CN110664995B

CN110664995B - Composition containing recombinant human fibronectin peptide

Info

Publication number: CN110664995B
Application number: CN201911084430.5A
Authority: CN
Inventors: 项琪; 黄亚东; 陈颖之; 薛来武
Original assignee: Guangzhou Jinan University Medical Biotechnology Research And Development Center Co ltd; Guangzhou Jida Biotechnology Co ltd
Current assignee: Guangzhou Jinan University Medical Biotechnology Research And Development Center Co ltd; Guangzhou Jida Biotechnology Co ltd
Priority date: 2019-11-08
Filing date: 2019-11-08
Publication date: 2022-01-14
Anticipated expiration: 2039-11-08
Also published as: CN110664995A

Abstract

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 to 10. The composition has remarkable effects on promoting smooth healing of wound surfaces, controlling oil and retaining water and the like. After being compounded with proper auxiliary materials, the composition can be further prepared into semisolid preparations such as freeze-drying powder, nano microspheres, nano liposomes, water aqua, gel and the like, and can be used as an active additive to be applied to the fields of tissue engineering, pharmacy and beauty and skin care.

Description

Composition containing recombinant human fibronectin peptide

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 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 to 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 following beneficial effects: the invention selects fibronectin, lysozyme and arginine/lysine polypeptide to be compounded, and the three have synergistic action; the lysozyme is added into the composition, has the functions of antibiosis, antiphlogosis, antivirus and the like, can effectively inhibit the generation of inflammation, and is beneficial to the repair of wounds. The arginine/lysine polypeptide has anti-wrinkle effect. The tripeptide-1 copper and other components of the synergistic composition are added to promote the regeneration of skin epithelial tissue. The obtained composition has obvious effects on promoting smooth healing of wound surfaces, controlling oil and retaining water and the like. The more optimized scheme is that a region 12-14 of the amino acid sequence of human fibronectin is intercepted, and the recombinant human fibronectin peptide rhFnp which has small molecular weight and retains the original characteristics of fibronectin can promote cell growth, improve the cell adherence rate and enhance the cell metabolism level. The invention adopts the rhFnp segment of the recombinant human fibronectin peptide, has small molecular weight and is easy to absorb and play a role. After being compounded with proper auxiliary materials, the composition can be further prepared into semisolid preparations such as freeze-drying powder, nano microspheres, nano liposomes, water aqua, gel and the like, and can be used as an active additive to be applied to the fields of tissue engineering, pharmacy and beauty and skin care.

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.

Example 1: preparation of fibronectin-containing compositions

A composition containing fibronectin, wherein the mass concentration of each component is shown in Table 1:

TABLE 1

Components	Mass concentration ratio
		Fibronectin	40μg/mL
Lysozyme	35 μg/mL
		Arginine/lysine polypeptides	5μg/mL
Tripeptide-1 copper	500μg/mL
		Water (W)	1mL

Mixing the above materials at a certain proportion, and swelling or dissolving in water.

Example 2: preparation of compositions containing rhFnp

A composition containing rhFnp and lysozyme comprises the following components in mass concentration as shown in Table 2:

TABLE 2

Components	Mass concentration ratio
		rhFnp	40μg/mL
Lysozyme	35 μg/mL
		Arginine/lysine polypeptides	5μg/mL
Tripeptide-1 copper	500μg/mL
		Water (W)	1mL

Example 3: preparation of compositions containing rhFnp

A composition containing rhFnp and lysozyme comprises the following components in mass concentration as shown in Table 3:

TABLE 3

Components	Mass concentration ratio
		rhFnp	75μg/mL
Lysozyme	35 μg/mL
		Arginine/lysine polypeptides	5μg/mL
Tripeptide-1 copper	500μg/mL
		Water (W)	1mL

Example 4: preparation of compositions containing rhFnp

A composition containing rhFnp and lysozyme comprises the following components in mass concentration as shown in Table 4:

