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A cell penetrating peptide and transdermal interferon

A technology for penetrating peptides and interferons, applied in the field of biomedicine, to achieve the effect of small molecular weight, improved therapeutic effect, and rapid effect

Active Publication Date: 2022-06-21
SHANGHAI HUAXIN HIGH BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this phospholipid bilayer is essential for cell survival and function, it poses challenges for the exchange of cargo molecules inside and outside the cell

Method used

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  • A cell penetrating peptide and transdermal interferon
  • A cell penetrating peptide and transdermal interferon
  • A cell penetrating peptide and transdermal interferon

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] 1. Screening of CPP

[0054] Using phage in vivo display technology to screen CPPs that can penetrate the skin. The specific method is,

[0055] 1) Apply the phage peptide library on the skin surface of nude mice by transdermal administration;

[0056] 2) After transdermal for a period of time, blood is drawn from the mouse;

[0057] 3) adding Escherichia coli to the blood, allowing the phage to infect the Escherichia coli, and then culturing;

[0058] 4) Finally, count the plaques and calculate the phage titer.

[0059] The first round of screening is performed according to the above steps. Usually, many non-specific phages or phages with weak skin penetration ability will be screened in the first round of screening, and the number is large. Therefore, repeated screening is required. By repeating the above method. After 3-4 rounds of screening, the phages were enriched to a certain purity.

[0060] The polypeptide sequence is then obtained by extracting the phag...

Embodiment 2

[0069] Obtain fusion protein CPP-IFNα-2b

[0070] (1) Construction of engineering bacteria

[0071] The cpp and IFNα-2b gene sequences screened above were spliced ​​together (cpp-IFNα-2b), and the whole gene was synthesized. Among them, the nucleotide sequence of Cpp encodes the amino acid sequence RKTWLRRLARRPQLK, which is connected to the N segment of IFNα-2b. The nucleotide sequence of cpp-IFNα-2b is shown in SEQ ID NO:2, and the amino acid sequence is shown in SEQ ID NO:3.

[0072] The cpp-IFNα-2b gene fragment was constructed into our company's prokaryotic soluble expression vector PHX53, and transformed into competent cells BL21 (DE3).

[0073] (2) Fermentation culture engineering bacteria

[0074] The genetically engineered bacteria obtained above were inoculated into 2×YT medium, and cultured at 37°C in a 15L fermenter;

[0075] Fermentation process: adjust the rotation speed to control the dissolved oxygen value between 50-65%, and use ammonia water to control the...

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Abstract

The invention belongs to the field of biomedicine and discloses a cell penetrating peptide and a transdermal interferon. The cell-penetrating peptide provided by the present invention is a peptide that mediates the delivery of biologically active molecules into cells, and the amino acid sequence of the cell-penetrating peptide includes the sequence shown in SEQ ID NO: 1; wherein, the third position from the N-terminal, and / or the 8th, and / or the amino acid at the 11th position can be replaced by other amino acids. The cell penetrating peptide of the present invention has high transdermal activity, and can be used as a carrier to carry target substances (such as protein, DNA, siRNA, etc.) The transdermal interferon formed later is easier to be absorbed through the skin and exert its effect more quickly.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a cell penetrating peptide and a transdermal interferon. Background technique [0002] Transdermal drug delivery refers to a drug delivery system in which the drug penetrates the stratum corneum of the skin and enters the capillaries to enter the blood circulation after being administered on the skin surface to play a systemic therapeutic role. As a special route of administration, compared with the traditional oral administration and injection, it has its unique advantages. Good efficacy, and reduce individual differences in medication; maintain a constant and effective blood drug concentration, avoid peak and valley phenomenon of blood drug concentration, and reduce toxic side effects; prolong drug action time, reduce the number of administrations, and make it easy for most patients to accept; no pain , so that it is convenient for patients to self-medicate, and can also...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C07K19/00C12N15/11A61K47/64A61K38/21A61P31/12
CPCA61K47/64A61P31/12C07K7/08A61K38/00C07K2319/10
Inventor 李冠英张德宝李海红刘惠
Owner SHANGHAI HUAXIN HIGH BIOTECH
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