CN1640491A - A kind of compound biological preparation of recombinant human epidermal growth factor and its application - Google Patents

Abstract

The invention discloses a recombinant human epidermal growth factor compound biological preparation and application thereof, and the recombinant human epidermal growth factor compound biological preparation provided by the invention comprises 2-10 ng/ml of recombinant human epidermal growth factor; 0-20% fetal bovine serum; 2-10% D-glucose; 0-10 mug/ml insulin; hydrocortisone 0-5 μ g/ml; 0-100 mug/ml ampicillin and 0-100 IU/ml streptomycin sulfate, and the solvent is multi-distilled water. The recombinant human epidermal growth factor compound biological preparation can be used for preparing medicines for treating ulcer, fester, wound and burn; can also be used for producing anti-aging cosmetics and beauty products, so that the skin is kept delicate, wrinkles are reduced, color spots are dark and even disappear, and the skin is finally glossy and elastic; can also be used for preparing medicaments for treating the traumatic ophthalmic diseases.

Description

A kind of compound biological preparation of recombinant human epidermal growth factor and its application

technical field

The invention relates to a biological preparation composition and its application, in particular to a recombinant human epidermal growth factor compound biological preparation and its application.

Background technique

In 1960, Cohen and Levi Montalcin found that when the nerve growth factor NGF was purified in the submandibular gland of mice, injection of a component without nerve growth factor into newborn mice could cause early eyelid opening, early incisor eruption, hair growth inhibition and Growth retardation. In 1962, Cohen reported that this heat-stable and acid-stable protein can promote the development of eyelids and incisors. Histological studies have shown that early eyelid opening is due to a characteristic increase in epidermal thickness and keratinization. Studies on the culture of skin or epidermal tissue blocks in vitro have confirmed that this protein has a direct effect on epidermal growth, so it was named epidermal growth factor (EGF). After decades of research, EGF has been found to be a family of proteins. Brown's research team has shown through random studies that recombinant human epidermal growth factor (rhEGF) has a significant effect on promoting wound healing. Thornton's research group and other research groups found that rhEGF can significantly promote the healing of second-degree burns and superficial first-degree wounds in rats (P<0.01), and it can also promote healing in the donor area (P<0.01); and the promoting effect of rhEGF It is mainly manifested in the later stage of wound healing. In the early stage of wound healing, the superficial necrotic tissue tightly covers the wound surface, so that rhEGF applied locally cannot penetrate and act on the remaining dermis and epithelial cells of the skin appendages. The growth-promoting effect can only be exerted after the island is established. Studies have shown that during the clinical use of rhEGF, no abnormalities were found in the liver, kidney function and blood routine of the users, no systemic and local allergic reactions, and no toxic and side effects occurred. After topical application of rhEGF, wound exudation was significantly and rapidly reduced, which may be related to factors such as rhEGF promoting rapid epithelial regeneration.

EGF can promote the mitosis of a variety of epithelial cells, and at the same time stimulate the proliferation of various mesenchymal cells, including corneal endothelial cells, gastrointestinal mucosal cells, vascular endothelial cells, vascular smooth muscle cells, fibroblasts and chondrocytes. The growth of the epidermis provides the required base material. In addition to promoting proliferation, EGF can also promote the migration of certain cells such as corneal endothelial cells and vascular endothelial cells, which is necessary for wound repair. EGF can promote the repair of corneal and skin wound epithelial cells and gastrointestinal mucosal ulcer wounds, promote the proliferation of epidermal cells, and promote the deposition of collagen by stimulating the proliferation of fibroblasts and other cells. EGF can non-sustainably improve the proliferation ability of fibroblasts in a certain period of time. EGF can greatly promote the growth of dermal fibroblasts, promote the expression of Cyclin D1 and CDk4, shorten the cell cycle, and enhance the proliferation ability. Savage found in 1972 that EGF can promote corneal epithelial proliferation, and many scholars have also confirmed this by using animal corneal injury models such as corneal incision, corneal scraping, corneal epithelial resection, and alkali burn. In addition, in some non-dystrophic diseases of corneal epithelium, it is also found that EGF can promote the repair of corneal damage. In 1989, Brown et al. performed skin transplantation on 12 patients undergoing plastic surgery and burns, and found that the use of EGF could significantly promote wound healing. In addition, EGF also has obvious therapeutic effect on gastric ulcer.

