CN109045279B

CN109045279B - External composition, application thereof and external therapeutic agent

Info

Publication number: CN109045279B
Application number: CN201811054306.XA
Authority: CN
Inventors: 李乾
Original assignee: Cause Color Biotechnology (wuhan) Co Ltd
Current assignee: Jiangsu Yuan Pharmaceutical Research Co.,Ltd.
Priority date: 2018-09-10
Filing date: 2018-09-10
Publication date: 2020-01-21
Anticipated expiration: 2038-09-10
Also published as: CN109045279A

Abstract

The invention discloses an external composition, application thereof and an external therapeutic agent. The external composition is prepared from the recombinant human metallothionein and the resveratrol, and when the external composition is applied to an external therapeutic agent for treating infantile eczema and/or diaper dermatitis, the external composition can effectively relieve skin inflammation and adjust skin immune function, and has a remarkable treatment effect on the infantile eczema and the diaper dermatitis through the combination of the recombinant human metallothionein and the resveratrol.

Description

External composition, application thereof and external therapeutic agent

Technical Field

The invention relates to the technical field of biological medicines, and particularly relates to an external composition, application thereof and an external therapeutic agent.

Background

The skin of infants is delicate, the infants often suffer from infantile eczema shortly after birth without reason, the infants can suffer from the whole body, the symptoms of red papules, itching, easy liquid seepage and scabbing after scratching, and particularly the scalp often forms a large gray thick scab which is not easy to fall off. On the other hand, even when an infant or a young child uses a diaper for a long period of time, diaper dermatitis, which is manifested as redness, or rash, is likely to occur due to the friction between the diaper and the skin irritation caused by urine dipping. However, no product which is quick and effective for treating infantile eczema and diaper dermatitis exists in the existing medicines in the market at present.

Disclosure of Invention

The invention mainly aims to provide an external composition, application thereof and an external therapeutic agent, and aims to provide an external therapeutic agent capable of quickly and effectively treating infantile eczema and diaper dermatitis.

In order to achieve the purpose, the invention provides an external composition which comprises recombinant human metallothionein and resveratrol.

Preferably, the recombinant human metallothionein is any one of recombinant human metallothionein-1 and a subtype and a mutant thereof as well as a fusion protein of the recombinant human metallothionein-1; or the like, or, alternatively,

the recombinant human metallothionein is any one of recombinant human metallothionein-2 and subtype and mutant thereof as well as fusion protein of the recombinant human metallothionein-2; or the like, or, alternatively,

the recombinant human metallothionein is any one of recombinant human metallothionein-3 and subtype and mutant thereof as well as fusion protein of the recombinant human metallothionein-3; or the like, or, alternatively,

the recombinant human metallothionein is any one of recombinant human metallothionein-4 and subtype and mutant thereof, and fusion protein of the recombinant human metallothionein-4.

The invention also provides the application of the external composition in preparing an external therapeutic agent for treating infantile eczema and/or diaper dermatitis.

The invention also provides an external therapeutic agent, which comprises the external composition.

Preferably, the addition amount of the recombinant human metallothionein in the external therapeutic agent is 0.01-0.06 g/kg;

the addition amount of the resveratrol in the external therapeutic agent is 1-10 g/kg.

Preferably, the addition amount of the recombinant human metallothionein in the external therapeutic agent is 0.02-0.05 g/kg;

the addition amount of the resveratrol in the external therapeutic agent is 3-7 g/kg.

Preferably, the addition amount of the recombinant human metallothionein in the external therapeutic agent is 0.025-0.04 g/kg;

the addition amount of the resveratrol in the external therapeutic agent is 5-6 g/kg.

Preferably, the external therapeutic agent is a gel, an emulsion, a cream, a solution, a liniment, a lotion, an ointment, or an ointment.

