阅读说明：本技术 一种含有重组人纤连蛋白肽的组合物 (Composition containing recombinant human fibronectin peptide ) 是由 项琪 黄亚东 陈颖之 薛来武 于 2019-11-08 设计创作，主要内容包括：本发明公开了一种含有重组人纤连蛋白肽的组合物,由活性物质和辅料组成,所述活性物质包括纤连蛋白、溶菌酶、精氨酸/赖氨酸多肽和三肽-1铜。所述纤连蛋白、溶菌酶、精氨酸/赖氨酸多肽和三肽-1铜的重量比为0.01~10：0.01~10：0.01~10：0.01~10。本发明的组合物在促进创面愈合平整、控油保水等方面功效显著。本发明的组合物复配以适当辅料后,可进一步制备成冻干粉剂、纳米微球、纳米脂质体、水剂、凝胶等半固体制剂,作为活性添加剂应用于组织工程、药学以及美容护肤领域。(The invention discloses a composition containing recombinant human fibronectin peptide, which consists of active substances and auxiliary materials, wherein the active substances comprise fibronectin, lysozyme, arginine/lysine polypeptide and tripeptide-1 copper, the weight ratio of the fibronectin to the lysozyme to the arginine/lysine polypeptide to the tripeptide-1 copper is 0.01 ~ 10: 0.01 ~ 10: 0.01 ~ 10: 0.01 ~ 10, and the composition has obvious effects on promoting the smooth healing of wound surfaces, controlling oil and retaining water and the like.)

1. A composition containing recombinant human fibronectin peptide is characterized by consisting of an active substance and an auxiliary material, wherein the active substance comprises fibronectin, lysozyme and arginine/lysine polypeptide.

2. The composition of claim 1, further comprising the tripeptide-1 copper.

3. The composition of claim 2, comprising recombinant human fibronectin peptide wherein the weight ratio of fibronectin, lysozyme, arginine/lysine polypeptide, and tripeptide-1-copper is 0.01 ~ 10: 0.01 ~ 10: 0.01 ~ 10: 0.01 ~ 10.

4. The composition of claim 1, comprising a recombinant human fibronectin peptide having the amino acid sequence of SEQ ID No. 1.

Technical Field

The invention relates to the fields of biomedicine and cosmetics, in particular to a composition containing recombinant human fibronectin peptide (rhFnp).

Background

The main function of the skin is to protect the body from adverse factors in the external environment. In general, the skin barrier can be divided into broad and narrow skin barriers. Skin barriers in a broad sense include physical barriers, pigment barriers, nerve barriers, immune barriers; the skin barrier in the narrow sense refers primarily to the physical barrier function of the epidermis, and in particular the stratum corneum, which is the first natural line of defense of our body against the external environment. When the skin is damaged, if the skin cannot heal in time, a large amount of exudates appear on the surface of a wound, and the regeneration of epidermis and the closure of the wound are hindered, so that the progress of rehabilitation is obviously influenced.

Skin wound healing is the repair process of damaged skin tissue, including inflammatory reactions, cell proliferation, and remodeling of new tissue. Particularly, in the process of repairing chronic wounds such as diabetic ulcers, repeated infection can occur due to the existence of a biofilm formed by bacteria, the wounds are difficult to heal for a long time, if the dosage of the medicine is increased, side effects can be increased, and the difficulty of treatment is increased due to the continuous generation of clinical drug-resistant bacteria. It is therefore a development and trend in the art to develop a product that is effective, non-resistant after long-term use, and has good biocompatibility and safety.

Fibronectin (abbreviated as fibronectin, FN) is a macromolecular protein located on the cell surface and in plasma, and its main function is to exert structural and adhesive effects in the cellular fibrous matrix. Meanwhile, fibronectin can promote cell growth, improve cell adherence rate, enhance cell metabolism level, shorten cell growth time and the like. FN molecules have multiple functional domains, wherein the heparin binding domain binds to various products with heparin affinity, such as proteins, polypeptides, or other compounds, enhancing their activity. On the basis, the invention is based on the compounding of the independently developed recombinant human fibronectin peptide (invention patent application 201910949562.3) and the arginine/lysine polypeptide, and the generated synergistic effect further promotes the proliferation capacity of the arginine/lysine polypeptide on skin fibroblasts and simultaneously improves the tissue repair function.