There are EGF receptors on gastric parietal cells and duodenal epithelial cells. EGF directly enters the digestive tract through saliva or is directly secreted into the small intestine by duodenal glands to exert a biological effect on the mucosa of the digestive tract. EGF is an important protective factor of gastric mucosa, which plays a role by binding to the specific receptor EGFR on gastrointestinal mucosal cells. The level of EGF in gastric juice is one of the important indicators for evaluating the functional maturity of regenerated mucosa in the quality of ulcer healing. Animal experiments have shown that intragastric perfusion of EGF in mice can prevent gastric ulcers induced by aspirin, and exogenous EGF can significantly inhibit drug-induced gastric and duodenal ulcers.

Existing studies have shown that the EGF content in gastric juice of patients with peptic ulcer is significantly lower than that of normal people, suggesting that the reduction of EGF may have a certain relationship with the occurrence of gastric ulcer; may work. At present, according to experiments, it is speculated that the possible mechanism of EGF involved in the repair of peptic ulcer is ①activating ornithine decarboxylase (ODC), increasing mucosal DNA synthesis; ②inhibiting gastric acid secretion; ③promoting the synthesis and secretion of gastric mucus glycoprotein; ④increasing Gastric mucosal blood flow. In clinical work, the possibility of ulcer occurrence can be predicted by dynamically monitoring the EGF content of gastric juice, and the efficacy of drugs can also be observed during treatment.

EGF is a powerful factor that promotes cell division, differentiation, and proliferation. It is known that it can promote proliferation and stimulate DNA synthesis in various tissues. It can promote cell growth by acting on specific receptors on the target cell membrane. growth activity. In vitro studies have shown that EGF can actively stimulate the proliferation of skin epithelial cells and fibroblasts, promote the formation of new blood vessels and the synthesis of nucleic acids, proteins and hydroxyproline. Animal experiments have confirmed that a very small amount of recombinant human epidermal growth factor (rhEGF) can play a role, but increasing the dose will affect its effect because the number of EGF receptors on the cell membrane is "negatively regulated" by rhEGF.

Some data show that EGF regulates the behavior of cells through the remodeling of the extracellular matrix (ECM), or the result of ECM remodeling is the release of EGF to initiate changes in cell phenotype. ECM can separate EGF-binding proteins, namely related Theory of regulation of growth factor activity. With the development of cell biology and molecular biology techniques, the role of cytokines in the process of wound repair is gradually being paid attention to.

Insulin is secreted by pancreatic beta cells. Proinsulin contains 86 amino acids with a molecular weight of about 6,000. Insulin consists of 51 amino acids. The structure of different animals is slightly different, but it does not affect its physiological activity. Insulin can promote the uptake of glucose, amino acids and ions by cells, enhance the phosphorylation of tyrosine and tryptophan in cells, adjust the activity of regulatory enzymes, stimulate or inhibit the transcription of specific genes and promote cell growth. Topical application can promote wound healing and prevent scar formation and contractures.

Hydrocortisone is a glucocorticoid secreted by the adrenal cortex, which regulates the metabolism of sugar, water, and salt throughout the body. It can be used clinically for anti-inflammation, anti-allergy, and anti-toxin. Hydrocortisone can increase the synthesis of various anti-inflammatory molecular proteins, such as SLPT, and promote the reduction or disappearance of inflammation. In addition, hydrocortisone can also reduce the attack of free radicals on cells by inhibiting the generation of free radicals. Some scholars have found that vasogenic edema is closely related to the scavenging degree of oxygen free radicals in the surrounding extracellular interstitium, and hydrocortisone is a non-enzymatic scavenger of oxygen free radicals, which can directly scavenge free radicals such as OH ₂ and LOO ^- . The hydrocortisone molecular hydrophobic group is embedded in the polyunsaturated fatty acid archway of the membrane bimolecular hydrophobic region, which makes the phospholipid bimolecules tightly combined, increases the viscosity, and blocks the release of arachidonic acid from the cell membrane, thereby reducing the attack of oxygen free radicals. At the same time, hydrocortisone not only inhibits the iron-catalyzed lipid peroxidation reaction in a phospholipid-rich environment, but also inhibits the reaction without the participation of membrane components. Hydrocortisone increases the synthesis and release of lipocertin and macrodermatin through its receptor mediation, inhibits the activity of phosphodiesterase, thereby blocking and inhibiting the prostaglandin cascade reaction, reducing oxygen free radicals and TXA2, white three The formation and release of ene and PG, thereby reducing vascular permeability. Hydrocortisone can inhibit the activity of nitric oxide synthase in macrophages and reduce the production of -NO and oxygen free radicals in macrophages. Combined with other ingredients in biological preparations, it can not only promote the rapid repair of ulcerated tissue, but also protect new tissue from free radical attack and damage.

