KR20140111198A - Pharmaceutical composition for preventing or treating infectious diseases of skin comprising 5-Aminolevulinic acid - Google Patents

Abstract

The present invention relates to a pharmaceutical composition comprising 5-aminolevulinic acid or a derivative thereof as an active ingredient for preventing or treating skin injections, a cosmetic composition comprising 5-aminolevulinic acid or a derivative thereof as an active ingredient for alleviating skin infections, and a method for promoting expressions of IL-12 by using 5-aminolevulinic acid and functional cosmetics containing a cosmetic composition for alleviating skin infections. The composition comprising 5-aminolevulinic acid as an active ingredient of the present invention shows an effect of indirectly treating skin infections without side effects by promoting the secretion of IL-12 by acting on dendritic cells in the skin tissue, not on causative organism of skin injections, thus can be widely used for effectively preventing, treating, or alleviating skin infections.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a pharmaceutical composition for preventing or treating skin infections including 5-aminolevulinic acid,

The present invention relates to a pharmaceutical composition for preventing or treating a skin infectious disease comprising 5-aminolevulinic acid. More particularly, the present invention relates to a pharmaceutical composition for prevention or treatment of skin infections comprising 5-aminolevulinic acid or a derivative thereof as an active ingredient, A cosmetic composition for treating skin infections, comprising a pharmaceutical composition for therapeutic use, 5-aminolevulinic acid or a derivative thereof as an active ingredient, a functional cosmetic composition containing the cosmetic composition for improving skin infections, and 5-aminolevulinic acid, 12 < / RTI >

Human skin is the most important organ that has the biochemical and physical functions to protect the inside of the human body. The skin can be divided into three parts, epidermis, dermis, and hypodermis. It is a typical example of aging phenomena such as wrinkles, stains, and black spots due to ultraviolet rays, wounds, Symptoms appear.

In particular, various human skin diseases are known to be caused by microorganisms in particular hospitals. Depending on the type of infectious microorganisms, bacterial infections such as acne, acne, atopy, and fungal infections such as athlete's foot, ringworm, Respectively. These skin diseases are very diverse in the area of onset and onset, and in particular opportunistic and systemic infections, which may pose a life threat, include long-term use of anticancer drugs, use of immunosuppressive drugs due to organ or bone marrow transplantation, Which is caused by a decrease in the immune system in the body due to an increase in the number of the deficient patients and the aging of the population.

Especially known as a typical skin infectious disease, impetigo is a shallow purulent infection usually occurring in summer or in the skin of infants or infants. It is caused by blister impetigo and impetigo contagiosa It appears in the form of branches. Staphylococcus aureus is the main causative organism, and bacteria invade through areas of the skin. And skin irritation caused by skin infections are very common in pre-school children and are highly contagious and easily spread around. It often occurs everywhere in the arms, legs, face, and torso, and it is often scratched by itchy symptoms, which often spread to various parts of the body and cause new lesions to continue to develop. Usually, there are no systemic symptoms, but severe complications are rare, resulting in sepsis, pneumonia, and meningitis.

Conventionally, a chemical disinfectant such as Triclocarban or Triclosan has been used for sterilizing the causative bacteria of skin infections. However, it causes not only side effects such as induction of resistant bacteria and skin irritation, but also Gram negative bacteria and fungi It has a disadvantage in that the effect is relatively weak and its use is limited.

In order to overcome these drawbacks, it has been proposed to use polyenes such as amphotericin B, nystatin, natamycin, fluconazole, ketoconazole, itraconazole, cilofungin, chitin synthesis inhibitor (polyoxine, nikomycin) 5-fluorocytosine) has been developed, however, due to its wide use of antibiotics, frequent occurrence of resistant bacteria, irregular absorption, low pharmacokinetics and low specificity of chemical fungicide, the side effects are minimized and used for various hospital microbiological infections The development of a new therapeutic agent for skin infections is urgently required.

Under these circumstances, the inventors of the present invention have found that 5-Aminolevulinic acid (5-aminolevulinic acid), which is used in PDT (Photo Dynamic Therapy) therapy for treating acne, acid, ALA) or a derivative thereof stimulates dendritic cells present in skin cells to promote secretion of interleukin 12, thereby treating skin infections with secreted IL-12. Thus, the present invention has been completed.