According to the technical scheme provided by the invention, the external composition is prepared from the recombinant human metallothionein and resveratrol, wherein the recombinant human metallothionein can combine and detoxify harmful heavy metals, the adaptability of cells to external adverse factor stimulation is regulated by neutralizing free radicals, inflammatory reaction is relieved, skin repair is facilitated, other proteins or compounds are protected from being oxidized and inactivated too fast, resveratrol has the effects of resisting oxidation, resisting free radicals, resisting bacteria and regulating an immune system, and when the external composition is applied to an external therapeutic agent for treating infantile eczema and/or diaper dermatitis, the skin inflammation and the skin immune function can be effectively relieved, and the therapeutic effect on the infantile eczema and the diaper dermatitis is remarkable and quick.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

The invention provides an external composition, which comprises recombinant human metallothionein and resveratrol.

The main application of the external composition is to prepare an external therapeutic agent for treating infantile eczema and/or diaper dermatitis. It is understood that the external composition may be formulated with pharmaceutically acceptable salts or carriers thereof and then mixed with conventional external skin preparations (e.g., emulsion preparations, cream preparations, etc.) to prepare an external therapeutic agent, or may be directly mixed with conventional external skin preparations to prepare an external therapeutic agent, when the external composition is applied to an external therapeutic agent for treating infantile eczema and/or diaper dermatitis.

In the composition for external use, the functions of the components are as follows:

the Recombinant Human Metallothionein (rhMT) is derived from Human Metallothionein expressed by genetic engineering recombination, and the rhMT is homologous with the amino acid sequence of Human self MT, so that the defect that the allergy is caused by foreign proteins is avoided. The rhMT can combine and detoxify harmful heavy metals, can regulate the adaptability of cells to external harmful factor stimulation by neutralizing free radicals, relieves inflammatory reaction, is favorable for promoting epidermal repair and is favorable for protecting other proteins or compounds from being oxidized and inactivated too fast.

Resveratrol is a plant-derived polyphenolic compound, also known as Chinese: 3,4', 5-trihydroxystilbene; tiger stick aglycone; a stilbenetriol; resveratrol; 3,4', 5-trihydroxy stilbene; 3,4', 5-trihydroxy stilbene; (E) -5- [2- (4-hydroxyphenyl) -vinyl ] -1, 3-benzenediol, 3,4, 5-trihydroxy trans (E) -3,5,4' -trihydroxystilbene. Resveratrol has the following relevant biological activities: (1) resveratrol is a natural antioxidant existing in plants, and plays an antioxidant action mechanism mainly including the functions of eliminating or inhibiting the generation of free radicals, inhibiting lipid peroxidation, regulating the activity of antioxidant-related enzymes and the like. In recent years, attention has been paid to physiological functions of resveratrol for resisting oxidation and scavenging free radicals, because the physiological metabolism involves many diseases such as atherosclerosis, senile dementia, viral hepatitis and the like which are closely related to human health, and therefore, resveratrol is known to have the effects of resisting oxidation and free radicals. (2) Resveratrol is known as a natural phytoalexin, and when plants such as grapes are subjected to adverse conditions such as fungal infection and ultraviolet irradiation, resveratrol in the corresponding parts is accumulated to cope with the adverse conditions, and thus resveratrol is known to have an antibacterial effect. (3) Researches find that resveratrol with different doses can obviously improve macrophage phagocytosis rate, serum half-hemolysis value, organism antibody forming cell number, lymphocyte transformation rate and inhibition of mouse ear swelling degree (namely IV type hypersensitivity of normal mice) of an immunosuppression model mouse, and for the normal mice, resveratrol has no obvious influence on immune function but can inhibit IV type hypersensitivity, so that the resveratrol has a regulating effect on an immune system.

In the technical scheme provided by the invention, rhMT is used as a component in the external composition and is derived from a human body, but not derived from MT protein extracted from other animals, plants or microorganisms, so that the possible damage of immunogenicity to skin caused by amino acid sequence difference of heterologous protein is avoided. Meanwhile, the rhMT can be combined with and detoxify harmful heavy metals, the adaptability of cells to external harmful factor stimulation is regulated by neutralizing free radicals, the inflammatory reaction is relieved, the skin repair is facilitated, the characteristics of excessive oxidation inactivation of other proteins or compounds are protected, the rhMT is compatible with resveratrol, the antioxidation effect of the rhMT is further enhanced by utilizing the effects of resveratrol on antioxidation, anti-free radical, antibiosis and immune system regulation, and the external composition formed by combining the rhMT and the resveratrol can effectively relieve the skin inflammation and regulate the skin immune function when being applied to an external therapeutic agent for treating infantile eczema and/or diaper dermatitis, and has obvious and quick treatment effect on the infantile eczema and the diaper dermatitis.