Human beings suffer from venous diseases such as varicose veins, phlebitis, venous thrombosis, rheumatoid arthritis and diabetes for a long time, resulting in insufficient nutritional supply of the body or local tissues. The later stage of the disease will lead to the symptoms of long-term unhealed skin ulcers/ulceration on the legs and feet to varying degrees, and the prognosis of most of these patients is amputation.

Ulceration/ulceration is the result of local inflammatory necrotic skin decay in the body. Varicose veins/phlebitis ulcers/ulcers are caused by a chronic form of venous insufficiency. Varicose veins of the lower extremities are one of the common surgical diseases, and the prevalence varies greatly from country to country. The incidence rate in many European and American countries is as high as 20%-40%, and it is only 0.1% in tropical Africa. For example, 50% of people over 50 in the UK have mild symptoms of varicose veins, and 10% of them are hospitalized. 5% of adults suffer from chronic venous insufficiency to varying degrees; women have a higher prevalence than men. In China, Zhang Peihua and others surveyed that the incidence rate of people over the age of 15 was 8.6%, and that of people over the age of 45 was 16.4%. Therefore, it is estimated that there are about 80 million to 100 million patients in China. Long-term excessive venous tension or poor venous return will cause venous diseases more often in the femoral vein, saphenous vein and venous part of the leg. 10-25% of patients with varicose veins suffer from arterial disease; 12% of patients with diabetes and rheumatoid arthritis suffer from varicose veins or phlebitis. So far, the cause of varicose veins is unclear, and it may be related to factors such as family history, obesity, pregnancy, and oral contraceptives. At the same time, long-term standing occupations can aggravate the symptoms of varicose veins.

The current methods of treating venous ulcers are as follows: 1. The patient wears two or three (step) highly elastic socks to squeeze the venous blood vessels below the knee to prevent further necrosis; 2. Cross over the affected area Wrap 4 layers of bandages, namely cotton gauze, crepe, Elset and Coban bandages; these two treatments last for about 12 weeks, and about 45% of patients can improve their symptoms, and some patients can still heal their ulcers. 3. Surgical resection: Superficial veins can be removed for patients with superficial varicose veins or phlebitis, but the cure rate is only 2%, and 100% relapse after 4 years. Sometimes the femoral veins that run through the legs can be completely extracted, but The curative effect is not obvious; for patients with deep varicose veins or inflammation, the ulcer is usually removed, the carrion is removed, and the granular skin is transplanted, but it is rarely cured; 4. Laser irradiation treatment for at least 3 months can reduce the pressure of the vein; 5 .Fly maggot therapy, fly maggots can effectively remove wound carrion and help wound healing.

All of the above treatment methods are either cumbersome and time-consuming, or have little effect. Therefore, patients with ulcers or ulcers have to endure pain for many years, causing great pressure to their spirits and bringing great inconvenience to their lives. Moreover, once venous disease causes leg and foot ulceration, if the curative effect of the above treatments is particularly slow or ineffective, the ulceration will continue to expand on the basis of the original necrosis, poor venous return and insufficient nutritional supply will exacerbate this symptom development, eventually leading to high amputation in patients.

Radiation skin ulcers are more common in complications caused by radiation accidents and tumor radiotherapy, which are characterized by intractable recurrent attacks, long-term non-healing, and may eventually develop into cancer. The molecular mechanism of the occurrence and development of this lesion is still unclear, and there is a lack of effective treatment.

Invention content

The object of the present invention is to provide a compound biological preparation of recombinant human epidermal growth factor.

The recombinant human epidermal growth factor compound biological preparation provided by the present invention comprises 2-30ng/ml recombinant human epidermal growth factor, 2-10% D-glucose, 0-20% fetal bovine serum, 0-10 μg/ml insulin, 0 - 5 μg/ml hydrocortisone, 0-100 μg/ml ampicillin and 0-100 IU/ml streptomycin sulfate, the solvent is multiply distilled water.

The amino acid content was calculated based on the added content of fetal bovine serum (FBS, GIBCO).

