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Cell-penetrating peptide and pharmaceutical composition and preparation method and application thereof

A technology of membrane-penetrating peptides and compositions, which is applied in the field of biomedicine and can solve problems such as reduced membrane-penetrating efficiency and inability to transport substances in secondary structures

Active Publication Date: 2013-11-06
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the shortcomings of the existing membrane-penetrating peptides, which are prone to secondary structure when the peptide segment is too long; or when the peptide segment itself is too short, its membrane-penetrating efficiency is significantly reduced, and provides A membrane-penetrating peptide with short length and high transport efficiency and its application, as well as a pharmaceutical composition and its preparation method

Method used

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  • Cell-penetrating peptide and pharmaceutical composition and preparation method and application thereof
  • Cell-penetrating peptide and pharmaceutical composition and preparation method and application thereof
  • Cell-penetrating peptide and pharmaceutical composition and preparation method and application thereof

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preparation example Construction

[0030] In the fourth aspect, according to a preferred embodiment of the present invention, a method for preparing a pharmaceutical composition is provided, wherein the method includes: mixing the penetrating peptide provided by the present invention and siRNA in a buffer with a pH value of 7.0-8.5 contact in liquid.

[0031] Preferably, the molar ratio of the penetrating peptide to siRNA is 20-100:1.

[0032] According to the present invention, the total amount of the penetrating peptide and siRNA can be adaptively adjusted according to the efficiency of the reaction and the actual requirement of the pharmaceutical composition. Preferably, the pH value per milliliter is 7.0-8.5 buffer solution, the total amount of the penetrating peptide and siRNA added is 30-70 μmol.

[0033] According to the present invention, the buffer is various buffers that can be used in pharmaceutical compositions well known to those skilled in the art, for example, Tris-hydrochloric acid buffer, citr...

Embodiment 1-5

[0040] This example is used to illustrate the penetrating peptide provided by the present invention

[0041]Entrust Beijing Hengyu Vision Biotechnology Co., Ltd. to synthesize SEQ ID No: 1 (RRWQWR), SEQ ID No: 2 (RRWQWRMKKL), SEQ ID No: 3 (FKCRRWQWRMKKL), SEQ ID No: 4 (RRWQWRMKKLGAPSITCVRR) and SEQ ID No : 5 (FKCRRWQWRMKKLGAPSITCVRRAF), the amino acid sequences are denoted as bLfcin1, bLfcin2, bLfcin3, bLfcin4 and bLfcin5, respectively.

[0042] A small amount of the above-mentioned five membrane-penetrating peptides were identified by mass spectrometry, and the molecular weight obtained by mass spectrometry was consistent with the calculated value obtained according to the sequence, which proved that the five membrane-penetrating peptides were the membrane-penetrating peptides provided by the present invention.

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Abstract

The invention discloses a cell-penetrating peptide and its application in the preparation of a medicine for entering cells, wherein the cell-penetrating peptide is selected from a group consisting of amino acid sequences as shown in SEQ ID No:1, SEQ ID No:2, SEQ ID No:3, SEQ ID No:4 and SEQ ID No:5. The invention also discloses a pharmaceutical composition and a preparation method of the pharmaceutical composition, wherein the pharmaceutical composition contains the cell-penetrating peptide provided by the invention and siRNA; and the preparation method comprises a step of letting the cell-penetrating peptide provided by the invention and siRNA contact in a buffer at pH of 7.0-8.5. The invention also provides the pharmaceutical composition prepared by the preparation method.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular to a membrane-penetrating peptide, an application of the membrane-penetrating peptide, a pharmaceutical composition, a preparation method of the pharmaceutical composition, and an application of the pharmaceutical composition. Background technique [0002] Cell-penetrating peptides (CPPs), usually composed of 5-30 amino acids, can mediate macromolecular substances through the cell membrane into the cytoplasm and nucleus. CPP has attracted extensive attention from scholars at home and abroad because of its low dosage, short transport time, and no toxic side effects in the process of mediating macromolecular substances through the cell membrane into the cytoplasm and nucleus. In recent years, CPPs have been widely used to transport polypeptides, proteins, oligonucleotides, DNA plasmids, liposomes and metal ions in vivo and in vitro, and many of them have even been applied to the cli...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K7/08C07K14/00A61K47/42A61K48/00
Inventor 方冰张明郭慧媛姜鹭任发政
Owner CHINA AGRI UNIV
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