Cell penetrating peptide with neutral charge and application of cell penetrating peptide as delivery carrier in cells

A technology that penetrates peptides and cells, applied in the field of biomedicine, and can solve problems such as low efficiency

Inactive Publication Date: 2018-11-06
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In summary, most of the currently known molecular sequences of CPPs contain more basic amino acids (such as arginine and lysine), and their internalization is strongly dependent on the interaction between positively charged basic amino acids and cell membranes under physiological conditions. The interaction of externally negatively charged heparan sulfate, while most of the internalization will be confined to endocytic vesicles, and the intracellular release process is inefficient

Method used

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  • Cell penetrating peptide with neutral charge and application of cell penetrating peptide as delivery carrier in cells
  • Cell penetrating peptide with neutral charge and application of cell penetrating peptide as delivery carrier in cells
  • Cell penetrating peptide with neutral charge and application of cell penetrating peptide as delivery carrier in cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A neutrally charged cell penetrating peptide with a net charge of 0 under physiological conditions. NYNSHNV、WHAPSHH、PAHGTQQ、QSNQHTV、RTSDSHF、HDGRLPW、TTSHNMH、SSTYPHY、GNPSLNQ、HHFSAQH、NHYLSTQ、ANHGVRE、WPNSTST、QDSRFHV、HGVWSQQ、NWYQSLT、PTLHSSQ、EAHKTLS、SSTKDSV、QPTTSYF、TAASHWN、TTTTSNA、AREPCLN、HSGVPHG、HNVSGGS、 HPETLVK, NTPYLTS, DATYGKL, GQGVWPY, ERAGGLL, NSHNVYI, HAPPSSL, DAVRLHW, PILPHGQ, HGAGTAS, APLGTPH, VHNQLQL, PYSSAAS, GPGTSFG, STQSALS, NPGLSSF, STSTAMW, AINGHAS, VPTPSAS, NLGSSYL, WSVPSLYTPVH, TSCMPLST, TL LPSFIHQ, TTYGAGV, LDGVKPV, LTGPAHL, TVLPYWA, WFISSSA, VPWPTLV, LGAGAAT, TAAGITL, NFIAVSA, or GVVPILV.

[0030] Experiments have found that the neutrally charged cell-penetrating peptide can be used as a delivery carrier in mammalian cells. Due to its good cell internalization performance, it can be used as a medical application for the delivery of therapeutic drug carriers in cells, including as a delivery treatment for central degenerative diseases, cerebral ischemia, dr...

Embodiment 2

[0039] In this example, three rounds of whole-cell screening of human cervical adenocarcinoma Hela cells were performed using a phage display peptide library, and 152 cell-penetrating peptide gene sequences were obtained, including 74 neutrally charged penetrating peptides, and neutrally charged peptides were obtained. The amino acid sequence of the penetrating peptide and its internalization performance were verified. The detailed process is as follows:

[0040] 1. Screening of cell-penetrating peptides by phage display technology

[0041] Hela (human cervical adenocarcinoma) cells were cultured in complete medium containing 5% BSA (fetal bovine serum)-DMEM (Dulbecco's modified Eagle's medium) at 37°C, 5% CO 2 Cultivated in an incubator.

[0042] The day before the screening, the cells were seeded in 10cm dishes, and the cells could be used for screening after the cells grew well until the single layer adhered to the bottom of the plate. Discard the culture medium in the 1...

Embodiment 3

[0062] This embodiment provides a homologue of the neutrally charged cell penetrating peptide as in Example 1 or 2, the homologue is any heptapeptide whose amino acid sequence homology is greater than 60%; or the homologue is a neutrally charged peptide containing Any polypeptide or protein of a charged cell penetrating peptide; or a homologue is any polypeptide or protein comprising a homologue of a neutrally charged cell penetrating peptide; or a homologue is a neutrally charged cell penetrating peptide Functional backbone of polypeptides.

[0063] Experiments have found that the homologue of the neutrally charged cell-penetrating peptide can be used as a delivery carrier in mammalian cells. Due to its good cell internalization performance, it can be used as a medical application for the delivery of therapeutic drug carriers in cells, including as a delivery treatment for central degenerative diseases, cerebral ischemia, drug addiction, myocardial infarction, malignant tumor...

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Abstract

The invention provides a cell penetrating peptide with neutral charge, wherein the net charge of the cell penetrating peptide under physiological conditions is 0. The invention also provides a specific amino acid sequence. The invention also provides application of the cell penetrating peptide with neutral charge as a delivery carrier in cells, medical application of the cell penetrating peptide with neutral charge as the delivery carrier of therapeutic drugs in cells, and application of the cell penetrating peptide with neutral charge as a contrast agent in the medical imaging technology of the delivery carrier in cells. Furthermore, homologues of the cell penetrating peptide with neutral charge also have cell penetrating performance and can medically serve as the delivery carrier in cells as well as serve as the delivery carrier of the therapeutic drugs in cells.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a class of neutrally charged cell-penetrating peptides and their application as a delivery carrier in mammalian cells in the fields of biology and medicine. technical background [0002] The cell membrane is a semipermeable barrier between the cell and the extracellular environment, with selective permeability so that the cell maintains a constant internal environment. While this phospholipid bilayer is essential for cell survival and function, it poses challenges for the exchange of cargo molecules inside and outside the cell. Since proteins, polypeptides, nucleotides and other pharmaceutical macromolecules and imaging agents must reach the interior of the cell to play their corresponding roles, it is very necessary to realize the transmembrane transport of these substances. [0003] At present, the technologies that can transport macromolecules into cells mainly include electroporat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K47/42A61K49/22A61K51/08
CPCA61K47/42A61K49/22A61K51/08C07K7/06
Inventor 姜勇刘芸罗海华
Owner SOUTHERN MEDICAL UNIVERSITY
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