TABLE 4

Components	Mass concentration ratio
		rhFnp	100μg/mL
Lysozyme	35 μg/mL
		Arginine/lysine polypeptides	5μg/mL
Tripeptide-1 copper	500μg/mL
		Water (W)	1mL

Comparative example 1: preparation of composition containing lysozyme

A lysozyme composition for bacteriostasis and anti-inflammation comprises the following components in percentage by mass as shown in Table 5:

TABLE 5

Components	Mass concentration ratio
		Lysozyme	35 μg/mL
Arginine/lysine polypeptides	5μg/mL
		Tripeptide-1 copper	500μg/mL
Water (W)	1mL

Example 5: preparation of hydrogel containing rhFnp composition

Weighing a certain amount of dried Chitosan (CS) powder, adding 1-3% of acetic acid solution to prepare 1-2% (w/v) of CS solution, adding the composition in example 3 into the CS solution, uniformly mixing, adding 56-58% of beta-sodium glycerophosphate aqueous solution, stirring, uniformly mixing, and incubating in water bath at 37 ℃ for a period of time to form the hydrogel containing the rhFnp composition.

Example 6: preparation of rhFnp-containing composition nano-microsphere

Weighing a certain amount of dried Chitosan (CS) powder, adding the CS powder into 1-3% acetic acid solution to prepare 1-2% (w/v) CS solution serving as a water phase, and storing the CS solution at 4 ℃ for later use. Preparing a certain amount of span80 and liquid paraffin with the ratio of 1:30-35 into oil phase. After being mixed by a magnetic stirrer, 1mL of the aqueous phase is dropwise added into 25mL of the oil phase by a 5mL syringe while stirring, and the stirring speed is 700 r/min. After stirring for 2h, 1.5mL of 25% glutaraldehyde crosslinker were added in three portions and stirring was continued for 4 h. Standing for 30min, removing supernatant, washing with petroleum ether, isopropanol, ethanol and ultrapure water, and freeze-drying to obtain chitosan microsphere powder. And adding the solution containing the rhFnp composition into 20mg of chitosan microspheres, oscillating by a shaking table to adsorb the rhFnp composition onto the chitosan microspheres, and centrifuging to remove the supernatant to obtain the nano microspheres containing the rhFnp composition.

Example 7: preparation of rhFnp-containing composition nano-microsphere

Weighing a certain amount of dried Chitosan (CS) powder, adding the CS powder into 1-3% acetic acid solution to prepare 1-2% (w/v) CS solution, then adding a certain amount of solution containing the rhFnp composition, uniformly mixing the solution to be used as water phase, and storing the water phase at 4 ℃ for later use. Preparing a certain amount of span80 and liquid paraffin with the ratio of 1:30-35 into oil phase. After being mixed by a magnetic stirrer, 1mL of the aqueous phase is dropwise added into 25mL of the oil phase by a 5mL syringe while stirring, and the stirring speed is 700 r/min. After stirring for 2h, 1.5mL of 25% glutaraldehyde crosslinker were added in three portions and stirring was continued for 4 h. Standing for 30min, removing supernatant, washing with isopropanol, ethanol and ultrapure water, and freeze-drying to obtain the nano-microsphere containing the rhFnp composition.

Example 8: preparation of liposomes containing rhFnp compositions

Weighing a certain amount of lecithin and cholesterol, mixing according to a ratio of 3-5: 1, adding ethanol, mixing uniformly, and adding into a rotary evaporator. After the ethanol is volatilized, the solution containing the rhFnp composition is hydrated and stands overnight, and then the solution is homogenized by a homogenizer and filtered by a 0.22 mu m filter membrane to obtain the nano liposome containing the rhFnp composition.