In an embodiment of the external composition provided by the present invention, the rhMT is any one of rhMT-1 and its subtype, mutant and fusion protein of rhMT-1; or the rhMT is any one of rhMT-2 and subtype and mutant thereof as well as fusion protein of the rhMT-2; or the rhMT is any one of rhMT-3 and subtype and mutant thereof as well as fusion protein of the rhMT-3; or the rhMT is any one of rhMT-4 and subtype and mutant thereof and fusion protein of the rhMT-4. Any one of the rhMT, the subtype and the mutant thereof and the fusion protein of the rhMT is selected as the rhMT to be compatible with the resveratrol, so that the external composition with remarkable treatment effect on the infantile eczema and/or the diaper dermatitis can be prepared.

The following description will be given taking as an example that the external composition is directly mixed with a conventional external skin preparation to prepare an external therapeutic agent for treating infantile eczema and/or diaper dermatitis when used for preparing the external therapeutic agent. The invention also provides an external therapeutic agent, which comprises the external composition. In an embodiment of the external therapeutic agent provided by the present invention, the external therapeutic agent further includes a material body, the material body is preferably a material body that is soft and has no irritation to delicate skin of infants, any material body that is conventionally used for skin external agents in the field may be selected, or the external therapeutic agent may be self-prepared according to general components of existing skin external therapeutic agent bodies, the components of the external therapeutic agent generally include water, oil, grease, moisturizers, surfactants, thickeners, skin conditioners, pH regulators, preservatives, and the like, and the selection or the preparation of the material body is prior art in the field and will not be described herein.

In a preferred embodiment of the external therapeutic agent provided by the present invention, the addition amount of the rhMT in the external therapeutic agent is 0.01 to 0.06g/kg, and the addition amount of the resveratrol in the external therapeutic agent is 1 to 10g/kg, which is: each kilogram of the external therapeutic agent contains 0.01-0.06 g of rhMT and 1-10 g of resveratrol. The external therapeutic agent prepared from the rhMT and the resveratrol within the addition amount range can effectively relieve or eliminate infant skin discomfort caused by infant eczema and/or diaper dermatitis after being coated on skin, and has the advantages of quick response and no adverse reaction.

In another preferred embodiment of the external therapeutic agent provided by the present invention, the addition amount of the rhMT in the external therapeutic agent is 0.02 to 0.05g/kg, and the addition amount of the resveratrol in the external therapeutic agent is 3 to 7g/kg, which is: the rhMT and resveratrol content in each kilogram of the external therapeutic agent is 0.02-0.05 g and 3-7 g respectively. The external therapeutic agent prepared from the rhMT and the resveratrol within the addition range has better treatment effect on infantile eczema and/or diaper dermatitis.

In another preferred embodiment of the external therapeutic agent provided by the present invention, the addition amount of the rhMT in the external therapeutic agent is 0.025 to 0.04g/kg, and the addition amount of the resveratrol in the external therapeutic agent is 5 to 6g/kg, which is: the rhMT and resveratrol content of each kilogram of the external therapeutic agent is 0.025-0.04 g and 5-6 g respectively. The external therapeutic agent prepared by the rhMT and the resveratrol within the adding amount range has excellent treatment effect on infantile eczema and/or diaper dermatitis.

During the preparation process of the external therapeutic agent, the external composition can be added into a material body in the forms of gel, emulsion, cream, solution, paste and the like to be uniformly mixed, and the mixture is correspondingly prepared into gel, emulsion, cream, solution, liniment, lotion, ointment or paste so as to meet the use requirements of different users. The dosage form of the external therapeutic agent is not limited to the above dosage forms, but the external composition is added, so that the external therapeutic agent is convenient for daily use of a user, has a remarkable curative effect on infantile eczema and/or diaper dermatitis, and has no toxic or side effect.

