KR101716850B1 - Composition for whitening containing proteins derived from mesenchymal stromal cells - Google Patents

Abstract

The present invention relates to a whitening cosmetic composition and a whitening food composition containing, as an active ingredient, a protein having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells. The present invention also relates to a pharmaceutical composition for preventing or treating diseases caused by hyperpigmentation, which comprises a protein having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells as an active ingredient. The present invention also provides a method for producing a mesenchymal stem cell having a whitening activity, comprising the steps of extracting a protein from mesenchymal stromal cells and isolating the extracted protein by size to obtain a protein having a size of 5 kDa to 1,000 kDa And a method for producing the resulting protein. According to the present invention, proteins having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells can inhibit melanin to a high level in melanin-producing cells and thus can be used for various whitening applications.

Description

TECHNICAL FIELD [0001] The present invention relates to a whitening composition containing a mesenchymal stem cell-derived protein,

The present invention relates to a whitening cosmetic composition and a whitening food composition containing, as an active ingredient, a protein having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells. The present invention also relates to a pharmaceutical composition for preventing or treating diseases caused by hyperpigmentation, which comprises a protein having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells as an active ingredient. The present invention also provides a method for producing a mesenchymal stem cell having a whitening activity, comprising the steps of extracting a protein from mesenchymal stromal cells and isolating the extracted protein by size to obtain a protein having a size of 5 kDa to 1,000 kDa And a method for producing the resulting protein.

Melanin is produced against the damage of the skin caused by ultraviolet rays and protects the skin. The process of biosynthesis of melanin begins with the oxidation of tyrosine by tyrosinase. After the production of DOPA (3,4-dihydroxyphenylalanine) and dopachrome, DHICA (5,6 -Dihydroxyindole-2-carboxylic acid) and DHI (5,6-dihydroxyindole), and it is known that DHI-eumelanin, DHICA-eumelanin, and peomelanin are produced.

Specifically, melanin synthesis is mediated by the action of tyrosinase and tyrosinase related proteins present in the melanosome. Tyrosine is oxidized by tyrosinase to form DOPA, and then DOPA is oxidized again by the action of dopaoxidase to generate dopaquinone. Then, tyrosinase-related protein TRP-1 (5, 6-dihydroxyindole-2-carboxylic acid oxidase) and TRP-2 (dopachrome tautomerase). The expression of tyrosinase, TRP-1 and TRP-2 is promoted by a transcription factor MITF (microphthalmia-associated transcription factor). MITF is inhibited when ERK (extracellular signal regulated kinase) is activated. Therefore, a whitening effect can be expected when the expression of tyrosinase and / or MITF is inhibited or the expression of ERK is promoted.

Blackening of the skin occurs when melanin produced in melanocytes in skin cells is transferred to keratinocytes and accumulates in skin surface layer. Melanin pigmentation and deposition of the skin cause spots, freckles, and black spots. Kojic acid, arbutin, vitamin C derivative and the like have been proposed as a whitening agent in order to prevent and improve the blackening of such skin.

However, in the case of kojic acid, it has been reported that the inhibitory effect on melanogenesis in cell and in vivo is less than that of in vitro inhibition of tyrosinase, and that liver cancer can be induced during long-term use. In addition, vitamin C derivatives have a problem of chemical stability, which makes it difficult to store them for a long period of time. As the side effects of whitening active materials are known, studies on functional whitening materials as alternative raw materials have been actively conducted.

On the other hand, stem cells are undifferentiated cells that can divide for a long time through self-renewal and can differentiate into various kinds of cells under specific circumstances. Stem cells are classified into embryonic stem cells and adult stem cells depending on the origin of the stem cells. In terms of research using the stem cells, the potential of adult stem cells is limited to embryonic stem cells However, since there are few ethical problems, many studies have been conducted on adult stem cells.

Mesenchymal stromal cells (MSCs) are pluripotent undifferentiated cells, also called mesenchymal stem cells. Mesenchymal stromal cells can be differentiated into various types of cells including osteoblasts, chondroblasts, adipocytes, and the like.