Example 9: assay for cell adhesion Activity of rhFnp compositions

Hacat cells were cultured in DMEM containing 10% FBS at 37 ℃ with CO₂The concentration is 5%; firstly, washing the cells once by PBS, then adding 0.25% pancreatin solution for digestion, and centrifugally collecting the cells; resuspending with DMEM and controlling the cell density at 6.2X 10⁴one/mL, the cell suspensions were inoculated into the culture dishes plated with rhFnp protein, example 2, example 3 and example 4, respectively, and the cell density was controlled at 1.5X 10⁴one/mL. Culturing at 37 deg.C for 5 hr, maintaining CO₂The concentration is 5%; washing the non-adherent cells with PBS; the number of cells in each group was counted under a phase contrast microscope and compared by the MTT method. The results are shown in FIG. 1 and show that: the number of cells in the rhFnp composition administered at the intermediate concentration and the positive control group was higher than those in the control group and the rhFnp within the concentration range examined, with the rhFnp composition of example 3 having the highest number of cells. The results indicate that the rhFnp composition has activity in promoting cell adhesion.

Example 10: assay for the proliferative Activity of rhFnp compositions

Balb/c-3T3 cells in logarithmic growth phase were routinely cultured in 1640 medium containing 10% FBS (purchased from Gibco, USA) to 80% -90% confluence, digested with 0.25% pancreatin, and cultured at 2.0X 10⁵The number of cells per mL was plated in 96-well plates, and after 24h of culture, the plates were cultured in 1640 medium containing 0.4% serum, and after 24h of culture medium was aspirated, rhFnp, the rhFnp compositions of example 2, example 3 and example 4 (all samples were diluted to 0.5mg/mL with PBS), 3 duplicate wells were placed in each group, and six duplicate wells were left in blank groups. After administration, the cells were placed in an incubator and incubated for another 48 hours with 20. mu.L MTT (0.5mg/mL) and for 4 hours, the wells were aspirated and 100. mu.L DMSO was added, and the absorbance OD was measured at a wavelength of 570 nm/630nm after shaking. The experiment can detect the proliferation promoting activity of the rhFnp composition on Balb/c-3T3 cells, and the result is shown in FIG. 2, and the result shows that the cell proliferation rate of the rhFnp composition administration group is higher than that of rhFnp in the investigation range, which indicates that the rhFnp composition has the activity of promoting the proliferation of Balb/c-3T3 cells.

Example 11: scratch test of rhFnp composition

Balb/c-3T3 cells in logarithmic growth phase were routinely cultured in 1640 medium containing 10% FBS (from Gibco, USA) to 80% -90% confluence, digested with 0.25% trypsin and plated into 12-well plates, preferably in such amounts that the plate bottom was confluent after adherence (when the amount is small, the cells were cultured for a period of time to confluent plate bottom to ensure that the cells reached a 100% confluent density). After the cells grow to be full of the bottom of the plate, a 100-microliter gun head is used for being vertical to the orifice plate, cell scratches are made at the same positions of the back surface of the plate where the scratches are made, and the width of each scratch is ensured to be consistent as much as possible. The cell culture was aspirated, the well plate was rinsed three times with PBS, and cell debris generated by scratching was washed away. The culture medium containing rhFnp protein, example 2, example 3 and example 4, which contains 1% serum, was added and recorded by photography. The culture plate is put into an incubator to be cultured for 24h and taken out to be photographed.

Example 12: interaction assay of rhFnp with arginine/lysine polypeptide

Balb/c-3T3 cells in logarithmic growth phase were routinely cultured in 1640 medium containing 10% FBS (purchased from Gibco, USA) to 80% -90% confluence, digested with 0.25% pancreatin, and cultured at 2.0X 10⁵Inoculating the number of cells per mL into a 96-well plate, culturing for 24h, then changing a 1640 culture medium containing 0.4% serum for culturing, sucking out the culture medium after 24h, then respectively adding the 1640 culture mediums containing arginine/lysine polypeptide, rhFnp and arginine/lysine polypeptide in the mass concentration ratio of 2:1, 1:1 and 1:2 for culturing, wherein each concentration is provided with 3 multiple wells, and the blank group is six multiple wells. After administration, the cells were placed in an incubator and incubated for another 48 hours with 20. mu.L MTT (0.5mg/mL) and for 4 hours, the wells were aspirated and 100. mu.L DMSO was added, and the absorbance OD was measured at a wavelength of 570 nm/630nm after shaking. The experiment can detect the proliferation promoting activity of the rhFnp composition on Balb/c-3T3 cells, and the result is shown in FIG. 4, and the result shows that in the investigated range, when the mass concentration ratio of the rhFnp to the arginine/lysine polypeptide is 1:1, the cell proliferation rate is higher than that of the single arginine/lysine polypeptide, and the activity of the interaction of the rhFnp and the arginine/lysine polypeptide for promoting the proliferation of Balb/c-3T3 cells is shown.