The technical solutions of the present invention are further described in detail with reference to the following specific examples, which should be understood as merely illustrative and not limitative.

EXAMPLE 1 preparation of external ointment

(1) Weighing raw materials: 0.04g of recombinant human metallothionein freeze-dried powder, 6g of resveratrol and 1kg of ointment body, wherein the formula of the ointment body is shown in the following table 1.

TABLE 1 topical ointment body formulations

(2) Preparation of external ointment: adding the raw materials for preparing the ointment body into a heating stirrer according to the formula in the table 1, heating to 80 ℃, stirring at the rotating speed of 100rpm for 10min, then cooling to 50 ℃, adding resveratrol into the stirrer, continuously stirring at the rotating speed of 80rpm for 10min, then cooling again, after the temperature of the ointment body is reduced to below 40 ℃, adding the recombinant human metallothionein freeze-dried powder into the stirrer, stirring at the rotating speed of 40rpm for 5min, preparing the external ointment, discharging and packaging.

Example 2 preparation of external cream

(1) Weighing raw materials: 0.03g of recombinant human metallothionein freeze-dried powder, 6g of resveratrol and 1kg of cream-like material body, wherein the formula of the cream-like material body is shown in the following table 2.

TABLE 2 external cream body formula

(2) Preparation of external cream: adding the raw materials for preparing the cream-like material body into a heating stirrer according to the formula in the table 2, heating to 80 ℃, stirring at the rotating speed of 100rpm for 10min, then cooling to 50 ℃, adding resveratrol into the stirrer, continuously stirring at the rotating speed of 80rpm for 10min, then cooling again, after the temperature is reduced to below 40 ℃, adding the recombinant human metallothionein freeze-dried powder into the stirrer, stirring at the rotating speed of 40rpm for 5min, preparing the external ointment, discharging and packaging.

EXAMPLE 3 preparation of external emulsion

(1) Weighing raw materials: 0.025g of recombinant human metallothionein freeze-dried powder, 5g of resveratrol and 1kg of emulsion body, wherein the formula of the emulsion body is shown in the following table 3.

TABLE 3 topical emulsion body formulation

(2) Preparation of external emulsion: adding the raw materials for preparing the emulsion body into a heating stirrer according to the formula in the table 3, heating to 80 ℃, stirring at the rotating speed of 100rpm for 10min, then cooling to 50 ℃, adding resveratrol into the stirrer, continuously stirring at the rotating speed of 80rpm for 10min, then cooling again, after the temperature is reduced to below 40 ℃, adding the recombinant human metallothionein freeze-dried powder into the stirrer, stirring at the rotating speed of 40rpm for 5min, preparing the external ointment, discharging and packaging.

Example 4 preparation of external nanoemulsion essence

(1) Weighing raw materials: 0.02g of recombinant human metallothionein freeze-dried powder, 5g of resveratrol and 1kg of essence body, wherein the formula of the essence body is shown in the following table 4.

Table 4 formula of external essence body

(2) Preparing external nano-emulsion essence:

a. adding allantoin (No. 10 raw material in Table 4) into a heat-resistant microwave-resistant plastic bottle, adding 150mL of water, heating to 80-90 ℃, covering a bottle, covering the bottle, shaking until the powder is completely dissolved, and preparing an allantoin solution for later use;

b. equally dividing glycerol (raw material No. 2 in table 4) into two bottles, adding sodium hyaluronate (raw material No. 8 in table 4) into one of the two bottles, and fully and uniformly mixing to prepare a glycerol-sodium hyaluronate mixture for later use; mixing another bottle of glycerol with hydrogenated olive oil, tocopherol acetate, bisabolol and tween-80 (No. 4,5, 6 and 7 raw materials in Table 4), adding resveratrol, shaking and mixing for 2min, pouring into water, emulsifying at high speed of 5000rpm for 5min, standing for 10min for defoaming, and making into mixed solution;

c. mixing the obtained allantoin solution, glycerol-sodium hyaluronate mixture and mixed solution with all the rest raw materials in Table 4, stirring at 100rpm for 10min, adding recombinant human metallothionein lyophilized powder, stirring at 80rpm for 5min to obtain topical nanoemulsion essence, discharging, and packaging. The prepared nano-emulsion essence is clear and transparent, and has no phenomenon of oil water stratification or oil floating after standing for 7 days at room temperature.

