CN-121064296-B - Polypeptide with transfection activity and application thereof

Abstract

The invention discloses a polypeptide with transfection activity and application thereof, relating to the technical field of polypeptides, the same as liposomes can encapsulate nucleic acid fragments in vitro and activate membrane receptor transport into the cytoplasm. Compared with liposome, the polypeptide has lower toxicity and is more stable in vivo.

Inventors

• XU SONGHUI
• LI ZHONGQI
• YUAN RUIZE
• HE YIFAN

Assignees

• 南昌大学第一附属医院

Dates

Publication Date

20260505

Application Date

20251104

Claims (2)

1. 1. Use of a polypeptide having transfection activity for the preparation of a transfection preparation, characterized in that the polypeptide has the following structure: ; wherein X is-Arg-His-His-Lys-His-His-Lys-His-His-His.
2. 2. The use according to claim 1, wherein the transfection formulation is an injection or a spread.

Description

Polypeptide with transfection activity and application thereof Technical Field The invention relates to the technical field of polypeptides, in particular to a polypeptide with transfection activity and application thereof. Background Transfection (transfection) is the process by which a cell actively or passively introduces exogenous nucleic acid fragments under certain conditions to obtain a new phenotype. Transfection is widely applied in the fields of scientific research, disease treatment, vaccine coating and the like, and the current transfection reagents mainly comprise liposome reagents such as PEI, LIPO2000 and the like. PEI is an artificially synthesized organic macromolecular cationic polymer, while Lipo2000 and Lipo3000 belong to a cationic liposome transfection reagent, and negatively charged nucleic acid molecules are bound to positively charged liposomes to form nucleic acid-cationic liposome complexes, so that the nucleic acid-cationic liposome complexes are adsorbed to negatively charged cell membrane surfaces and introduced into cells by activating membrane receptor endocytosis. There are numerous reports that liposomes themselves are involved in cellular physiological activities, causing up-or down-regulation of gene expression, such as in PKC (protein kinase C) pathway regulation, such as inhibition of ATPase activity, such as by acting on the mitochondrial membrane, transfection of siRNA, causing off-target effects, and the like. The size of cytotoxicity often means the size of the effects on the physiological activities of the cells, which are the root cause of cytotoxicity. Traditional lipofection agents (e.g., lipo 2000) are easily broken down after entering the living body and accumulate in lung tissue, possibly causing inflammatory reactions or toxicity problems, and thus cannot be directly used for nucleic acid delivery in living animals. Disclosure of Invention The invention aims to at least solve one of the technical problems in the prior art and provides a polypeptide with transfection activity and application thereof. The technical scheme of the invention is as follows: A polypeptide with transfection activity, wherein the amino acid amino residue of the middle main chain is linked with a branched side chain, and the core structure of each branched side chain is-Arg-His-His-Lys-His-His-His-Lys-His-His. Preferably, the amino acid amino residues of the intermediate backbone are linked to a branching side chain, each branching side chain having the core structure-Arg-His-His-Lys-His-His-Lys-His-His-His (abbreviated RHHKHHKHHHKHH as shown in the amino acid sequence table SEQ ID NO. 1), abbreviated X, having the structure: 。 preferably, the preparation method comprises the following steps: Weighing resin, adding DMF to swell for more than half an hour, pumping off DMF, carrying out Fmoc deprotection reaction by using a deprotection reagent, pumping off the deprotection reagent in a shaking table for 10min, washing 3 times by using DMF, then carrying out amino acid condensation reaction, respectively taking corresponding amino acid and condensation reagent, dissolving by using reaction liquid, putting into a reactor, stirring for reaction to obtain peptide resin after amino acid condensation, and repeating the Fmoc deprotection and amino acid condensation reaction steps until the last amino acid reaction is finished to obtain target peptide. Preferably, the Resin is Cl-Resin chloride ion exchange Resin; the deprotection reagent is prepared by dissolving 20wt% piperidine solution in N, N-Dimethylformamide (DMF); the condensation reagent is a mixed solution of N, N-diisopropylcarbodiimide and 1-hydroxybenzotriazole; The invention also discloses application of the polypeptide in preparing a cell transfection preparation. Preferably, the formulation is an injection or a spread. The invention has the beneficial effects that the invention synthesizes a novel peptide, which can wrap the nucleic acid fragment in vitro as liposome and transfer the nucleic acid fragment into cytoplasm, and the polypeptide is a biological macromolecule, and has more stability and lower toxicity than liposome. The invention firstly attracts the surface of a cell membrane through positive and negative charges, then enters cells through combining with a cell surface NPR receptor NRP-1 to have certain biological chemotactic tumor enrichment characteristics, the NRP-1 often presents a high expression state in various malignant tumors, and the high expression is usually closely related to the progress, angiogenesis, metastasis and bad prognosis of the tumors. Drawings FIG. 1 shows the effect of electrophoresis detection of plasmids got, siAR coated with the polypeptide xsh 007; FIG. 2 shows the effect of in vitro transfection of 2mg of plasmid per six well plate single well over-expression of molecules of interest; FIG. 3 is an in vitro transfection siRNF41 knockdown effect; FIG. 4 is an in vitro transfection siAR k