KR20140035565A - Chlorin e6 for the treatment, prevention or improvement of acne - Google Patents

Abstract

The present invention relates to a pharmaceutical composition or a cosmetic composition comprising a chlorin e6 as an active ingredient which has an outstanding antimicrobial activity for acne germs. Since the chlorin e6 has antimicrobial, anti-inflammatory, and antioxidant activities, the chlorin e6 can be used as an active ingredient for treating, preventing, and alleviating various skin diseases and, in particular, has an excellent effect on acne. [Reference numerals] (AA) Growth rate of acne germs (%); (BB) Fluorescent light irradiation; (CC) Darkroom; (DD) Control group; (EE) Test group (50 mM)

Description

Chlorin e6 for the treatment, prevention or improvement of acne}

The present invention relates to a pharmaceutical composition or cosmetic composition comprising chlorin e6, which is excellent in antibacterial activity against acne bacteria, as an active ingredient.

Chlorine e6 is well known as a photosensitizer used in photodynamic therapy (PDT) of malignancies. Photosensitizers require a characteristic that selectively accumulates only in tumor tissues, that is, selectivity in tumor tissues and normal tissues. Selective photosensitizers can increase the efficacy of PDT, shorten the treatment time, and reduce the side effects of drugs injected into the body. When irradiated with light of a specific wavelength to a photo-sensitizer, it generates single oxygen and radical species, which is a kind of active oxygen, which directly kills tumor cells and induces immunoinflammatory reactions. Or damage the tumor microvascular system. Conventional photosensitizers have been known to accumulate selectively to some extent in tumors, but some also accumulate in normal tissues including skin. However, chlorine e6 is known to have high selectivity for malignant cells and no toxicity to normal cells compared to other photosensitive agents used in tumor therapy.

As described above, chlorine e6 has already been reported in several documents that it is effective as an anticancer agent. [Korean Patent Registration No.808,630,841,959]

However, to date, no literature has reported that chlorine e6 has excellent antibacterial activity against acne.

In the present invention, to provide a pharmaceutical composition or cosmetic composition for the treatment, prevention or improvement of acne.

In order to solve the above problems, the present invention provides a pharmaceutical composition for treating and preventing acne wherein chlorin e6 is contained as an active ingredient.

The present invention also provides cosmetic compositions for preventing and improving acne, wherein chlorin e6 is contained as an active ingredient.

Most of the acne treatments on the market are synthesized antibiotics such as benzoyl peroxide, tetracycline, erythromycin and clindamycin, which have side effects and safety problems, but chlorine e6 is a natural product and has excellent human compatibility.

In addition, chlorin e6 has antibacterial, anti-inflammatory and antioxidative activities and thus is useful as an active ingredient for the treatment, prevention and improvement of various skin diseases, and particularly has excellent efficacy against acne.

In addition, since chlorine e6 is a photosensitizer activated by sunlight and incandescent lamps, chlorine e6 is applied to the skin and irradiated with natural light to destroy sebaceous glands and acne causing bacteria, thereby treating, preventing and improving acne.

1 is a graph confirming the efficacy of chlorine e6 inhibit the growth of acne bacteria when irradiated with an incandescent light source.

The present invention relates to a pharmaceutical composition or cosmetic composition for the purpose of treating, preventing or ameliorating acne in which chlorin e6 is contained as an active ingredient.

Chlorine e6 comprising the composition of the present invention as an active ingredient is a natural product separated and purified from chlorophyll extract, Korean Patent No. 1,180,695 discloses a method for separating and purifying high purity chlorine e6 from chlorophyll extract.

1. Preparation of pharmaceutical composition

The pharmaceutical composition of the present invention may be used as an active ingredient in chlorine e6 by further comprising a suitable carrier, excipient and diluent commonly used in the pharmaceutical manufacturing field as external preparations such as liquids, suspensions, ointments, patches and the like. Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of pharmaceutical preparations, diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, and surfactants are usually used.

The pharmaceutical composition of the present invention can be transdermally administered, and the preferable dosage of the pharmaceutical composition 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 of the present invention is preferably administered at 0.0001 to 1 g / kg per day, preferably 0.001 to 200 mg / kg per day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

2. Preparation of cosmetic composition

The cosmetic composition of the present invention may be in the form of an oil dispersion in an aqueous phase using an aqueous, aqueous-alcoholic or oily solution, oil or water or multi-emulsion, aqueous or oily gel, liquid, pasty or solid anhydrous products, And more preferably in the form of a lipid vesicle in ionic and / or non-ionic form.

The cosmetic composition of the present invention may be somewhat fluid and may have the appearance of a white or colored cream, ointment, milk lotion, serum, essence, paste or mousse. It may optionally be applied to the skin in aerosol form, or it may be in solid form, e.g., in the form of a stick. It may also be used as a skin care and / or makeup product.

