CN111228132A

CN111228132A - Skin composite preparation

Info

Publication number: CN111228132A
Application number: CN202010134049.1A
Authority: CN
Inventors: 赵梓纲; 张倩; 严虹; 刘洁; 孔毅
Original assignee: Individual
Current assignee: Individual
Priority date: 2020-03-02
Filing date: 2020-03-02
Publication date: 2020-06-05

Abstract

The invention discloses a skin compound preparation, which comprises the following components: a plurality of cytokines including epidermal growth factor, vascular endothelial growth factor, platelet derived factor, transforming growth factor, fibroblast growth factor, nerve growth factor, and keratinocyte growth factor. In this way, the invention can be applied to skin tissue to slow aging, remove spots and repair damaged skin.

Description

Skin composite preparation

Technical Field

The invention relates to the technical field of medicine and skin care, in particular to a skin composite preparation.

Background

The skin is covered on the surface of human body, and the skin of human body is composed of three layers of epidermis, dermis and subcutaneous tissue. The cells in each layer of the epidermis are tightly connected. The dermis contains a large amount of collagen fibers and elastic fibers, so that the skin is tough and soft and has certain tensile property and elasticity.

However, even then, the skin of the human body is often exposed to various problems due to various intrinsic and extrinsic factors, including extrinsic trauma, injury, intrinsic inflammation, immune response, etc., resulting in various problems of aging, damage, dark spots, wrinkles, etc., of the skin tissue.

Disclosure of Invention

The invention mainly solves the technical problem of providing a skin composite preparation which can be used for skin tissues to slow down aging, remove freckles and repair damaged skin.

In order to solve the technical problems, the invention adopts a technical scheme that: provided is a skin complex preparation comprising: a plurality of cytokines including epidermal growth factor, vascular endothelial growth factor, platelet derived factor, transforming growth factor, fibroblast growth factor, nerve growth factor, and keratinocyte growth factor.

Wherein the skin complex formulation further comprises water.

Wherein the skin complex formulation further comprises a soluble salt.

Wherein the salts comprise sodium salt, potassium salt and zinc salt; the salts comprise sodium salts and zinc salts.

Wherein the zinc salt comprises zinc chloride or a hydrated salt thereof, zinc sulfate or a hydrated salt thereof, and zinc nitrate or a hydrated salt thereof; the sodium salt includes sodium chloride and heparin sodium.

Wherein the mass ratio of the epidermal growth factor, the vascular endothelial growth factor, the platelet derived factor, the transforming growth factor, the fibroblast growth factor, the nerve growth factor, the keratinocyte growth factor, the water and the salt is 1-5: 60-80: 1-10; the mass ratio is 3: 75: 7.

Wherein, the skin compound preparation acts on the skin by means of external application or superficial injection.

In order to solve the technical problem, the invention adopts another technical scheme that: there is provided a composite preparation for treating or preventing atopic dermatitis, comprising an anti-atopic dermatitis effective amount of the skin composite preparation as described in any one of the above and a pharmaceutically acceptable carrier.

Wherein the complex formulation is in dosage unit form.

The invention has the beneficial effects that: in contrast to the prior art, the skin complex formulation of the present invention includes various cytokines, and the epidermal growth factor can promote the proliferation and differentiation of cells, thereby substituting the senescent and dead cells with the novacells; the vascular endothelial growth factor has the functions of promoting the increase of vascular permeability, the degeneration of extracellular matrix, the migration and proliferation of vascular endothelial cells, the formation of blood vessels and the like; the platelet-derived factor has the capacity of stimulating the division and proliferation of specific cell groups and promoting the generation of fibroblasts, thereby repairing the breakage and deformation of collagen fibers in the dermis layer caused by aging and damage, promoting the growth and the elasticity improvement of the dermis layer and ensuring that wrinkles naturally grow flat; the transforming growth factor can promote the expression of extracellular matrix (ECM) such as collagen and fibronectin and inhibit the degradation of ECM, plays an important role in the morphogenesis, proliferation and differentiation processes of cells and is beneficial to cell repair; fibroblast growth factor contributes to the synthesis of elastic fibers, reticular fibers and collagen; the nerve growth factor can convert nutrition and inhibit hormone dependence; keratinocyte growth factor can specifically stimulate physiological processes of metabolism of epithelial cells, including regeneration, differentiation, migration and the like of the cells. Through the mode, the skin composite preparation can be used for skin tissues to relieve aging and remove freckles and repair damaged skin.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts. Wherein:

