical drugs and protein drugs, so It has become one of the hot spots in the research and development of anticancer drugs. [0004] Antitu...

AI Technical Summary

Problems solved by technology

However, due to its hydrophilicity, it cannot be effectively internalized into tumor cells, so it must be coupled with a receptor or ligand in the cell-penetrating part, and enter the target cell through a receptor-independent and dependent mechanism in order to exert its potential. pro-apoptotic properties

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