In a known manner, the cosmetic composition according to the present invention may also contain conventional adjuvants such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, perfumes, fillers, And dyes. The amount of these various adjuvants is an amount ordinarily used in the art and ranges, for example, from 0.0001 to 10% by weight based on the total weight of the cosmetic composition. Depending on their nature, such adjuvants can be introduced into the lipid, aqueous phase, lipid vesicles and / or nanoparticles. In any case, the adjuvants and their proportions will be chosen so as not to adversely affect the desired properties of the cosmetic composition according to the invention.

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto.

[Example] Efficacy of chlorin e6

Example 1 Antimicrobial Activity of Chlorin e6

In order to measure the antimicrobial activity of chlorine e6, it proceeded according to the liquid culture inhibition method. Escherichia coli is cultured in LB liquid medium, Pseudomonas aeruginosa , Bacillus cereus , Staphylococcus aureus.subsp.aureus Staphylococcus epidermidis was cultured in nutrient liquid media. After culturing until the absorbance was 1 at 620 nm, cells were collected and used for the experiment.

In addition, Propionibacterium acnes , acne strain, was cultured in an anaerobic liquid medium for 4 days and diluted to 1000: 1 to measure the initial bacterial count. In order to measure the antimicrobial activity of each bacteria, 300 μl of each bacteria were collected after incubation, and the absorbance was measured at 620 nm using an ELISA reader. The growth rate of each microorganism was measured. After culturing each bacteria in the sample untreated medium, the absorbance value at this time was used as a control. The results are shown in FIG. 1.

Example 2. In vivo antibacterial test of chlorine e6

The experimental animals received male C57BL / 6 mice at 7 weeks of age of about 20 g body weight and were acclimated for 7 days. Experimental mice were divided into three groups and separated into groups to freely feed and water. Group A was injected with propionibacterium acnes (1 × 10 ⁷ CFU / 20 μL in PBS) in the left ear of the mouse and 20 μL of PBS in the right ear intradermally. The treatment group injected intradermal propionibacterium acnes (1 × 10 ⁷ CFU / 20 μL in PBS) into both ears of the mouse, and then applied various concentrations of samples to the right ear. Mice in each group were sacrificed with cervical vertebrae 24 hours after the start of the experiment, and ear tissues were extracted.

The extracted ear tissue was excised and fixed in 10% formalin, and subjected to a series of tissue treatments, and then embedded in a paraffin embedding machine (Leica EG116) to prepare 4 μm thick sections using a microtome (Leica 820), followed by H & E (Hematoxylin). & Eosin) staining was observed by light microscopy.

Example 3. Measurement of changes in fat accumulation in sebaceous gland cells by chlorine e6

SZ95 cells, which are human immortalized sebocytes, were cultured in a medium supplemented with 10% fetal bovine serum (FBS, Gibco RBL Co.) in a CO ₂ incubator (37 ° C, 5% CO ₂ ) Co. Germany).

SZ95 cells were cultured on a chamber slide, stimulated with testosterone, and treated with chlorin e6 in a concentration-dependent manner. After the culture was completed, the cells were stained with oil red, stained with Harris hematoxylin, and observed for Lipid droplet in SZ95 cells by optical microscope.

SZ95 cells were dispensed into 1 × 10 ⁵ cells in a 100Φ dish and incubated for 48 hours, followed by incubation for 5 days in a sample in sebocyte serum-free medium for 5 days. The cultured cells were separated and collected, and then the cells were lysed by solubilization with a solution buffer (lysis buffer; EDTA 10 μL, 100 mM SPB solution, PMSF 1 μL, Triton X-100 10 μL) and the protein was quantified. A chloroform: methanol (2: 1) solution was added to the cell lysate, and centrifuged at 2000 rpm for 15 minutes to collect the lower layer solution. Secondarily, a chloroform: methanol (2: 1) solution was added to the remaining solution by centrifugation, and the lower layer liquid was separated in the same manner as above to concentrate with nitrogen gas to extract lipids.

To the extracted lipid was added 300 μL of ethanol and dissolved. Then, 50 μL of cell lipid was added to H ₂ SO ₄ , boiled for 10 minutes, and then cooled at 4 ° C. to form a ketone body. 100 μL of the cooled sample was added to 6 mL of phospho-vanillin reagent, treated at 37 ° C for 15 min, and absorbance was measured at a wavelength of 540 nm. The standard value was 1 to 50 mg / mL.

Example 4 Determination of Cholesterol Content Changes in Sebaceous Gland Cells by Chlorin e6

Cholesterol in the sebaceous gland cells was measured using a total cholesterol kit after dissolving 300 μL of ethanol into the extracted lipid. 100 μL of the sample was added to 1 mL of the enzyme solution, incubated at 37 ° C. for 5 minutes, and the absorbance at 500 nm was measured. Cholesterol in the standard solution was adjusted to 3 mg / mL by adding 20 μL of the standard solution to 3 mL of the enzyme solution.