Example 5

The same as example 1, except that the mass of the recombinant human metallothionein freeze-dried powder and the mass of the resveratrol which are weighed in the step (1) are respectively 0.035g and 5.5 g.

Example 6

The same as example 1, except that the mass of the recombinant human metallothionein freeze-dried powder and the mass of the resveratrol which are weighed in the step (1) are respectively 0.05g and 3 g.

Example 7

The same as example 1, except that the mass of the recombinant human metallothionein freeze-dried powder and the mass of the resveratrol weighed in the step (1) are respectively 0.05g and 7 g.

Example 8

The same as example 1, except that the mass of the recombinant human metallothionein freeze-dried powder and the mass of the resveratrol which are weighed in the step (1) are respectively 0.01g and 1 g.

Example 9

The same as example 1, except that the mass of the recombinant human metallothionein freeze-dried powder and the mass of the resveratrol weighed in the step (1) are respectively 0.06g and 10 g.

The external ointment prepared in the above example 1 is taken as an example to verify the efficacy of the external therapeutic agent provided by the present invention in treating infantile eczema and diaper dermatitis, and the test method and results are as follows:

1. therapeutic effect on infantile eczema

18 clinical follow-up patients with infantile eczema, the age of 3-12 months, the external essence prepared in the embodiment 4 is applied to affected parts (eczema is preferred), the external essence is applied to the affected parts once a day in the morning and at night, after the external essence is used for 3 days, the skin congestion of all children patients is relieved, after the external essence is used for 5 days, the eczema of all children patients disappears, and no recurrence is seen in follow-up visit within 1 year. The external essence provided by the embodiment of the invention has the effective rate of 100% for treating infantile eczema, and is quick in effect, skin congestion of a patient with the infantile eczema can be obviously improved after 3 days, the eczema can be effectively eliminated after 5 days, and the infantile eczema is not easy to relapse after recovery.

2. Therapeutic effect on diaper dermatitis of infants

The follow-up clinical pediatrics is used for trial treatment of 22 patients with diaper dermatitis, the age is 2-16 months, the skin external ointment (dermatitis is preferred) prepared in example 1 is adopted, after defecation is ordered, warm water is used for washing the buttocks of the patients, a soft cleaning towel is used for absorbing water, then the external ointment is applied, parents are ordered to change diapers frequently, after the diapers are used for 2 days, congestion or red swelling of the skin of all the patients is relieved, and after the diapers are continuously used for 1 week, all the patients are cured. The effective rate of the external ointment for treating the diaper dermatitis of the infants is 100 percent, the external ointment has quick response, the hyperemia or the red and swollen symptoms of the skin of the infants can be obviously improved after 2 days, and the infants can be cured after 7 days of continuous use.

The above is only a preferred embodiment of the present invention, and it is not intended to limit the scope of the invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention shall be included in the scope of the present invention.

Claims

1. The external therapeutic agent for treating infantile eczema and/or diaper dermatitis is characterized by comprising an external composition, wherein the external composition consists of recombinant human metallothionein and resveratrol, the addition amount of the recombinant human metallothionein in the external therapeutic agent is 0.01-0.06 g/kg, and the addition amount of the resveratrol in the external therapeutic agent is 1-10 g/kg;

wherein the recombinant human metallothionein is any one of recombinant human metallothionein-1 and subtype and mutant thereof, and fusion protein of the recombinant human metallothionein-1; or the like, or, alternatively,

2. The external therapeutic agent according to claim 1, wherein the amount of the recombinant human metallothionein added to the external therapeutic agent is 0.02 to 0.05 g/kg;

3. The external therapeutic agent according to claim 2, wherein the recombinant human metallothionein is added in an amount of 0.025 to 0.04 g/kg;

4. The external therapeutic agent according to claim 1, which is a gel, an emulsion, a cream, a solution, a liniment, a lotion or an ointment.