FIG. 1 is a schematic diagram showing comparison photographs before and after use of 3 freckle-removing patients selected in an experiment for freckle-removing effect on skin;

FIG. 2 is a photograph showing comparison of pre-use and post-use photographs of 1 patient with sensitive facial skin selected in an effect test for repairing sensitive skin;

FIG. 3 is a schematic diagram of a cell experiment in an experiment for the effect of accelerating wound healing;

fig. 4 is a photograph showing comparison of pre-and post-use photographs of 2 patients with facial acne selected in an experiment for effect of accelerating wound healing.

Detailed Description

The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

The invention provides a skin composite preparation, which comprises the following components: a plurality of cytokines including epidermal growth factor, vascular endothelial growth factor, platelet derived factor, transforming growth factor, fibroblast growth factor, nerve growth factor, and keratinocyte growth factor.

Epidermal Growth Factor (EGF), also known as human oligopeptide-1, is an active substance in human body, and an active polypeptide consisting of 53 amino groups can repair and proliferate Epidermal cells by stimulating tyrosine phosphorylation of Epidermal Growth Factor receptor. The main characteristic is that it can promote the proliferation and differentiation of cells, so that the new cells can replace the aged and dead cells. EGF also stops bleeding, and has effects of accelerating wound healing of skin and mucosa, relieving inflammation and pain, and preventing ulcer. The EGF has excellent stability, is not easy to lose and flow at normal temperature, and can form good coordination effect with various enzymes in human bodies. EGF can promote the growth and division of various cells in epidermal cell tissues, so that epidermal cells become full and recover a young state, and can also promote the growth capacity of collagen and repair aged and broken collagen elastic fibers. At a young age, the human body can produce enough EGF to promote the growth of epidermal cells, but with the age, the production of EGF is gradually reduced, so that the renewal speed of epidermal cells is slower and slower. Since EGF is a component of human body, it has no toxic side effect and is very safe.

Vascular Endothelial Growth Factor (VEGF), also known as Vascular Permeability Factor (VPF), is a highly specific Vascular Endothelial cell Growth Factor that has the effects of promoting increased Vascular permeability, extracellular matrix degeneration, Vascular Endothelial cell migration, proliferation, and angiogenesis. VEGF is a specific mitogen for vascular endothelial cells, and can promote the growth of vascular endothelial cells in vitro and induce vascular proliferation in vivo. Especially under the hypoxia environment, VEGF is combined with VEGF receptor on endothelial cell membrane to cause the autophosphorylation of the receptor, thereby activating Mitogen-Activated protein kinase (MAPK), realizing the Mitogen property of VEGF and inducing the proliferation of endothelial cells.

The PDGF is a new skin-wrinkle-removing anti-aging new fibroblast acting on a dermis layer and can be really absorbed by receptor cells through dermis layer micro-intervention to achieve a good anti-aging and wrinkle-removing effect, the PDGF is an important mitogenic factor which has the capacity of stimulating the proliferation of a specific cell group, promotes the generation of fibroblasts, repairs the fibroblast, a damaged dermis layer, delays the growth of a fibroblast, and delays the growth of a collagen, and the PDGF is an important skin-stimulating factor which has the functions of delaying the growth of a collagen, promoting the skin aging, and enhancing the skin aging, and the skin aging.

TGF- α and TGF- β, TGF- α is produced by macrophage, brain cell and epidermal cell, and can induce epithelial development, human TGF- β has three subtypes, TGF- β 1, TGF- β 2 and TGF- β, TGF- β is a multifunctional protein, and can affect the Growth, differentiation, apoptosis, immunoregulation and other functions of various cells, TGF- β has potential application prospect in the aspects of treating wound healing, promoting cartilage and bone repair, treating autoimmune diseases through immunosuppression, transplant rejection and the like.

