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Peptides and their preparation and use

A host cell and sequence technology, applied in the field of genetic engineering, can solve the problems of limited wide use, high cost, limited chemical synthesis of hexapeptides, etc., and achieve the effects of low production cost, correct detection molecular weight, and high product purity.

Active Publication Date: 2020-08-04
SHANXI JINBO BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current preparation methods of hexapeptides are still limited to chemical synthesis, and its high cost limits the widespread use of such products

Method used

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  • Peptides and their preparation and use
  • Peptides and their preparation and use
  • Peptides and their preparation and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1: Construction and expression of pET-32a-E8-8 gene expression vector

[0063] (1) Construction of Escherichia coli Genetic Engineering Bacteria

[0064] 1.1. The expression efficiency of the polypeptide can be significantly improved by expressing the E8 polypeptide gene in tandem. In this embodiment, n=8, that is, E8-8 with 8 repeats, whose amino acid sequence is GEEMQRRNGEEMQRRNGEEMQRRNGEEMQRRNGEEMQRRNGEEMQRRNGEEMQRRNGEEMQRRN.

[0065] 1.2.根据E8-8的氨基酸序列,优化选择其大肠杆菌偏好的密码子基因,也就是ggtgaggaaatgcagcgccgcaatggcgaagaaatgcagcgccgcaatggcgaagagatgcaacgccgcaatggcgaggagatgcagcgccgcaatggcgaggaaatgcagcgccgcaatggcgaggagatgcaacgccgcaatggcgaggaaatgcagcgccgcaacggcgaagaaatgcaacgccgcaat(SEQ ID No.5),为了后续多肽酶切的需求,这个基因的5'端加入AAT(羟胺 Restriction site), TAA (stop codon) is added to the 3' end, and the complete gene is aat ggtgaggaaatgcagcgccgcaatggcgaagaaatgcagcgccgcaatggcgaagagatgcaacgccgcaatggcgaggagatgcagcgccgcaatggcgaggaaatgcagcgccgcaatggcgaggagatgcaacgccgcaatggcgaggaaatgcagcgccgcaac...

Embodiment 2

[0078] Embodiment 2: human trial of anti-wrinkle short peptide E8

[0079] Sterilize anti-wrinkle short peptide E8 with a 0.22 μm filter membrane, dilute it with purified water to 40 μg / mL, and store it in the refrigerator for later use.

[0080] 40 volunteers (20 males, 20 females, aged 30-45 years old) were randomly divided into two groups A and B, 20 each (10 males, 10 females). Try 7ml of 40μg / mL anti-wrinkle short peptide E8 on wrinkles such as eye lines and lip lines. Try 7ml of pure water, measure the depth of wrinkles before the experiment, and stop using other anti-wrinkle cosmetics during the experiment, and measure the depth of wrinkles after 15 days of continuous use.

[0081] Method reference: "E Berard Esca et al. (Italian), translated by Zheng Defang. Clinical observation and instrumental evaluation of the effectiveness of anti-wrinkle cosmetics on skin relief and photoaging skin (English) [J]. Daily Chemical Science, 1998 No. 6: 24-26" (which is incorporated ...

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Abstract

The present invention relates to peptides, methods for their preparation and uses. A peptide comprising the sequence shown in SEQ ID No. 1 or a sequence in which one, two or several amino acids are substituted, deleted, inserted and / or added in the sequence shown in SEQ ID No. 1, said peptide showing inhibition of neuronal Cell viability or wrinkle-removing activity. The present invention also provides peptide-encoding nucleotide sequences, vectors containing the nucleotide sequences, and hosts containing the vectors. The peptide of the present invention can be prepared on a large scale through biological fermentation and can be used in anti-wrinkle skin products.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a peptide and its preparation method and application. Background technique: [0002] Skin wrinkle removal is a good wish of human beings to keep youth forever. Human studies have confirmed that wrinkles are associated with overstimulation of facial muscle fibers. Therefore, it is theoretically possible to reduce wrinkles by directly reducing muscle contraction. Muscle contraction is regulated by neurotransmitters, and the SNARE complex is the source of signals transmitted by neurotransmitters. Botulinum toxin, which is widely used in the market at present, inhibits the activity of nerve cells by inhibiting the function of the SNARE complex, thereby achieving the purpose of removing wrinkles. However, the side effects of botulinum toxin are very huge. Human beings have been studying the functional analogues of botulinum toxin, trying to minimize their side effects whi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C12N15/11C12N15/70C12N1/21A61K8/64A61Q19/08C12R1/19
CPCA61K8/64A61Q19/08C07K7/06C12N15/70
Inventor 杨霞
Owner SHANXI JINBO BIO PHARMA CO LTD
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