The first phase of wound healing is the inflammatory phase, in which after a wound has formed, blood clots fill the wound or tissue crevice, and the surrounding tissue undergoes acute inflammation. During the inflammation process, on one hand, harmful substances such as bacteria are accumulated near the wound to cause infection; on the other hand, the increase of vascular permeability causes tissue edema and suppurative lytic destruction, and delays wound healing. Lysozyme (also called muramidase) or N-acetylmuramidase hydrolase, which is an alkaline enzyme that hydrolyzes mucopolysaccharides in pathogenic bacteria. The bacterial lysis is caused by the breakdown of the insoluble mucopolysaccharide of the cell wall into soluble glycopeptides, mainly by breaking the beta-1, 4 glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in the cell wall, resulting in the escape of the contents of the cell wall fission. Lysozyme can also be combined with various acidic substances which induce inflammation to inactivate the acidic substances, enhance the curative effects of antibiotics and other medicines, and improve the mucopolysaccharide metabolism of tissue matrixes, thereby being beneficial to repairing tissues damaged by inflammation. Human lysozyme can activate blood platelet, improve local tissue circulation disorder, and enhance local defense, thereby better resisting bacteria, diminishing inflammation, and relieving pain.

The chronic wound refers to the surface wound caused by various reasons, i.e. the damaged skin tissue cannot be orderly repaired histologically, resulting in a long-lasting non-healing surface, such as other chronic diseases, e.g. diabetes, etc.

After the recombinant fibronectin peptide and the lysozyme are compounded, on one hand, the rhFnp and the arginine/lysine peptide have the synergistic effect of further promoting the skin metabolism and repairing the damaged skin; on the other hand, the lysozyme and arginine/lysine polypeptides in the rhFnp composition can disrupt the biofilm formed by the bacteria at the wound site, thereby inhibiting the bacteria from aggregating. The compound effect is obviously better than that of single use. Meanwhile, the fibronectin used by the product of the invention is a fermentation product extract containing fibronectin obtained by expression through a gene recombination technology, and the problems of low protein yield, high cost and the like are effectively solved.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide a composition containing recombinant human fibronectin, which has the effects of promoting the smooth healing of a wound surface, controlling oil and preserving water.

In order to achieve the purpose, the invention adopts the following technical scheme:

a composition containing recombinant human fibronectin peptide comprises active substances and adjuvants, wherein the active substances include fibronectin, lysozyme and arginine/lysine polypeptide.

Preferably, the composition comprising recombinant human fibronectin peptide further comprises the tripeptide-1 copper.

Preferably, in the above composition comprising recombinant human fibronectin peptide, the weight ratio of fibronectin, lysozyme, arginine/lysine polypeptide, and tripeptide-1 copper is 0.01 ~ 10: 0.01 ~ 10: 0.01 ~ 10: 0.01 ~ 10.

Preferably, in the composition containing a recombinant human fibronectin peptide, the fibronectin is a recombinant human fibronectin peptide (rhFnp), and the amino acid sequence of the fibronectin is represented by SEQ ID No. 1.

Compared with the prior art, the invention has the advantages that the fibronectin is selected to be compounded with the lysozyme and the arginine/lysine polypeptide, the lysozyme is added into the composition, the lysozyme has the functions of antibiosis, antiphlogosis, antivirus and the like, can effectively inhibit the inflammation and is beneficial to the wound repair, the arginine/lysine polypeptide has the anti-wrinkle effect, tripeptide-1 copper is added and other components of the composition can be cooperated to promote the regeneration of epithelial tissues of the skin, the obtained composition has obvious effects on promoting the smooth healing of wound surfaces, controlling oil and retaining water and the like, the more optimized scheme is that the region 12 ~ 14 of the amino acid sequence of the human fibronectin is intercepted, the recombinant human fibronectin peptide rhFnps with small molecular weight and original characteristics of the fibronectin is reserved, the cell growth can be promoted, the cell adherence rate is improved, the cell metabolism level is enhanced, the recombinant human fibronectin peptide rhFrhFnps are adopted, the molecular weight is small, the absorption effect is easy, and the composition can be further prepared into freeze-drying powder, nano-microsphere, nano liposome, water aqua, gel and other active preparations as semi-solid active additives and pharmaceutical additives to be applied to the skin care engineering fields.

Drawings

FIG. 1 shows the adhesion promoting effect of rhFnp composition on Hacat cells.

FIG. 2 shows the pro-proliferative activity of rhFnp combinations on Balb/c-3T3 cells.

Fig. 3 is a graph of the spreading, migration and wound healing effects of rhFnp compositions on Hacat cells.

FIG. 4 shows the interaction assay between rhFnp and arginine/lysine polypeptide.

Fig. 5 shows the bacteriostatic effect of rhFnp composition on bacteria (staphylococcus aureus).

Fig. 6 shows the variation in the use of the composition containing rhFnp (example 3) by subjects; in the figure, reference symbol A represents a comparative example, and B represents an experimental group.

Detailed Description

The present invention is further described with reference to the following drawings, but it should be understood by those skilled in the art that the present invention is not limited to these specific embodiments.