Fibroblast Growth Factor (FGF), a Fibroblast Growth Factor (FGF), is capable of promoting neovascularization and repairing damaged endothelial cells. Has the function of deep repair, and plays an immeasurable great role in modern clinical medicine, surgical operation and cosmetic operation.

Nerve Growth Factor (NGF), a class of protein molecules produced by tissues innervated by nerves (e.g., muscle) and astrocytes and essential for the Growth and survival of neurons.

The Keratinocyte Growth Factor (KGF) is a basic protein Growth Factor secreted by human subcutaneous tissue cells, can specifically stimulate the physiological processes of metabolism of epithelial cells and the like, including regeneration, differentiation, migration and the like of the cells, the KGF has the advantages of strong tissue specificity, stability and good safety of ①, the KGF only acts on the epithelial cells, can start the coordinated signal feedback of the epithelial cells to subcutaneous interstitial tissues while stimulating the Growth of the epithelial cells through a specific receptor, promotes the formation of new tissues, has good safety and no potential side effect, ② has the effect of stimulating the Growth and division of the new or aged epithelial cells, ③ has the obvious effects of removing scars and resisting radiation damage, and ④ has unique hair growing and growing effects.

The skin compound preparation comprises a plurality of cytokines, and the epidermal growth factor can promote the proliferation and differentiation of cells so as to replace aged and dead cells with new cells; the vascular endothelial growth factor has the functions of promoting the increase of vascular permeability, the degeneration of extracellular matrix, the migration and proliferation of vascular endothelial cells, the formation of blood vessels and the like; the platelet-derived factor has the capacity of stimulating the division and proliferation of specific cell groups and promoting the generation of fibroblasts, thereby repairing the breakage and deformation of collagen fibers in the dermis layer caused by aging and damage, promoting the growth and the elasticity improvement of the dermis layer and ensuring that wrinkles naturally grow flat; the transforming growth factor can promote the expression of extracellular matrix (ECM) such as collagen and fibronectin and inhibit the degradation of ECM, plays an important role in the morphogenesis, proliferation and differentiation processes of cells and is beneficial to cell repair; fibroblast growth factor contributes to the synthesis of elastic fibers, reticular fibers and collagen; the nerve growth factor can convert nutrition and inhibit hormone dependence; keratinocyte growth factor can specifically stimulate physiological processes of metabolism of epithelial cells, including regeneration, differentiation, migration and the like of the cells. Through the mode, the skin composite preparation can be used for skin tissues to relieve aging and remove freckles and repair damaged skin.

Wherein the skin complex formulation further comprises water. On the one hand, the various cytokines are cheaper, easier to obtain and safer than other solvents, and on the other hand, the various cytokines are more easily absorbed by the skin when dissolved in water. In addition, the mixture of various cytokines can be stored and transported in the form of freeze-dried powder so as to ensure the effective period of the mixture of various cytokines, and the mixture is simple and convenient to redissolve with water.

In one embodiment, the skin complex formulation further comprises soluble salts, which are capable of shrinking pores while maintaining some protein molecules stable, balancing the osmotic pressure of the solution, and allowing the ingredients of the skin complex formulation to better penetrate into the skin. Further, the salts include sodium salt, potassium salt, zinc salt; the salts include sodium salt and zinc salt.

Wherein the zinc salt comprises zinc chloride or hydrated salt thereof, zinc sulfate or hydrated salt thereof, and zinc nitrate or hydrated salt thereof; the sodium salt includes sodium chloride and heparin sodium.

In one embodiment, the mass ratio of the epidermal growth factor, the vascular endothelial growth factor, the platelet-derived factor, the transforming growth factor, the fibroblast growth factor, the nerve growth factor, the keratinocyte growth factor, the water and the salt is 1-5: 60-80: 1-10; for example, 1: 60: 1, 3: 70: 5, 5: 80: 10, and the like. In one application, the mass ratio is 3: 75: 7.