Wherein, the recombinant human epidermal growth factor compound biological preparation preferably includes 5-10ng/ml recombinant human epidermal growth factor, 5% D-glucose, 0-20% fetal bovine serum, 0-8 μg/ml insulin, 0- 2 μg/ml hydrocortisone, 0-100 μg/ml ampicillin and 0-100 IU/ml streptomycin sulfate.

When the recombinant human epidermal growth factor compound biological preparation is composed of 5ng/ml recombinant human epidermal growth factor, 5% D-glucose, 5 μg/ml insulin, 100IU/ml streptomycin sulfate and multiple distilled water, it can be used for the treatment of ophthalmology disease.

When the recombinant human epidermal growth factor compound biological preparation is composed of 10% fetal bovine serum, 5ng/ml recombinant human epidermal growth factor, 5% D-glucose, 1 μg/ml hydrocortisone, 5 μg/ml or 8 μg/ml insulin , when composed of 100μg/ml ampicillin, 100IU/ml streptomycin sulfate and multiple distilled water, it can be used to treat ulcers caused by varicose veins, phlebitis, etc., and ulcers caused by late complications of diabetes. In order to achieve better therapeutic effect, It can be used with auxiliary treatment solution: 5% D-glucose or 5% D-glucose + 20% FBS, with 20% glycerin aqueous solution + 5% FBS (the latter can be used without FBS).

When the recombinant human epidermal growth factor compound biological preparation is composed of 5% fetal bovine serum, 5ng/ml recombinant human epidermal growth factor, 5% D-glucose, 3 μg/ml insulin, 100 μg/ml ampicillin, 100IU/ml sulfuric acid When composed of streptomycin and multiple distilled water, it can be used to treat healthy wounds, burns and wound healing after surgery. In order to achieve better therapeutic effect, it can be combined with auxiliary treatment solution: 5% D-glucose or 5% D-glucose + 20% FBS use.

When the recombinant human epidermal growth factor compound biological preparation is composed of 5% fetal bovine serum, 5ng/ml recombinant human epidermal growth factor, 5% D-glucose, 3 μg/ml insulin and multiple distilled water, it can resist aging and beautify; In order to achieve a better effect, it can be used with auxiliary liquid: 20% glycerin aqueous solution + 5% FBS.

When the recombinant human epidermal growth factor complex biological preparation is composed of 10ng/ml recombinant human epidermal growth factor, 5% D-glucose, 2 μg/ml hydrocortisone and multiple distilled water, it can be used for treating gastric ulcer.

The recombinant human epidermal growth factor compound biological preparation of the present invention mainly takes recombinant human epidermal growth factor (hrEGF) as the main component, and adds fetal bovine serum (FBS), dextrose (D -glucose), insulin, ampicillin, streptomycin sulfate and hydrocortisone, which not only provide the growth factors required for repair of damaged/healthy tissue, but also provide various nutrients required for rapid tissue repair and promote Cells use the components of these nutrients, so that ulcers or ulcers caused by long-term varicose veins, phlebitis, late diabetes, radiation, etc. can be healed quickly; bedsores and other similar skin wounds can be healed quickly; burns or accidental wounds can be healed quickly; It relieves or even recovers the symptoms of gastric ulcer; makes the rapid recovery of damaging eye diseases; keeps the facial skin tender, reduces wrinkles and dark spots or even disappears. The recombinant human epidermal growth factor compound biological preparation of the present invention can be used to prepare medicines for treating ulcers, ulcers, wounds, and burns; it can also be used to produce anti-aging cosmetics and beauty products to keep skin tender, reduce wrinkles, and darken spots. disappear, and the skin is finally shiny and elastic; it can also be used to prepare medicines for the treatment of damaging eye diseases.

Detailed ways

The following test personnel were all volunteers, and there was no adverse physiological reaction after use; rhEGF was purchased from Sigma Company in the United States, the product number was E 9644, and the content of amino acids was calculated based on the added content of fetal bovine serum (FBS, GIBCO).

Embodiment 1, treatment ophthalmology disease

The formula of the preparation is 5ng/ml rhEGF + 5% D-glucose + 5μg/ml insulin + 100IU/ml streptomycin sulfate, the basic solution is multiple distilled water, after mixing, filter and sterilize, and refrigerate at 4°C.

Before use, it needs to be left at room temperature for about 5 minutes, and then eye drops, one drop at a time, 3-4 times a day. It is especially effective to use it once before going to bed at night. Its effect of treating ophthalmic diseases is shown in Table 1. Before the treatment, the welder's eye burn caused by welding showed as eye congestion, continuous pain, and persisted for a long time. After using this preparation for 1 week, the symptoms disappeared and recovered without rebound.