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating skin infections comprising 5-aminolevulinic acid or a derivative thereof as an active ingredient.

Another object of the present invention is to provide a cosmetic composition for improving skin infections, which comprises 5-aminolevulinic acid or a derivative thereof as an active ingredient.

It is still another object of the present invention to provide a functional cosmetic composition containing the cosmetic composition for improving skin infections.

Another object of the present invention is to provide a method for promoting the expression of interleukin 12 comprising the step of treating 5-aminolevulinic acid to dendritic cells.

In one embodiment, the present invention provides a pharmaceutical composition for preventing or treating skin infections comprising 5-aminolevulinic acid or a derivative thereof as an active ingredient.

The term "5-aminolevulinic acid (ALA)" of the present invention means a biosynthesis of 5-aminolevulinic acid (ALA) by succinyl- CoA and glycin with 2-amino- Means a compound represented by the chemical formula of C ₅ H ₉ NO ₃ and having a molecular weight of 131.13, and is represented by the following formula.

The two molecules of 5-aminolevulinic acid are dehydrated and condensed by the phosphobilogenogenesis enzyme to form the phospholipids, which are phospholipids such as heme, picobiline, bile pigment, cyanocobalamine and the like. And it is known that 5-aminolevulinic acid undergoes an oxidative deamination reaction to give 2-oxoglutaric acid semialdehyde, which is then returned to succinic acid. Typically, the ALA is used in PDT (Photo Dynamic Therapy) therapy, which is known as acne therapy. In PDT therapy, active oxygen is produced when light is applied to porphyrin produced by acne bacterium (Coproporphyrin III) It is a method to treat acne by destroying acne bacterium by oxygen. At this time, the ALA is used as a porphyrin precursor to show an effect of doubling the efficiency of the PDT therapy. However, in order to perform the PDT therapy, light should be irradiated to the acne area. If the irradiation amount of light is excessive, active oxygen is excessively generated, which may damage not only acne bacteria but also skin cells. The inventors of the present invention conducted various studies to develop a solution to this problem and found that the ALA stimulates the dendritic cells distributed in the skin tissue and promotes the secretion of interleukin 12 from the dendritic cells. Respectively. The secreted interleukin 12 is a substance produced in dendritic cells contained in the skin tissue of a patient, minimizing side effects and not damaging skin cells. The inventors of the present invention firstly confirmed the effect of ALA.

The term " derivative of 5-aminolevulinic acid (ALA) "of the present invention means a compound having a structure in which the carboxyl group or amino group of ALA is substituted with another substituent and exhibits the same or similar activity as ALA. The derivative of ALA may be a compound including a structure in which the hydrogen atom of the carboxyl group of ALA or the chloride group of the amino group is substituted with another substituent, and more preferably, the hydrogen atom of the carboxyl group of ALA is a methyl group (-CH ₃ ), the b-ALA in which the hydrogen of the carboxy group of ALA is substituted with the butyl group (-C ₄ H ₉ ), the hydrogen of the carboxyl group of ALA is replaced by the hexyl group (-C ₆ H ₁₃ ) Substituted h-ALA, N-formyl-ALA in which the chlorinated group of the amino group of ALA is substituted with formyl group (-CHO), and so on, so long as the ALA derivative exhibits the same or similar activity as the ALA, But are not limited to compounds.

The term "dendritic cell" of the present invention refers to a cell in the form of a twig in the interstitial tissue spaces of various tissues including lymphoid tissues. It is derived from hematopoietic stem cells of bone marrow, Of the antigen-presenting cells. The dendritic cells express Class I and Class II histocompatibility antigens on the surface and are known to play an important role in T cell activation in the immune response. For the purpose of the present invention, the dendritic cell is used as a cell that overexpresses and secretes IL-12 by stimulation of ALA, but the use of the dendritic cell is not particularly limited thereto and may be used as a mediator of various immunotherapy have.

The term " interleukin 12 (IL-12) "of the present invention refers to a kind of interleukin that is a cytokine protein having biological activity that recognizes and produces antigens such as lymphocytes and macrophages. Means a glycoprotein having a molecular weight of about 70,000 produced. The IL-12 is in the form of a heterozygote (p70) in which two subunits of 40 kDa (p40) and 35 kDa (p35), which are not homologous to each other, are bound to one disulfide bond, Inducing activity of NK cells, stimulating cell proliferation by lectin stimulation of stably T cells, promoting differentiation into naive TH1 cells, promoting proliferation of hematopoietic stem cells in the presence of interleukin 3, And in vivo antitumor effects on transplant tumors. In the present invention, IL-12 is expressed in dendritic cells stimulated by ALA, and can induce the death of causative bacteria of skin infections caused by natural killer cells and the like.