The skin compound preparation can be used as health product or cosmetic, and can be added with necessary adjuvants, excipient, flavoring agent or their combination. When the skin composite preparation acts on skin, the skin composite preparation can slow down skin tissue aging, repair damaged skin, remove melanocytes and remove freckles. Usually, the skin complex preparation is applied to the skin by means of external application or superficial injection. The action mode is simple and convenient and is convenient to implement.

The application also provides a composite preparation for treating or preventing allergic dermatitis, which comprises the skin composite preparation of any one of the above items and a pharmaceutically acceptable carrier, wherein the skin composite preparation has an effective dose for resisting the allergic dermatitis.

Allergic dermatitis is a skin disease caused by allergens, and mainly refers to skin diseases such as red and swollen skin, itching, wheezy and desquamation and the like caused by contacting human bodies with certain allergens.

Wherein the complex formulation is in dosage unit form.

For the above therapeutic uses, the dosage administered will, of course, vary with the mode of administration, the treatment desired, and the disease condition indicated. The total daily dose may be administered in a single dose or in divided doses.

The compound preparation for treating or preventing allergic dermatitis can be used as a medicine product, and necessary auxiliary materials, excipients, spices or combination thereof can be added when the medicine product is specifically prepared. When the pharmaceutical product is applied on skin, it can slow down skin tissue aging, repair damaged skin, remove melanocyte, and remove speckle. The pharmaceutical product is typically applied to the skin by topical application or by superficial injection. The action mode is simple and convenient and is convenient to implement.

The various cytokines can be obtained by adopting a stem cell culture mode, and then the skin composite preparation is prepared.

Specifically, the method comprises the following steps:

step (1): culturing stem cells (according to the culture condition of normal cells, 37 ℃ and 5% carbon dioxide) for 24 hours, and taking supernatant; detecting the concentration of each cytokine by ELISA;

step (2): culturing the stem cells for 36 hours, and taking supernatant; detecting the concentration of each cytokine by ELISA;

and (3): culturing the stem cells for 48 hours, and taking supernatant; detecting the concentration of each cytokine by ELISA;

and (4): culturing the stem cells for 72 hours, and taking supernatant; detecting the concentration of each cytokine by ELISA;

and (5): centrifuging stem cell culture solution at different time to collect protein components; dissolving in matched water; in the stem cell culture, the seven cytokines are the components with the largest content in the stem cell culture solution, and the concentrations of the seven cytokines obtained by the stem cell culture at the same time are different, so that the effective proportions of various cytokines can be obtained by culturing the stem cells at different times.

And (6): adjusting the pH value of the system to 6.0-7.5 by using a pH buffer solution; the pH buffer may be: conventional buffer solutions such as sodium citrate buffer solution, sodium bicarbonate buffer solution, PBS buffer solution, and the like; further, the pH buffer is sodium bicarbonate buffer.

The specific effects of the composite preparation of the present embodiment for removing freckles, repairing sensitive skin, and healing wounds will be described in detail below.

Example 1 Effect test for removing skin spots

Volunteer grouping conditions: selecting a facial freckle patient;

the number of volunteers and the specific case of the volunteers were selected from 100 volunteers, 50 male and female, and the age was 20-50 years.

The product is as follows: the skin composite preparation is prepared into a facial mask form, wherein the mass ratio of epidermal growth factor, vascular endothelial growth factor, platelet derived factor, transforming growth factor, fibroblast growth factor, nerve growth factor, keratinocyte growth factor, water and salt is 3: 75: 7.

The use method comprises the following steps: after the face is cleaned every night, the face is applied to the face for 20 minutes for 14 days continuously, and freckles can be obviously reduced. The specific method comprises the following steps: the freckle area is measured by adopting a number grid method, and specifically, a number counting method of printing a color smearing paper and a millimeter check coordinate paper is adopted. Tracing the outline of the skin damage to be measured by using a marker pen, then laying a drawing with a standard coordinate grid on the drawing, counting the number of the grids by visual observation (counting by one grid when the number of the grids is less than one grid), multiplying the area of each small grid to obtain the area of the skin damage, repeatedly measuring for 3 times, and taking the average value as the final freckle area measurement value. Referring to fig. 1, fig. 1 is a schematic diagram showing a comparison between before and after use of 3 spot-removing patients selected in an experiment for removing spots on skin. As is apparent from FIG. 1, the number of freckles was significantly reduced after the administration of 3 patients.

