KR20180059340A - Composition for skin-whitening comprising non thermal plasma treated solution - Google Patents

Abstract

The present invention relates to a composition for skin-whitening comprising liquid plasma. More specifically, the present invention relates to a cosmetic composition for skin-whitening comprising liquid plasma as an active ingredient, and a pharmaceutical composition for treating or preventing skin pigmentation diseases.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for skin whitening comprising a liquid-phase plasma,

The present invention relates to a skin whitening composition comprising a liquid-phase plasma. More specifically, the present invention relates to a skin whitening cosmetic composition containing a liquid plasma as an active ingredient, and a pharmaceutical composition for treating or preventing skin pigmentation diseases.

Skin melanogenesis is a defense mechanism against stimuli such as ultraviolet light in melanocyte-producing cells (melanocyte), and a large amount of melanin produced by the melanin is transferred to keratinocyte and accumulated on the skin surface layer Results. Although melanin protects the skin, hyperpigmentation of the skin causes skin irritation, freckles, darkening of the skin after inflammation of the skin and aging pigmentation spots. This causes the discomfort of the person not only the cosmetic discomfort but also the psychological negative affect on the social activities. . Melanin is produced by tyrosine tyrosine, an enzyme called tyrosinase, converted into DOPA or dopaquinone, and then enzymatically oxidized to form an abnormal deposit in the skin And it is known that it produces spots and black spots. Materials for inhibiting melanin production such as hydroquinone, arbutin, vitamin C and derivatives thereof have been developed for the relief, prevention and treatment of such pigment deposits, stains and spots, and whitening cosmetics containing them have been disclosed And Korean Patent Publication No. 2005-0509848 discloses a whitening cosmetic composition containing a keractone derivative isolated from Korean Patent Publication No. 2005-0509848. Korean Patent Publication No. 2005-0479741 discloses a whitening cosmetic containing a glucose acylated derivative. Of these, hydroquinone is generally recognized for its effectiveness, but is generally limited in its use because of its sensitivity. Ascorbic acid is easily oxidized, the cosmetic compounded with it causes discoloration and irregularities, and the substances derived from plant extracts are difficult to maintain the homogeneity of the product due to the great difference in the effect according to the plant area of the plant, and glucose acylated derivatives The synthesis efficiency is very low.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide a skin whitening composition containing a liquid plasma.

Accordingly, the present inventors have made intensive efforts to develop a method for the fundamental treatment of skin pigmentation, and as a result, it has been confirmed that the liquid plasma can be used for skin whitening because it inhibits tyrosinase activity and inhibits melanin biosynthesis And completed the present invention.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, for purposes of complete understanding of the present invention, various specific details are set forth, such as specific forms, compositions, and processes, and the like. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well-known processes and techniques of manufacture are not described in any detail, in order not to unnecessarily obscure the present invention. Reference throughout this specification to "one embodiment" or "embodiment" means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in at least one embodiment of the invention. Accordingly, the appearances of the phrase " in one embodiment "or" an embodiment "in various places throughout this specification are not necessarily indicative of the same embodiment of the present invention. In addition, a particular feature, form, composition, or characteristic may be combined in any suitable manner in one or more embodiments.

It is an object of the present invention to provide a skin whitening composition comprising a liquid plasma which exhibits excellent whitening effect through inhibition of tyrosinase activity and inhibition of melanin biosynthesis, and which is also excellent in stability.

Another object of the present invention is to provide a skin whitening cosmetic composition containing the skin whitening composition containing the liquid plasma.

It is still another object of the present invention to provide a pharmaceutical composition for treating or preventing a pigmentation disease containing the plasma-containing skin whitening composition.

As used herein, the term "whitening" refers to the ability of the skin to be continuously exposed to ultraviolet light to increase melanocytes and thereby prevent excessive deposition of melanin pigment in the skin, or to thin existing melanin pigment . As a result, it is possible to inhibit spots and freckles caused by excessive deposition of melanin pigment.

