Protein nanoparticles carrying antitumor peptide and preparation method and application of protein nanoparticles

Abstract

The invention belongs to the technical field of biomedicines and in particular relates to protein nanoparticles carrying an antitumor peptide and a preparation method and application of the protein nanoparticles. The preparation method comprises the following steps: according to gene sequences of a pET25b(+)-HSP16.5 expression vector, synthesizing a single stranded DNA (deoxyribonucleic acid) sequence corresponding to an apoptosis-promoting peptide, namely KLAK peptide, performing annealing treatment, performing HindIII and XhoI double-digestion on the product and the vector, performing gel recycling on the digestion product, connecting the KLAK sequence with the vector, transforming the connection product into a competence of DH5a escherichia coli, picking a monoclonal bacterial colony, performing overnight bacterium shaking, extracting a plasmid, and performing sequencing; and transforming a pET25b(+)-HSP-KLAK plasmid with correct sequencing into a competence of BL21 escherichia coli, amplifying culture, performing ultrasonic bacterium crushing so as to obtain a corresponding crude product, performing purification, and performing dialysis, concentration and freeze-drying, so as to obtain an HSP-KLAK protein. The heat shock protein nanoparticles carrying the antitumor peptide, which are provided by the invention, can be rapidly taken in by tumor cells, after being released inlysosome, a targeting mitochondrion plays a role of tumor killing, the protein of the medicine carrier self can be rapidly degraded by cells, and thus a good tumor treatment effect can be achieved.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a protein nanoparticle carrying an antitumor 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 multiple stages of development. The serious toxicity and side effects of traditional chemotherapy drugs during clinical use and the multi-drug resistance of tumor cells are seriously limited. Their application in tumor therapy. [0003] Polypeptide is a small "protein" composed of a small amount of amino acids (10-100), which has the advantages of small molecular weight, easy modification, clear structure, clear mechanism of action, and activity close to that of protein. It combines the advantages of chemical drugs and protein drugs. Therefore, It has become one of the hot spots in the current research and developmen...

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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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