The term "infectious diseases of skin" of the present invention means various diseases which are symptomatic due to infection of an exogenous bacterium such as a bacterium, fungus, rickettsia and spiroheta. The skin infectious disease is not particularly limited, but it may be infectious emetic which causes Staphylococcus aureus, skin mucositis causing fungi belonging to the fungus muchomo, and skin actinomycosis caused by anaerobic actinomycetes.

According to one embodiment of the present invention, when 5-aminolevulinic acid is treated to induce the activity of dendritic cells, the interleukin 12 protein is increased to have an anti-inflammatory effect in the dermal layer of the skin, thereby enhancing skin immunity , And that it can promote the differentiation of dendritic cells or T cells by dendritic cells.

Meanwhile, the pharmaceutical composition for preventing or treating skin infectious disease comprising ALA of the present invention as an active ingredient may further comprise an appropriate carrier, excipient or diluent commonly used in the production of a pharmaceutical composition. Examples of carriers, excipients and diluents that can be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. It is preferable to formulate a preparation for parenteral administration rather than an oral administration preparation because it directly acts on the skin. The parenteral administration preparation may contain a sterilized aqueous solution, a non-aqueous solution, a suspension, an emulsion, a lyophilized preparation, May be included. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

Particularly, it is preferable that the composition is formulated in the form of an external preparation for skin. The external preparation for skin is not particularly limited, but is preferably an ointment, a lotion, a spray, a patch, a cream, a powder, Can be manufactured and used in the form of a gel.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the drug form, the administration route and the period, but can be appropriately selected by those skilled in the art. However, for the desired effect, the composition is preferably administered at a daily dose of 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg. The administration may be carried out once a day or divided into several times.

The composition can be administered to mammals, including humans, in a variety of routes. All modes of administration may be expected, for example, by application to the affected area of the skin infections, or by muscle or subcutaneous injection.

As another embodiment for achieving the above object, the present invention provides a cosmetic composition for improving skin infectious disease comprising 5-aminolevulinic acid or a derivative thereof as an active ingredient.

Since the above-mentioned 5-aminolevulinic acid or its derivative acts directly on dendritic cells existing in the skin tissue to indirectly prevent or treat skin infectious diseases, the 5-aminolevulinic acid is useful as a cosmetic for improving skin infections May be used as an active ingredient of the composition.

The cosmetic composition for improving skin infections comprising 5-aminolevulinic acid or a derivative thereof of the present invention as an active ingredient may further comprise an additive. The kind of the additive is not particularly limited, but may be at least one of, for example, oil, water, a surfactant, a moisturizer, a lower alcohol, a bactericide, an antioxidant, a thickener, a chelating agent, a pigment, a preservative and a perfume. These additives may be used in an appropriate amount as required.

As another embodiment for accomplishing the above object, the present invention provides a functional cosmetic comprising the cosmetic composition for skin infectious disease improvement.

The functional cosmetic composition of the present invention exhibits an effect of improving skin infections and the formulation thereof is not particularly limited. For example, the functional cosmetics may be formulated into solutions, emulsions, suspensions, pastes, creams, lotions, gels, powders, The composition may be formulated into various forms such as an oil, a soap, a liquid cleansing agent, a bathing agent, a foundation, a makeup base, an essence, a lotion, a foam, a pack, a flexible water, a sunscreen cream, , Nutritional lotion, nutritional cream, massage cream, essence, pack, emulsion or oil gel.

For example, the ointment for external use may be prepared to contain 50 to 97% by weight of petrolatum and 0.1 to 5% by weight of polyoxyethylene oleyl-ether phosphate in addition to the above composition. For example, the softening water may be prepared to contain from 1 to 10% by weight of polyhydric alcohols such as propylene glycol or glycerin and from 0.05 to 2% by weight of a surfactant such as polyethylene oleyl ether or polyoxyethylene hydrogenated castor oil, The lotion and nutritional cream may be prepared to contain 5 to 20% by weight of an oil such as squalane, vaseline or octyldodecanol and 3 to 15% by weight of a wax component such as cetanol, stearyl alcohol or beeswax in addition to the above composition, The essence may be prepared to contain 5 to 30% by weight of polyhydric alcohols such as glycerin or propylene glycol.