As used herein, the term " non-thermal plasma treated solution " (NTS) refers to the generation of a high-density, high energy plasma in a liquid and may be produced by exposure to atmospheric pressure nonthermal plasma . The term " liquid-phase plasma " can be used interchangeably with the term " plasma-conditioned liquid material " It is water, saline, buffer, or medium, and most preferably a medium.

As used herein, the term "culture media" refers to a medium that enables cell growth and survival in vitro to be supported, and includes conventional media used in the art suitable for culturing cells All included. Depending on the type of cells, medium and culture conditions can be selected. The basic medium used for culturing the cells is preferably a cell culture minimum medium (CCMM), and generally includes a carbon source, a nitrogen source and a trace element component. These cell culture basal media include, for example, DMEM (Dulbecco's Modified Eagle's Medium), MEM (Minimal Essential Medium), BME (Basal Medium Eagle), RPMI1640, F-10, F- Glasgow's Minimal Essential Medium), Iscove's Modified Dulbecco's Medium, and the like.

In one embodiment of the present invention, "treatment " means any action that improves or alleviates skin pigmentation and other conditions associated therewith using liquid-phase plasmas according to the present invention. If you are a knowledgeable person, you can refer to the data provided by the Korean Medical Association, etc. to determine the exact criteria for skin pigmentation, and to judge the degree of improvement, improvement and treatment.

In one embodiment of the present invention, " prevention " means any action that inhibits or delays skin pigmentation and other conditions associated therewith using a liquid plasma according to the present invention. It will be apparent to those skilled in the art that compositions of the present invention that are therapeutically effective in skin pigmentation disorders can prevent such diseases using liquid-phase plasmas according to the present invention before the initial symptoms or symptoms of skin pigmentation appear.

As used herein, the term " pharmaceutical composition " means a composition to be administered for a specific purpose. For the purpose of the present invention, the pharmaceutical composition of the present invention comprises a liquid plasma prepared by irradiating a plasma with a liquid substance, and may include a protein involved therein and a pharmaceutically acceptable carrier, excipient or diluent have. The above "pharmaceutically acceptable" carrier or excipient as referred to above means approved by the regulatory agency of the government or listed in government or other generally approved pharmacopoeias for use in vertebrates, and more particularly in humans do.

For parenteral administration, the pharmaceutical compositions of this invention may be in the form of a suspension, solution or emulsion in an oily or aqueous carrier, and may be prepared in the form of a solid or semi-solid. In addition, the pharmaceutical compositions of the present invention may include a formulating agent such as a suspending, stabilizing, solubilizing and / or dispersing agent and may be sterilized. The pharmaceutical composition may be stable under the conditions of manufacture and storage and may be preserved against the contaminating action of microorganisms such as bacteria or fungi. Alternatively, the pharmaceutical compositions of the invention may be in the form of a sterile powder for reconstitution with suitable carriers before use. The pharmaceutical compositions may be presented in unit-dose form, in micro needle patches, in ampoules, or in other unit-dose containers, or in multi-dose containers. Alternatively, the pharmaceutical composition may be stored in a freeze-dried (lyophilized) condition requiring only the addition of a sterile liquid carrier, e. G., A vehicle immediately prior to use. Immediately, the injection solutions and suspensions may be prepared from sterile powders, granules or tablets.

In some non-limiting embodiments, the pharmaceutical compositions of the present invention may be formulated, or may be included in the form of microspheres in a liquid. In certain non-limiting embodiments, the pharmaceutical compositions of the present invention may include pharmaceutically acceptable compounds and / or mixtures thereof at a concentration of between 0.001 and 100,000 U / kg. Also, in certain non-limiting embodiments, the pharmaceutical compositions of the present invention may contain suitable excipients such as preserving agents, suspending agents, additional stabilizing agents, dyes, buffers, antimicrobial agents, antifungal agents, and isotonic agents such as, for example, . As used herein, the term "stabilizer" refers to a compound that is optionally used in the pharmaceutical compositions of the present invention to increase shelf life. In a non-limiting embodiment, the stabilizing agent may be a sugar, an amino acid, or a polymer. The pharmaceutical composition of the present invention may also include one or more pharmaceutically acceptable carriers, which may be solvents or dispersion media. Non-limiting examples of pharmaceutically acceptable carriers include water, saline, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycols), oils, and suitable mixtures thereof. Non-limiting examples of sterilization techniques applied to the pharmaceutical compositions of the present invention include filtration through a bacteria-inhibiting filter, sterilization of the terminals, incorporation of a sterile preparation, irradiation, sterilization gas irradiation, heating, vacuum drying and freeze drying do.