Example 13: bacteriostatic assay of rhFnp composition (example 3) against Staphylococcus aureus

Staphylococcus aureus (purchased from Guangdong province center for culture Collection of microorganisms) was anaerobically cultured in an anaerobic tank at 37 ℃ for 48h (broth culture medium, purchased from Wejia Bio Inc., Guangzhou), diluted to 1X10 with sterile physiological saline⁶CF μm/ml. The rhFnp composition of example 3 was prepared in broth medium at different concentrations and 0.1mL of the broth was taken to sterile test tubes of the rhFnp composition of example 3 at different concentrations in a volume of 1 mL. Culturing at 37 deg.C for 48h in anaerobic tank, and indicating oxygen content in anaerobic tank with oxygen indicator (when oxygen concentration is greater than 0.5%, the oxygen indicator changes from purple or purple red to blue). The presence or absence of turbidity was observed, and the Minimum Inhibitory Concentration (MIC) was determined as the lowest concentration at which turbidity did not occur.

The results are shown in FIG. 5. In FIG. 4, 1-6 are blank control (without Staphylococcus aureus), comparative example 1, negative control (without rhFnp composition), and example 3 rhFnp composition with total protein mass concentration of 500, 250, 125 μ g/ml. The results showed that turbidity occurred in the test tube medium at 250. mu.g/ml, indicating that the rhFnp composition prepared in example 3 had a Minimum Inhibitory Concentration (MIC) of 500. mu.g/ml against Staphylococcus aureus.

Example 14: rhFnp composition (example 3) repair skin damage test

The effective number of the subjects is 18 cases; 18-45 years old; the subject has no flaws in arm; the tested part does not participate in skin treatment, beauty treatment or other skin tests for nearly three months; no chemical exfoliation or laser treatment was performed in the last half year and informed consent was signed.

The intact forearm of the subject was selected, minimally invasive (depth 2.0 mm) was performed with an electric microneedle, and left at a fixed position at point 50 for 1 second each time with a slight downward force to achieve uniform redness and slight bleeding, so that the lesions at all points were as uniform as possible.

Use of the composition:

a: the microneedles are not treated thereafter,

b: post-microneedle application of the composition of example 3 (4 times per day);

b, smearing corresponding solutions at corresponding time points (8 points, 12 points, 16 points and 21 points) within 6 days after operation, and detecting CK every other day.

The formal test shows that the people can not drink water and beverage after sitting still for 30min in a room (the temperature is 20-22 ℃, and the humidity is 40-60%) meeting the standard.

The test results of the percutaneous water loss rate (TEWL) before and after the use of the subjects were shown in table 6, and the test results of the red pigment at the microneedles were shown in table 7.

TABLE 6

Point in time	A	B
			D1	24.93±6.95	25.5±9.38
D3	13.18±3.94****	10.88±2.31****
			D6	12.35±3.89****	10.72±3.80****
D1VS. D3 percent	-47.13%	-57.33%
			D1VS. D6 percent	-50.48%	-57.97%
The difference D3-D1	-11.75	-14.62
			The difference D6-D1	-12.58	-14.78