Massage creams may be prepared in addition to the above composition to contain 30 to 70% by weight of oils such as liquid paraffin, petrolatum or isononylisononanoate, and the packs may contain peel (s) containing 5 to 20% by weight of polyvinyl alcohol off pack, and may be prepared as a wash off pack containing 5 to 30% by weight of a pigment such as kaolin, talc, zinc oxide or titanium dioxide in a general emulsified cosmetic.

In another embodiment for achieving the above object, the present invention provides a method for promoting the expression of interleukin 12 (IL-12) comprising the step of treating 5-aminolevulinic acid (5-ALA) .

The 5-ALA of the present invention not only activates dendritic cells but also increases the expression level of IL-12 in dendritic cells, and IL-12, which has an increased expression amount, exhibits normal activity, Treatment with cells can promote IL-12 expression.

The composition comprising 5-aminolevulinic acid as an active ingredient of the present invention acts on dendritic cells in skin tissue, which is not causative of skin infections, and promotes the secretion of IL-12, thereby indirectly treating skin infections , It can be widely used for effective prevention, treatment or improvement of skin infectious diseases since it does not show side effects.

1 is a graph showing the result of evaluation of toxicity of 5-aminolevulinic acid to leukocytes.
Fig. 2 shows the effect of dendritic cell activation of 5-aminolevulinic acid.
3 is a graph showing the promoting effect of cytokine synthesis of 5-aminolevulinic acid.
Fig. 4 is a photomicrograph showing the effect of promoting differentiation of 5-aminolevulinic acid into natural killer cells.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  1: 5- Of aminolevulinic acid (5-ALA)  Toxicity assessment

Mononuclear cells were extracted from the peripheral and spleen of BALB / c mice, and the extracted mononuclear cells were mixed with Histopak Ficoll and centrifuged at 800 rpm to obtain white blood cells. After 24 hrs of treatment with the various concentrations of 5-ALA, the resultant white blood cells were evaluated for MTT assay (Fig. 1). 1 is a graph showing the result of evaluation of toxicity of 5-aminolevulinic acid to leukocytes. As shown in FIG. 1, it was found that treatment with 5-ALA up to 10 μM did not show toxicity to leukocytes.

Example  2: 5- ALA Of dendritic cell activation

The white blood cells obtained by the method of Example 1 were further subjected to high-speed centrifugation to obtain dendritic cells contained in the white blood cells. The dendritic cells obtained were treated with various concentrations of 5-ALA and cooled in ice water for 15 minutes. The cooled cells were washed twice with 1500 rpm, fixed with 2% formalin, fixed dendritic cells were stained with complement CD30, and fluorescent dyed dendritic cells were analyzed using a flow cytometer (Fig. 2). Fig. 2 shows the effect of dendritic cell activation of 5-aminolevulinic acid. As shown in FIG. 2, when 10 μM of 5-ALA was treated, dendritic cells were activated.

Example  3: 5- ALA Promoting the synthesis of cytokines

10 [mu] M of 5-ALA or 50 [mu] g / ml of IL-12 were each treated with dendritic cells obtained by the method of Example 2 and centrifuged at 2,000 rpm after 24 hours to obtain supernatant . The obtained supernatant was diluted 3-fold and applied to an ELISA kit for quantification of interferon gamma or an ELISA kit for quantification of IL-12 to compare changes in expression levels of interferon gamma or IL-12 (FIG. 3). 3 is a graph showing the promoting effect of cytokine synthesis of 5-aminolevulinic acid. As shown in FIG. 3, the expression level of interferon gamma and IL-12, which are cytokines, was remarkably increased when 5-ALA was treated. Especially, interferon gamma increased the expression level of 5-ALA compared with IL- And the amount of water was relatively increased.

Example  4: 5- ALA Of natural killer cells ( NK cell ) Differentiation promoting effect

From the results of Example 3, it was confirmed that when dendritic cells were treated with 5-ALA, the expression level of IL-12 was increased. Therefore, it was examined whether or not IL-12 having the increased expression level showed normal activity. For this purpose, the following experiments were carried out using the properties of promoting the maturation of natural killer cells (NK cells) among the various activities of IL-12 in white blood cells.

