CN-121471377-B - High-transdermal recombinant humanized 17-type collagen and application thereof

Abstract

The invention belongs to the technical field of genetic engineering, and relates to high-transdermal recombinant humanized 17-type collagen and application thereof. The collagen peptide consists of a penetrating peptide section and a collagen section, wherein the collagen section is an amino acid sequence shown as SEQ ID NO. 1, and the penetrating peptide section is penetrating peptide SPACE, TAT or ANTP. The high-transdermal recombinant humanized 17-type collagen provided by the invention not only has higher skin permeation efficiency, but also has multiple biological functions of barrier repair, tightening, wrinkle resistance and the like, and is beneficial to the high-efficiency application in the fields of cosmetics, medicines and the like.

Inventors

• YUAN FENGJIAO
• LI ZHENAI
• ZHANG WENWEN
• ZHENG DEQIANG
• LI HAIJUN
• MAO ZIHAN
• LU JIE
• LI WENHAO
• LIU YUJIE
• Zou Juhua
• ZHANG YINGHUA

Assignees

• 山东福瑞达医药集团有限公司
• 山东福瑞达生物科技有限公司

Dates

Publication Date

20260508

Application Date

20260108

Claims (9)

1. 1. The high-transdermal recombinant humanized type 17 collagen is characterized by comprising a penetrating peptide section and a collagen section, wherein the collagen section is an amino acid sequence shown as SEQ ID NO. 1, and the penetrating peptide section is a penetrating peptide TAT; the membrane penetrating peptide segment is modified at the N end of the collagen segment; or the amino acid sequence of the high-transdermal recombinant humanized type 17 collagen is shown as SEQ ID NO. 5.
2. 2. A nucleic acid molecule encoding the highly transdermal recombinant humanized type 17 collagen of claim 1.
3. 3. The nucleic acid molecule of claim 2, wherein the DNA sequence of said nucleic acid molecule is set forth in SEQ ID No. 9.
4. 4. A recombinant vector comprising the nucleic acid molecule of claim 2 or 3.
5. 5. The recombinant vector according to claim 4, wherein the vector used in the recombinant vector is pPIC9K.
6. 6. A recombinant engineering bacterium comprising the nucleic acid molecule according to claim 2 or 3 or the recombinant vector according to claim 4 or 5.
7. 7. A method for preparing high-transdermal recombinant humanized type 17 collagen is characterized by comprising the steps of culturing the recombinant engineering bacterium according to claim 6 so as to express the high-transdermal recombinant humanized type 17 collagen and separating the high-transdermal recombinant humanized type 17 collagen.
8. 8. The method of claim 7, comprising the steps of: (1) Activating the recombinant engineering bacteria; (2) Fermenting the activated thalli until the wet thalli amount reaches 250-400 g/L, stopping feeding, and adding methanol in stages to induce 48-96 h to obtain fermentation liquor; (3) Purifying the obtained collagen by using a multimode weak cation exchange chromatographic packing material to obtain the high-transdermal recombinant humanized XVII type collagen.
9. 9. Use of a highly transdermal recombinant humanized type 17 collagen according to claim 1 or a nucleic acid molecule according to claim 2 or 3 or a recombinant vector according to claim 4 or 5 or a recombinant engineering bacterium according to claim 6 for the preparation of a product for improving skin aging or skin repair.

Description

High-transdermal recombinant humanized 17-type collagen and application thereof Technical Field The invention belongs to the technical field of genetic engineering, and relates to high-transdermal recombinant humanized 17 (XVII) type collagen and application thereof. Background The disclosure of this background section is only intended to increase the understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art already known to those of ordinary skill in the art. 17 Type (XVII) collagen (COL 17) is a core functional protein present in the basal layer of the epidermis of the skin. COL17 is a core component for maintaining an 'anchoring connection' between epidermis and dermis, and is important for stabilizing a basement membrane and preventing epidermis from falling off, and meanwhile, COL17 can regulate and control the activity of epidermal stem cells, promote the ordered replacement of epidermal cells and maintain the continuous repair capability of skin barriers. Therefore, COL17 is considered as a "daemon of skin basement membrane integrity" and an "anti-aging key target". However, with the age (for example, the COL17 synthesis rate of a human body is reduced by 1% -2% per year after the age of 30 years), the ultraviolet irradiation (UVB can induce the COL17 degrading enzyme activity to be improved by 3-5 times), the environmental pollution, the bad living habit and other factors affect the effect, the content of COL17 in the skin can be gradually reduced, the functions can be obviously declined, on one hand, the anchoring structure of the basement membrane is loose, the connection gap between the epidermis and the dermis is enlarged, the skin elasticity is reduced, the relaxation and sagging are caused, on the other hand, the updating efficiency of the epidermal stem cells is reduced, the skin barrier repairing capability is reduced, and the aging characteristics such as dryness, sensitivity, wrinkles and the like are displayed. To improve the skin aging problem, exogenous supplementation of COL17 has become a central research direction in the fields of cosmetics and dermatologic medicine repair. Compared with animal-derived COL17, the recombinant human-derived 17-type collagen has the remarkable advantages of low immunogenicity, high biocompatibility, definite functional targeting and the like, and is considered as an ideal material for replacing the traditional collagen. However, a central technical bottleneck currently limiting the application of recombinant human type 17 collagen is the extremely low transdermal absorption efficiency. The reason is that the molecular weight of the recombinant expressed protein is generally higher (far greater than the 500Da penetration threshold of the skin stratum corneum gap), the molecular structure is in a linear rigid conformation, the molecular structure is difficult to pass through the structural barrier of the skin stratum corneum, the transdermal absorption rate is less than 5%, and most of active ingredients only stay on the skin surface layer and cannot effectively reach the target site of the epidermis basal layer to exert the repairing and anti-aging effects. Therefore, there is a need to develop a technology that is efficient, safe, and can promote the transdermal penetration of COL17, thereby promoting efficient application of COL17 in the fields of cosmetics and medicine. Disclosure of Invention In order to solve the defects of the prior art, the invention aims to provide the high-transdermal recombinant humanized 17-type collagen and the application thereof, and the high-transdermal recombinant humanized 17-type collagen provided by the invention not only has higher skin permeation efficiency, but also has multiple biological functions such as barrier repair, tightening, wrinkle resistance and the like, and is beneficial to the high-efficiency application in the fields of cosmetics, medicines and the like. In order to achieve the above purpose, the technical scheme of the invention is as follows: In a first aspect, the high-transdermal recombinant humanized type 17 collagen is composed of a penetrating peptide segment and a collagen segment, wherein the collagen segment is an amino acid sequence shown as SEQ ID NO. 1, and the penetrating peptide segment is a penetrating peptide SPACE, TAT or ANTP. The collagen provided by the invention contains a plurality of KGD active motifs by designing the amino acid sequence of the collagen section, can mediate adhesion and migration of keratinocytes, and is directly related to hair follicle stem cell stabilization and skin basement membrane repair. In designing a collagen segment, the structural integrity, the expression efficiency and the feasibility of subsequent delivery are considered, if the functional domain is too short, a stable triple helix structure and enough active sites are diffic