Example 5 Measurement of Triglyceride Contents in Sebaceous Gland Cells by Chlorin e6

300 μL of ethanol was added to the extracted lipid, and the triglyceride in sebaceous gland cells was measured using a triglyceride kit. 100 μL of the sample was added to 1 mL of the enzyme solution, and the mixture was incubated at 37 ° C. for 5 minutes. The absorbance was measured at a wavelength of 546 nm. Triglyceride of the standard solution was added to 3 mL enzyme solution 20 μL of the standard solution was set to 3 mg / mL value.

[Production of medicines]

Pharmaceuticals 1. Preparation of emulsion

0.5 g of chlorin e6, 5 g of stearyl alcohol, 0.8 g of polysorbate-60, 4 g of cyclomethicone, 2 g of jojoboyl, 0.2 g of methyl paraben, 8 g of 1,3-butylene glycol, 0.05 g of xanthan gum, 1.2 g of polyacrylamide / C13-14 isoparaffin / laureth-7, 0.2 g of imidazolidinyl urea, 0.15 g of triethanolamine, 0.1 g of perfume, and the amount of water were weighed. The oil-based raw materials among the above-mentioned composition components were dissolved by heating in an oil-phase auxiliary tank at 75 占 폚. The water-based raw materials among the above-mentioned composition ingredients were dissolved by heating in an emulsification tank at 75 占 폚. The emulsion tank was charged with the emulsified raw material under vacuum and emulsified for 5 minutes at a temperature of 75 ° C at 2500 rpm and a pedal mixer at 3500 rpm using a homogenizer. After degassing under vacuum, the emulsion was cooled to obtain an emulsion preparation.

2. Preparation of patches

15 g of glycerin, 2 g of polyarylic acid, 2 g of acrylate copolymer, 0.4 g of ammonium hydroxide, 0.05 g of disodium-EDTA, and 0.2 g of tartaric acid were added to a dissolution tank and homogeneously stirred at room temperature to prepare a viscous liquid. 0.2 g of methylparaben, 1 g of ethanol, 0.1 g of dipotassium glycyrrhizinate, and 0.05 g of perfume were dissolved at room temperature, injected into the dissolution tank, and then homogeneously stirred. 0.5 g of chlorin e6, 5 g of propylene glycol, and purified water were dissolved at 60 DEG C and then injected into the dissolution tank in small amounts to prepare an attachment gel.

The attached gel prepared above was coated on the siliconized polyester film using a suitable coating machine with an applicator gap, and then the polyethylene film was laminated. Then, it was put in a pouch laminated film composed of paper, low density polyethylene and aluminum to prepare an attachment.

Pharmaceuticals 3. Manufacture of liquids

100 mg of chlorin e6, 10 g of isomerized sugar and 5 g of mannitol were dissolved in purified water in accordance with the usual preparation method of the liquid preparation, and the components were added and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, Was added with purified water to adjust the total volume to 100 ml, and sterilized to prepare a liquid preparation.

[Production of cosmetic preparation]

Cosmetics 1. Production of lotion

30 mg of chlorin e6, 5 mg of polyoxyethylene hardened pharmacopoeial, 30 mg of glycine, 1 mg of dipotassium glycyrrhizate, 30 mg of 1,3-butylene glycol, 1 mg of sodium hyaruronate, 50 mg of ethanol, 1 mg of triethanolamine, 1 mg of triethanolamine, 1 mg of EDTA, and purified water were mixed to prepare a lotion.

Cosmetics 2. Manufacture of Lotion

Chlorine e6 15 mg, L-ascorbic acid-2-magnesium phosphate salt 100 mg, water-soluble collagen (1% aqueous solution) 100 mg, sodium citrate 10 mg, citric acid 5 mg, 1,3-butylene glycol 300 mg, purified water The lotion was prepared by mixing.

Cosmetics 3. Manufacture of cream

1 mg of glycine, 6 mg of chlorin e6, 200 mg of polyethylene glycol monostearate, 500 mg of self-emulsifying monostearate glycerin, 400 mg of cetyl alcohol, 600 mg of squalane, 600 mg of tri-2-ethylhexane glyceryl, 700 mg of 3-butylene glycol, and purified water were mixed to prepare a cream.

Claims (8)

A pharmaceutical composition for the treatment and prevention of acne wherein chlorin e6 is contained as an active ingredient.
The method of claim 1,
Wherein the pharmaceutical composition is formulated with an external preparation for skin selected from the group consisting of a liquid preparation, an emulsion, a suspension, an ointment, and an adjuvant.
The method of claim 1,
Wherein the chlorin e6 is a photosensitizer which is activated by sunlight or a fluorescent lamp to generate active oxygen species.
The method of claim 1,
A pharmaceutical composition characterized by treating and preventing acne by PhotoDynamic Therapy.
A cosmetic composition for preventing and improving acne wherein chlorin e6 is contained as an active ingredient.
The method of claim 5, wherein
A lotion, a cream, a lotion, and a cream.
The method of claim 5, wherein
Wherein the chlorin e6 is a photosensitizer which is activated by sunlight or a fluorescent lamp to generate active oxygen species.
The method of claim 7, wherein
A cosmetic composition characterized by preventing and improving acne by photodynamic therapy.

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