As used herein, " administration " means introducing the composition of the present invention to a patient by any appropriate method, and the administration route of the composition of the present invention may be administered through any conventional route so long as it can reach the target tissue have. Intravenous, intraperitoneal, intraperitoneal, intradermal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intraperitoneal, intradermal, intradermal, intraocular, The composition is most preferably, but not exclusively, provided in the form of being applied on the skin or injected subcutaneously or intradermally.

The method of treatment of the present invention may comprise administering the pharmaceutical composition in a pharmaceutically effective amount. In the present invention, the effective amount may be determined depending on the kind of the disease, the severity of the disease, the kind and amount of the active ingredient and other ingredients contained in the composition, the type of the formulation and the age, body weight, general health condition, sex and diet, And the fraction of the composition, the duration of the treatment, the drug being co-administered, and the like.

As used herein, the term " cosmetic composition " is a composition for improving or improving skin pigmentation and other symptoms thereof. The cosmetic composition comprising the composition of the present invention as an active ingredient can be used as a skin lotion, a nutritional lotion, a nutrition essence, a massage cream, a cosmetic bath additive, a body lotion, a body milk, a bath oil, a baby oil, Skin lotion, skin cream, sunscreen cosmetics, cleansing milk, depilatory (makeup), face and body lotion, face and body cream, skin whitening cream, hand lotion, hair lotion, Soap, shampoo, hand cleanser, medicinal soap (non-medical use), cream soap, facial wash, hair rinse, makeup soap, tooth whitening gel, toothpaste, etc. . ≪ / RTI > To this end, the compositions of the present invention may further comprise suitable carriers, excipients or diluents conventionally used in the preparation of cosmetic compositions. The carrier, excipient or diluent which may further be added to the cosmetic composition of the present invention include purified water, oil, wax, fatty acid, fatty alcohol, fatty acid ester, surfactant, humectant, thickener, antioxidant, Buffers, lower alcohols, and the like, but are not limited thereto. Further, if necessary, it may contain a whitening agent, a moisturizing agent, a vitamin, an ultraviolet screening agent, a perfume, a dye, an antibiotic, an antibacterial agent, and an antifungal agent. As the oil, hydrogenated vegetable oil, castor oil, cottonseed oil, olive oil, palm oil, jojoba oil and avocado oil may be used. Examples of the wax include wax, wax, carnauba, candelilla, montan, ceresin, liquid paraffin, Can be used. Examples of the fatty acid include stearic acid, linoleic acid, linolenic acid and oleic acid. The fatty acid alcohols include cetyl alcohol, octyldodecanol, oleyl alcohol, panthenol, lanolin alcohol, stearyl alcohol and hexadecanol As the fatty acid ester, isopropyl myristate, isopropyl palmitate, and butyl stearate may be used. As the surfactant, a cationic surfactant, an anionic surfactant and a nonionic surfactant known in the art can be used, and a surfactant derived from a natural material is preferably used. In addition, it may contain a hygroscopic agent, a thickening agent, an antioxidant and the like widely known in the field of cosmetics, and the kind and amount thereof are well known in the art.