Specifically, the leukocytes obtained by the method of Example 1 were treated with 10 μM of 5-ALA, and after 24 hours, Giemsa staining, which is used for dyeing blood cells, was performed. Gamma staining stains the white nucleus nucleus in purple or purple. Natural killer cells show normal monocyte morphology when they are not mature, but after mature by treatment with IL-12 and the like, they are atypical and polynuclear (Fig. 4). As a result, it was confirmed whether or not the maturation of natural kill cells was promoted through the staining of Kimja (Fig. 4). Fig. 4 is a photomicrograph showing the effect of promoting differentiation of 5-aminolevulinic acid into natural killer cells. As shown in FIG. 4, it was confirmed that the 5-ALA treatment promoted the maturation of natural killer cells. Thus, it was found that IL-12 having increased expression level by 5-ALA treatment exhibited normal activity.

The results of Examples 1 to 4 show that 5-ALA does not show toxicity to dendritic cells but activates dendritic cells as well as increases the expression level of IL-12 in dendritic cells and increases IL- 12 showed normal activity. Thus, 5-ALA acts on dendritic cells to promote the expression and secretion of IL-12 and to kill the causative bacteria of skin infections by secreted IL-12 I could.

Claims (12)

A pharmaceutical composition for preventing or treating skin infections comprising 5-aminolevulinic acid (ALA) or a derivative thereof as an active ingredient.
The method according to claim 1,
The derivatives of 5-aminolevulinic acid (ALA) include m-ALA in which the hydrogen of the carboxy group of ALA is substituted with a methyl group (-CH ₃ ), b of the carboxy group of ALA substituted with a butyl group (-C ₄ H ₉ ) -ALA, with the hydrogen of the carboxyl group of ALA hexyl group (-C ₆ h ₁₃₎ substituted h-ALA, the N-formyl-ALA and combinations thereof optionally substituted with a formyl group (-CHO) chloride of the amino group of ALA by Lt; / RTI > is a compound selected from the group consisting of.
The method according to claim 1,
Wherein the skin infectious disease is a disease selected from the group consisting of infectious emetic, skin mucositis, cutaneous actinomycosis, and combinations thereof.
The method according to claim 1,
Wherein the composition further comprises a pharmaceutically acceptable diluent, excipient or carrier.
The method according to claim 1,
And stimulating the dendritic cells in the skin tissue to promote the secretion of interleukin 12 from the dendritic cells.
5-aminolevulinic acid or a derivative thereof as an active ingredient.
The method according to claim 6,
Wherein the composition further comprises at least one member selected from the group consisting of water, a surfactant, a moisturizer, a lower alcohol, a chelating agent, a bactericide, an antioxidant, an antiseptic, a pigment and a perfume.
The method according to claim 6,
Wherein the skin infectious disease is a disease selected from the group consisting of infectious emetic, skin mucositis, cutaneous actinomycosis, and combinations thereof.
The method according to claim 6,
The derivatives of 5-aminolevulinic acid (ALA) include m-ALA in which the hydrogen of the carboxy group of ALA is substituted with a methyl group (-CH ₃ ), the hydrogen of the carboxy group of ALA is substituted with a butyl group (-C ₄ H ₉ ) b-ALA, the hydrogen of the carboxyl group of ALA hexyl group (-C ₆ h ₁₃₎ as a substituted h-ALA, the N-formyl-ALA and combinations thereof optionally substituted with a formyl group (-CHO) chloride of the amino group of ALA ≪ / RTI > is a compound selected from the group consisting of:
9. A functional cosmetic for improving skin infections comprising the composition of any one of claims 6 to 9.
9. The method of claim 8,
A lotion, a gel, a powder, a spray, a surfactant-containing cleansing oil, a soap, a liquid cleansing agent, a bath agent, a foundation, a makeup base, an essence, a lotion, a foam, Sunscreen cream or sun oil. ≪ RTI ID = 0.0 > 18. < / RTI >
A method of promoting the expression of interleukin 12 (IL-12) in dendritic cells, comprising the step of treating dendritic cells with 5-aminolevulinic acid (5-ALA).

Images