Table 1. Effects of treating ophthalmic diseases Eye burns caused by welding can't be cured for a long time Year-round bloodshot eyes and dim eyesight caused by unknown reasons number of people 5 people (male) 4 people (male:female=1:1) Effect Symptoms disappeared and healed in 1 week without rebound 1-2 weeks, the symptoms of congestion disappear, and the patient feels good

Embodiment 2, treatment of ulceration caused by varicose veins, phlebitis, etc. and complications of late diabetes:

Main formulation: 10% FBS + 5ng/ml rhEGF + 5% D-glucose + 1μg/ml hydrocortisone + 5μg/ml (8μg/ml) insulin + 100μg/ml ampicillin + 100IU/ml streptomycin sulfate The basic solution is multi-distilled water, which is filtered and sterilized after mixing, and refrigerated at 4°C.

Auxiliary treatment solution: 5% D-glucose + 20% FBS, mixed with 20% glycerin aqueous solution, filtered and sterilized, refrigerated at 4°C.

First use 0.9% normal saline to clean the wound (in severe cases, first use hydrogen peroxide or 75% alcohol to clean), then apply the main preparation to the wound, once a day in the morning and evening. Patients with dry skin or already formed scabs need to use auxiliary treatment solution to keep the wound moist and promote absorption and curative effect. When all patients are using the main preparation, they need to use the auxiliary treatment liquid to smear in the gap between the use of the main preparation to increase the therapeutic effect, 3-4 times a day. For the necrotic tissue surface without ulceration, since the existence of scab hinders the absorption of the active ingredients of the preparation, the main preparation can be injected directly into the active tissue space by subcutaneous injection, once a day. The injection dose should be injected according to the degree of skin absorption, generally 0.05ml-0.1ml; the number of injection points depends on the size of the necrotic area, generally 1 point/cm ² . Table 2 shows the effects of the preparation on treating varicose veins, ulceration caused by phlebitis, ulceration and decubital ulcers caused by late complications of diabetes.

Table 2. The effect of treating varicose veins, ulceration caused by phlebitis, ulceration caused by post-diabetic complications, and bedsores Varicose veins, ulceration caused by phlebitis Ulcers caused by post-diabetic complications bedsore number of people treated 6 people 3 people 1 person treatment effect They are all 60-85-year-olds. After using the main preparation in the morning, the secretion of the ulcerated mouth stops after 4 hours; the granulation tissue of the ulcerated mouth begins to grow three days later. The wound healed in about 2 weeks, and the skin returned to its original color. Insulin in the main preparation was 5 μg/ml. Except that the insulin in the main preparation is 8 μg/ml, the healing method, time and effect are roughly the same as those for treating ulcers caused by varicose veins and phlebitis. The cure method, time and effect are roughly the same as the ulceration caused by treatment of varicose veins and phlebitis.

Embodiment 3, treatment healthy wound, burn and wound healing after operation

Main formulation: 5% FBS + 5ng/ml recombinant human epidermal growth factor + 5% D-glucose + 3μg/ml insulin + 100μg/ml ampicillin + 100IU/ml streptomycin sulfate, the solution was sterilized by filtration, 4°C refrigeration.

Auxiliary treatment solution: 5% D-glucose or 5% D-glucose + 20% FBS.

First use 0.9% normal saline to clean the wound, and then apply the main preparation to the wound, once a day in the morning, twice a day for larger or serious wounds. In order to make the main preparation play a better role, the auxiliary treatment solution is used together to provide sufficient energy and nutrients required for tissue regeneration and repair. Auxiliary treatment solution is used 3-4 times/day. The effects of treating healthy wounds, burns and wound healing after surgery are shown in Table 3.

Table 3. Effects of Treatment on Healthy Wounds, Burns, and Incision Healing After Surgery Burns, burns and trauma Incision after surgery number of people 6 people 3 people treatment effect The recovery of severe burns takes 1.5-3 days; burns can heal 3-5 days earlier than ordinary treatment; trauma generally takes 3-5 days, and the scars are mild. Generally about 3 days, and the scar is slight.

Embodiment 4, anti-aging and beauty treatment

Main formulation: 5% FBS + 5ng/ml recombinant human epidermal growth factor + 5% D-glucose + 3μg/ml insulin, sterilized by filtration, refrigerated at 4°C.