In one embodiment of the present invention, there is provided a method of manufacturing a plasma processing apparatus, comprising the steps of: (a) filling a plasma generator with a carrier gas; (b) supplying a voltage of 0.5 kV to 20 kV and a frequency of 10 to 30 kHz to the plasma generator to generate a plasma; And (c) irradiating the generated plasma with a liquid substance, wherein the carrier gas in the step (a) is at least one selected from the group consisting of nitrogen, helium, argon, and oxygen Wherein the carrier gas is helium and oxygen, wherein the carrier gas is helium and oxygen, wherein the carrier gas is helium and oxygen, wherein the carrier gas is helium and oxygen, Is performed for 1 minute per 1 ml at a distance of 0.1 cm to 15 cm from the surface of the liquid material, characterized in that the liquid substance in step (c) There is provided a method for producing a liquid-phase plasma for skin whitening which is water, saline solution, buffer solution, or culture medium.

In another embodiment of the present invention, there is provided a skin whitening cosmetic composition comprising a liquid plasma prepared by any one of the above methods, wherein the cosmetic composition is at least one of skin lotion, skin softener, skin toner, astringent, lotion, Which is selected from the group consisting of moisturizing lotion, nutritional lotion, massage cream, nutritional cream, moisture cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion or body cleanser The present invention provides a cosmetic composition for skin whitening which is a single formulation.

According to another embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating skin pigmentation diseases, which comprises a liquid plasma prepared by any one of the above methods as an active ingredient, wherein the skin pigmentation disease is a skin pigment, Wherein the pigment is at least one selected from the group consisting of black spots, nevi, pigmentation by drugs, pigmentation after inflammation, hyperpigmentation occurring in dermatitis, and a pharmaceutical composition for preventing or treating skin pigmentation diseases Characterized in that the pharmaceutical composition is a cream, a gel, a patch, a spray, an ointment, an alarm, a lotion, an essence, a liniment, a pasta or a percutaneous absorbent preparation. A pharmaceutical composition is provided.

In another embodiment of the present invention, there is provided a method of preventing or treating a skin pigmentation disease comprising administering a pharmaceutical composition prepared by any one of the above methods to a subject other than a human, Is at least one selected from the group consisting of spots, freckles, black spots, nevi, pigmentation by drugs, pigmentation after inflammation, and hyperpigmentation caused by dermatitis. Provide a treatment method.

Hereinafter, the present invention will be described in detail.

The cosmetic composition for skin whitening comprising a liquid plasma according to the present invention and the composition for the treatment or prevention of pigmentation diseases exhibit an effect of inhibiting excellent tyrosinase activity and inhibiting melanin biosynthesis, so that the whitening cosmetic composition, skin pigmentation It is anticipated that it can be usefully used as a pharmaceutical composition for the treatment or prevention of diseases.

1 is a view showing a method of manufacturing a liquid-phase plasma (NTS) according to the present invention.
2 shows the effect of inhibiting pigment formation in a zebrafish of a liquid-phase plasma according to the present invention (Experimental Example 1).
FIG. 3 shows the effect of inhibiting pigment formation in a human tissue skin of a liquid-phase plasma according to the present invention (Experimental Example 2).
FIG. 4 shows the effect of inhibiting tyrosinase activity in Human Tissue skin of a liquid-phase plasma according to the present invention (Experimental Example 3).
FIG. 5 shows the results of the toxic effect test in the melanocytes of the liquid-phase plasma according to the present invention (Experimental Example 4).
FIG. 6 shows the effect of reducing the melanogenesis marker in human melanocytes of the liquid-phase plasma according to the present invention (Experimental Example 5).

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

<Examples>

Example 1. Preparation of liquid phase plasma (NTS)

10 ml of each of F12 (10% FBS 24 ug / ml 3-isobutyl-1-methylxanthine, 80 nM 12-O-tetradecanoyl Phorbor 13- acetate (TPA), 1.2 g / ml bFGF and 0.1 ug / ml cholera toxin) , And a liquid plasma ("plasma-treated medium") was completed using helium and oxygen as a carrier gas at a distance of 1 cm from the medium at a strength of 4 KV for 1 minute per ml. A method of producing the liquid-phase plasma is shown in Fig.