VS.D1 ***P<0.001, ****P<0.0001

VS. A ^# P<0.05, ^## P<0.01

TABLE 7

Point in time	A	B
			D1	398.61±74.74	368.31±71.16
D3	342.17±64.67*	300.69±58.53**
			D6	288.09±47.31****	259.74±51.91****
D1VSD3 percent	-14.16%	-18.36%
			D1VSD6 percent	-27.73%	-29.48%
The difference D3-D1	-56.44	-67.62
			The difference D6-D1	-110.52	-108.57

VS.D1 ***P<0.001, ****P<0.0001

VS. A ^# P<0.05, ^## P<0.01

As shown in fig. 6, it can be seen that the TEWL value decreased after the microneedles of the subjects in the comparative example and using the composition of example 3, and the decrease in TEWL was significant at Day 3 in the experimental group, demonstrating that example 3 can moisturize and lock water; the decrease in red pigment is very marked after the experimental group used the composition 6d of example 3.

Sequence listing

<110> Guangzhou city and Biotechnology, Inc.; medical biotechnology research and development center of Guangzhou river-south university

<120> a composition comprising a recombinant human fibronectin peptide

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 281

<212> PRT

<213> Artificial sequence ()

<400> 1

Gly Thr His Met His His His His His His Pro Ala Pro Thr Asp Leu

1 5 10 15

Lys Phe Thr Gln Val Thr Pro Thr Ser Leu Ser Ala Gln Trp Thr Pro

20 25 30

Pro Asn Val Gln Leu Thr Gly Tyr Arg Val Arg Val Thr Pro Lys Glu

35 40 45

Lys Thr Gly Pro Met Phe Val Ala Gln Cys His Pro Phe Ser Cys Ala

50 55 60

Arg Lys Phe His Gly Leu Met Val Asp Cys Thr Cys Leu Val Ser Val

65 70 75 80

Tyr Ala Leu Lys Asp Thr Leu Thr Ser Arg Pro Ala Gln Gly Val Val

85 90 95

Thr Thr Leu Glu Asn Val Ser Pro Pro Arg Arg Ala Arg Val Thr Asp

100 105 110

Ala Thr Glu Thr Thr Ile Thr Ile Ser Trp Arg Thr Lys Thr Glu Thr

115 120 125

Ile Thr Gly Phe Gln Val Asp Ala Val Pro Ala Asn Gly Gln Thr Pro

130 135 140

Ile Gln Arg Thr Ile Lys Pro Asp Val Arg Ser Tyr Thr Ile Thr Gly

145 150 155 160

Leu Gln Pro Gly Thr Asp Tyr Thr Val Gly Met Gln Trp Leu Asn Asp

165 170 175

Asn Ala Arg Ser Ser Pro Val Val Ile Asp Ala Ser Thr Ala Ile Asp

180 185 190

Ala Pro Ser Asn Leu Arg Phe Leu Ala Thr Thr Pro Asn Ser Leu Leu

195 200 205

Arg Tyr His Gln Arg Thr Asn Thr Asn Val Met Pro Trp Phe Val Pro

210 215 220

Val Thr Glu Gly Ala Glu Ala Leu Thr Ala Arg Val Asn Leu Asn Leu

225 230 235 240

Pro Gly Val Thr Glu Ala Thr Ile Thr Gly Leu Glu Pro Gly Thr Glu

245 250 255

Tyr Thr Ile Tyr Val Ile Ala Leu Lys Asn Asn Gln Lys Ser Glu Pro

260 265 270

Leu Ile Gly Arg Lys Lys Thr Lys Leu

275 280

Claims

1. A composition containing recombinant human fibronectin peptide is characterized by comprising an active substance and an auxiliary material, wherein the active substance comprises fibronectin, lysozyme and arginine/lysine polypeptide; also contains tripeptide-1 copper;

580-615 mu g of active substance is contained in each ml of water, wherein the weight ratio of the fibronectin, the lysozyme, the arginine/lysine polypeptide and the tripeptide-1 copper is 2.5-10: 0.7-10: 0.1-10: 10;

the fibronectin is recombinant human fibronectin peptide, and the amino acid sequence of the fibronectin is shown in SEQ ID NO. 1.