Application of cell penetrating peptide Tat-mediated growth factor in transdermal transfer

A growth factor-mediated technology, applied in the field of transdermal drug delivery systems and cosmetics, can solve the problems of difficult-to-explain macromolecular substances, and achieve the effect of increasing the transdermal rate and quantity

Active Publication Date: 2010-05-26
BIOPHARM RES & DEV CENT JINAN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the reverse micelle mechanism is difficult to explain macromolecular substances, such as proteins or substances with larger structures

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Expression and purification of HIV-1-containing transactivator protein Tat protein transduction domain (Tat-protein transductiondomain, Tat-PTD 49-57 ) (Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg) and human epidermal growth factor fusion protein Tat-EGF. The gene fragment encoding Tat-EGF fusion protein was obtained by PCR method, and a 6×His tag was added at the 3' end. After the amplified product was treated with restriction endonuclease BamH I and Nde I, it was connected to the Escherichia coli expression plasmid pET-3c (Novagen, co. , co.Ltd., USA), ampicillin (final concentration 100ug / ml) screened positive clones. The correct recombinant plasmid identified by sequencing was extracted and transformed into Escherichia coli BL21(DE3) (Novagen, co. Ltd., USA), and the transformants were selected by ampicillin resistance (final concentration 100ug / ml). At 37°C, after being induced by IPTG (final concentration: 1mmol / L) for 4 hours, the fusion protein was harvested and separ...

Embodiment 2

[0023] Dissolve 0.1 g of hyaluronic acid in about 80 ml of double-distilled water, then add sodium heparin (1 / 4 of the amount of Tat-EGF fusion protein), 4000.2 g of PEG, stir well and then sterilize under high pressure (0.1Mpa, 121°C for 15 minutes ), cooled to room temperature. The Tat-EGF fusion protein obtained in Example 1 with a purity of more than 98% was sterilized by filtration through a 0.22 μm microporous membrane, and the solution was added to the above mixture, the pH was adjusted to 6.8, and sterilized double-distilled water was added to 100 ml.

Embodiment 3

[0025] Take 0.1g of water-soluble chitosan and dissolve it in an appropriate amount of glacial acetic acid (about 1ml), add about 50ml of double-distilled water, adjust the pH to 6, then add 0.1g of EDTA-2Na, 0.2g of PEG 400, dissolve and then add paraben 0.01g each of the ester and ethylparaben (dissolved by heating with an appropriate amount of water first), stir well, then autoclave (0.1Mpa, 121°C for 15 minutes), and cool to room temperature. Add the Tat-EGF fusion protein with a purity of more than 98% obtained in Example 1, and sterilize it through a 0.22 μm microporous membrane filter into the above mixture, adjust the pH to 6.8, and add sterilized double-distilled water to 100 ml. Dispensed in a spray bottle.

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Abstract

The invention discloses an application of a growth factor series mediated by an active factor Tat protein transcribed by an immunodeficiency virus (HIV-1) in the transdermal transfer, in particular to an application in the fields of transdermal administration and cosmetics. The application is achieved by the following steps of: constructing a fusion protein recombinant expression vector of Tat and a series of active proteins in the growth factor family by the general molecule clone method, purifying with different chromatographic columns to obtain a fusion protein, or linking the Tat to a natural or synthetic phosphatide by the chemical and (or) physical method to prepare a liposome coating the growth factor, forming a transdermal transfer system for the Tat-mediated liposome, matching with proper pharmaceutical or cosmetic auxiliary materials to prepare finished products used for a transdermal administration system or a cosmetics system. The invention can effectively transfer the active proteins of the growth factor with macromolecules through the skin by the Tat mediation and fully exert the effect. The invention provides a novel platform to the application of Tat.

Description

Technical field: [0001] The invention relates to a transdermal delivery system mediated by the membrane-penetrating peptide Tat, which is suitable for the fields of transdermal drug delivery systems and cosmetics. Background technique: [0002] Entering the 21st century, the medical field is facing a new problem: how to correctly deliver the drug to the site to be treated so that the drug effect can be maximized. The beauty industry is also facing the same problem. Why is the effect of some high-tech nanotechnology, first-class raw materials and expensive beauty creams and beauty creams not as obvious after application? Therefore, in recent years, academic research on transdermal drug delivery, transdermal absorption, and sustained release in the medical field has been unprecedentedly active. Over the past ten years of research, it has been found that the immunodeficiency virus (HIV-1) transcriptional activator Tat-mediated protein has a good membrane-penetrating effect. We...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/42A61K47/48A61K8/64A61K9/00A61K9/06A61K9/08A61K9/107A61K9/12A61K38/18A61P17/00A61Q19/00A61K47/64
Inventor 黄亚东项琪苏志坚丁长才张卉姚崇舜
Owner BIOPHARM RES & DEV CENT JINAN
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