Cell-penetrating peptide and pharmaceutical composition and preparation method and application thereof
A technology for penetrating peptides and compositions, which is applied in the field of biomedicine and can solve the problems of inability to transport substances in secondary structures and reducing the efficiency of penetrating membranes.
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[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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