<Experimental Example>

Experimental Example  One. In Zebrafish  Pigment inhibition effect

The embryos collected from the zebrafish female and male embryos were placed in a zebrafish embryo culture medium (Egg water, 0.2 g sea slat / L) for 24 hours, and then the liquid plasma prepared in Example 1 was treated. In order to increase the degree of penetration of the chorion sample during the sample processing, corona of the embryo was pierced with tweezers. After 24 hours from the sample treatment, the embryos showing the toxicity of the sample were removed, After the complete removal, samples of the same concentration were treated again. After 48 hours of sample treatment, the extent of pigmentation in the sample-treated group as compared to the control group was observed by a stereo microscope, and the results are shown in FIG.

Referring to FIG. 2, it was confirmed that zebrafish treated with liquid plasma suppressed the formation of pigment compared to the control group.

Experimental Example 2. Inhibitory Effect of Pigmentation on Human Tissue Skin

Human foreskin tissue was cultured in Transwell (DMEM high glucose + 4% FBS), and the liquid plasma prepared in Example 1 was treated with 1 min / 1 ml. After 3 days of incubation, fixation was performed and the degree of pigment formation was observed through Fontana-masson stain. The results are shown in FIG.

Referring to FIG. 3, it was confirmed that the epidermal staining of the human Tissue skin treated with the liquid plasma was slightly lower than that of the control group.

Experimental Example 3 Inhibitory Effect of Tyrosinase Activity on Human Tissue Skin

The human foreskin sample was seeded in a 60 mm dish at 2 × 10 ⁶ , and then treated with 1 ml / 1 ml of the liquid plasma prepared in Example 1. After 24 hours of incubation, the cells were fixed and Western blot, PCR, tyrosinase activities and melanin content were measured. The results are shown in FIG.

As can be seen from FIG. 4, tyrosinase, MITF, and melanin content were significantly decreased in Human Tissue skin treated with liquid plasma.

Experimental Example 4. Toxic Effect Test in Melanocyte

The cytotoxicity of human melanocytes was confirmed by time after treatment with the liquid plasma prepared in Example 1, and the results are shown in FIG.

As can be seen from FIG. 5, it was confirmed that the liquid plasma according to the present invention does not show toxicity in human melanocytes.

Experimental Example 5. Melanogenesis marker reduction effect in human melanocyte

After the liquid plasma prepared in Example 1 was treated with human melanocytes, the reduction effect of the melanogenesis markers was confirmed over time, and the results are shown in FIG.

As can be seen from FIG. 6, the liquid plasma according to the present invention was confirmed to reduce melanogenesis markers in human melanocytes, and it was confirmed that not only the protein level but also the RNA level of tyrosinase was reduced.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (8)

(a) filling the plasma generator with a carrier gas;
(b) supplying a voltage of 0.5 kV to 20 kV and a frequency of 10 to 30 kHz to the plasma generator to generate a plasma; And
(c) irradiating the generated plasma with a liquid substance.
The method according to claim 1,
Wherein the carrier gas in the step (a) is at least one selected from the group consisting of nitrogen, helium, argon, and oxygen.
The method according to claim 1,
Wherein the liquid substance in step (c) is water, saline, a buffer, or a medium.
A cosmetic composition for skin whitening comprising a liquid plasma produced by the method according to any one of claims 1 to 3.
A pharmaceutical composition for preventing or treating skin pigmentation diseases, comprising a liquid plasma prepared by any one of claims 1 to 3 as an active ingredient.
6. The method of claim 5,
Wherein the skin pigmentation disorder is any one or more selected from the group consisting of spots, freckles, black spots, nevi, pigmentation by drugs, pigmentation after inflammation, and hyperpigmentation caused by dermatitis. A pharmaceutical composition for preventing or treating a pigmentation disorder.
A method for preventing or treating skin pigmentation diseases, comprising administering a pharmaceutical composition prepared by any one of claims 5 to 6 to a subject other than a human.
8. The method of claim 7,
Wherein the skin pigmentation disorder is any one or more selected from the group consisting of spots, freckles, black spots, nevi, pigmentation by drugs, pigmentation after inflammation, and hyperpigmentation caused by dermatitis. A method for preventing or treating a pigmentation disorder.

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