Example 2 Effect test for repairing sensitive skin

Volunteer grouping conditions: selecting a patient with sensitive facial skin;

the number of volunteers and the specific conditions were as follows: selecting 50 volunteers, 25 male and female, and 20-50 years old.

The use method comprises the following steps: after the face is cleaned at night every day, the face-washing cream is applied to the face for 20 minutes for 14 days continuously, the symptoms of sensitive skin can be relieved, the redness of the face is obviously improved, the skin is not easy to be reddened before, the skin is not easy to be reddened under common temperature change, supercooling or overheating, and the skin is not easy to be reddened and generate heat. Referring to fig. 2, fig. 2 is a photograph illustrating comparison between before and after use of 1 patient with sensitive facial skin selected in the effect test for repairing sensitive skin. As is evident from FIG. 2, after the patient took the medicine, the redness of the face was significantly improved, the area of the face was reduced, and the symptoms were alleviated.

Example 3, effect experiments for accelerating wound healing, including cell experiments and acne experiments:

cell experiments:

(1) epidermal cell line A-431 was cultured in 5% CO₂Culturing at 37 ℃;

(2) after the cells adhered to the wall, a scratch experiment was performed, and the cells were divided into two groups, one group was a blank control group (medium is only incomplete medium), and the other group was a cytokine group (incomplete medium + skin complex preparation), and the healing speed of the cells was observed: 0h, 6h, 12h, 24h, 36h and 48 h. The results of the experiment are shown in FIG. 3. The top row in fig. 3 is a blank group and the bottom group is a combined formulation group, as is evident from fig. 3: the healing of the compound preparation group is very obvious from 0h to 48 h; while the blank group had a slow healing rate from 0h to 48 h.

Acne experiments:

volunteer grouping conditions: : selecting a patient with facial acne;

the number of volunteers and the specific conditions were as follows: 100 volunteers are selected, and the male and the female are 50 people and 20-50 years old respectively.

The use method comprises the following steps: after the face was cleaned every night and applied to the face for 20 minutes for 14 consecutive days, it was seen that the rash was remarkably subsided and the subjective red swelling and thermal pain of the patient was relieved, see fig. 4, which is a photograph showing 2 patients with facial acne before and after use compared before and after use selected in the experiment for the effect of accelerating wound healing. As is evident from fig. 4, the 2 patients had significant regression of the facial sores after use.

The above description is only a preferred embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by using the contents of the present specification and the accompanying drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.

Claims

1. A skin complex formulation comprising: a plurality of cytokines including epidermal growth factor, vascular endothelial growth factor, platelet derived factor, transforming growth factor, fibroblast growth factor, nerve growth factor, and keratinocyte growth factor.

2. The skin complex formulation of claim 1, further comprising water.

3. The skin complex formulation of claim 2, further comprising a soluble salt.

4. The skin complex formulation of claim 3, wherein the salts comprise sodium, potassium, zinc salts; the salts comprise sodium salts and zinc salts.

5. The skin complex formulation according to claim 4, wherein the zinc salt comprises zinc chloride or a hydrated salt thereof, zinc sulfate or a hydrated salt thereof, zinc nitrate or a hydrated salt thereof; the sodium salt includes sodium chloride and heparin sodium.

6. The skin composite preparation according to claim 3, wherein the mass ratio of the epidermal growth factor, the vascular endothelial growth factor, the platelet-derived factor, the transforming growth factor, the fibroblast growth factor, the nerve growth factor, the keratinocyte growth factor, the water and the salts is 1-5: 60-80: 1-10; the mass ratio is 3: 75: 7.

7. The skin complex formulation according to any one of claims 1 to 6, wherein the skin complex formulation is applied to the skin by means of external application or superficial injection.

8. A composite preparation for treating or preventing atopic dermatitis, which comprises the skin composite preparation according to any one of claims 1 to 7 in an amount effective for the treatment of atopic dermatitis and a pharmaceutically acceptable carrier.

9. The combination formulation of claim 8, wherein the combination formulation is in dosage unit form.