Auxiliary solution: 20% glycerol aqueous solution + 5% FBS, refrigerated at 4°C.

After cleansing the skin every night, use 0.5ml of the main preparation to pat on the face for full absorption. Then, apply an appropriate amount of auxiliary liquid to fully moisturize the skin. The anti-aging and cosmetic effects of the preparation are shown in Table 4.

Table 4. Antiaging and Cosmetic Effects Anti-aging, beauty, anti-freckle and anti-cracking number of people 15 people (adults and children) Effect After 14-20 days of use, the freckles fade or even disappear; the facial skin is delicate, the wrinkles are reduced, and it is shiny; the chappedness disappears, and the skin is as shiny as before.

Embodiment 5, treatment gastric ulcer

Preparation formula: 10ng/ml recombinant human epidermal growth factor + 5% D-glucose + 2 μg/ml hydrocortisone, after autoclaving the glucose aqueous solution, add the other two components when the temperature drops to room temperature.

Take 5-10ml orally on an empty stomach, 2-3 times a day. The effect of the preparation for treating gastric ulcer is shown in Table 5.

Table 5. The effect of treating gastric ulcer gastric ulcer number of people treated 5-people treatment effect Gastroscopy results show that the ulcer surface basically disappears within 1-6 months

Claims (10)

1, a kind of recombinant human epidermal growth factor composite biological agent, comprise the 2-30ng/ml recombinant human epidermal growth factor, the 2-10% D-Glucose, the 0-20% hyclone, 0-10 μ g/ml insulin, 0-5 μ g/ml hydrocortisone, 0-100 μ g/ml ampicillin and 0-100IU/ml streptomycin sulphate, solvent are multiple distilled water.

2, composite biological agent according to claim 1, it is characterized in that: described recombinant human epidermal growth factor composite biological agent, comprise the 5-10ng/ml recombinant human epidermal growth factor, 5% D-Glucose, the 0-20% hyclone, 0-8 μ g/ml insulin, 0-2 μ g/ml hydrocortisone, 0-100 μ g/ml ampicillin and 0-100IU/ml streptomycin sulphate.

3, composite biological agent according to claim 1 and 2, it is characterized in that: described recombinant human epidermal growth factor composite biological agent, by the 5ng/ml recombinant human epidermal growth factor, 5% D-Glucose, 5 μ g/ml insulin, 100IU/ml streptomycin sulphate and aseptic multiple distilled water form.

4, composite biological agent according to claim 1 and 2, it is characterized in that: described recombinant human epidermal growth factor composite biological agent, by 10% hyclone, the 5ng/ml recombinant human epidermal growth factor, 5% D-Glucose, 1 μ g/ml hydrocortisone, 5 μ g/ml or 8 μ g/ml insulin, 100 μ g/ml acillins, 100IU/ml streptomycin sulphate and aseptic multiple distilled water form.

5, composite biological agent according to claim 1 and 2, it is characterized in that: described recombinant human epidermal growth factor composite biological agent, by 5% hyclone, the 5ng/ml recombinant human epidermal growth factor, 5%D-glucose, 3 μ g/ml insulin, 100 μ g/ml acillins, 100IU/ml streptomycin sulphate and aseptic multiple distilled water form.

6, composite biological agent according to claim 1 and 2, it is characterized in that: described recombinant human epidermal growth factor composite biological agent, by 5% hyclone, the 5ng/ml recombinant human epidermal growth factor, 5%D-glucose, 3 μ g/ml insulin and aseptic multiple distilled water form.

7, composite biological agent according to claim 1 and 2, it is characterized in that: described recombinant human epidermal growth factor composite biological agent, by the 10ng/ml recombinant human epidermal growth factor, 5%D-glucose, 2 μ g/ml hydrocortisones and aseptic multiple distilled water form.

8, the described recombinant human epidermal growth factor composite biological agent of claim 1 in preparation treatment because the ulcer that long-term intravenous varicose, phlebitis, diabetes later stage, radioactive ray cause or fester, treatment burn or accident trauma, the application in the medicine for the treatment of bedsore and treatment gastric ulcer.

9, the application of the described recombinant human epidermal growth factor composite biological agent of claim 1 in preparation cosmetics and cosmetics.

10, the application of the described recombinant human epidermal growth factor composite biological agent of claim 1 in the damaging ophthalmology disease of preparation treatment.