Korean Patent Registration No. 10-0848056 discloses a method for inhibiting melanin synthesis using an adipose-derived stem cell culture medium, which is an mesenchymal stem cell, and Korean Patent Laid-Open Publication No. 10-2009-0116659 discloses a method for inhibiting melanin synthesis in a cord blood derived mesenchymal stem cell culture ≪ RTI ID = 0.0 > cosmetic < / RTI >

It is an object of the present invention to provide a new whitening composition exhibiting optimal whitening activity by using proteins extracted from stem cells, in particular, mesenchymal stromal cells.

Accordingly, the present invention has been accomplished in order to achieve the above object, and as a result, it has been found that a protein having a specific size range extracted from mesenchymal stromal cells has excellent whitening activity, and thus the present invention has been completed.

According to the present invention, proteins having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells can inhibit melanin expression in melanin-producing cells to a high level and thus can be used for various whitening applications.

FIG. 1 shows a sample for inhibition of melanin formation according to an embodiment of the present invention.
FIG. 2 is a graph showing the inhibition rate of melanin formation according to an embodiment of the present invention. FIG.
3 is a graph showing the melanin formation inhibition rate of the embodiment of the present invention and the melanin formation inhibition rate of mesenchymal stromal cells (hereinafter referred to as SRP2 protein) exceeding 1,000 kDa according to the concentration.
FIG. 4 is a graph showing the inhibition rate of melanin production according to an embodiment of the present invention derived from various cells.

Hereinafter, the present invention will be described in detail. However, it should be understood that the present invention may be embodied in many other specific forms without departing from the spirit or essential characteristics thereof.

In one aspect, the invention relates to a whitening composition using stem cells, particularly mesenchymal stromal cells.

The present invention relates to a whitening cosmetic composition containing, as an active ingredient, a protein having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells.

The present invention also relates to a whitening food composition comprising as an active ingredient a protein having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells.

The present invention is also applicable to pharmaceutical compositions for the treatment or prevention of diseases caused by hyperpigmentation of the skin. Hyperpigmentation refers to the symptoms of darkening or darkening of the skin or nail compared to other areas due to excessive increase of melanin in certain areas of the nail. Diseases caused by hyperpigmentation include, but are not limited to, spots, freckles, aging pigmentation, sunburn, and the like.

In another aspect, the present invention provides a method for producing a protein, comprising: a) extracting a protein from mesenchymal stromal cells, and b) isolating the extracted protein by size to obtain a protein having a size of 5 kDa to 1,000 kDa. Lt; RTI ID = 0.0 > a < / RTI > mesenchymal cell-derived protein.

The mesenchymal stromal cells of the present invention are preferably human mesenchymal stromal cells extracted from human fat, placenta, umbilical cord blood or bone marrow. In addition, human embryonic stem cells can be used as mesenchymal stem cells after differentiation into mesenchymal cells. As an example, the mesenchymal stromal cells of the present invention use adult stem cells isolated from adult adipocytes, which can be obtained by simple purification of cells present in adipose tissue.

Proteins having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells in the composition of the present invention may be contained in an amount of 0.001% to 20% based on the total weight of the composition, It can be adjusted appropriately according to the content. However, if the content of the protein having a size of 5 kDa to 1,000 kDa extracted from the mesenchymal stromal cells in the composition is less than 0.001%, it may be difficult to expect a substantial whitening effect. If the content is more than 20% Lt; / RTI > Therefore, it is preferable that the content is in the range of 0.001% to 20% based on the total weight of the composition.

In one embodiment of the present invention, the composition of the present invention has a skin whitening effect so as to enhance or reinforce the skin whitening effect, in addition to the protein having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells as an active ingredient Or other known compounds or natural extracts that are known or will be known in the future.

In one embodiment of the present invention, the cosmetic composition of the present invention may further contain at least one cosmetically acceptable additive. The at least one additive that is acceptable for cosmetics is not particularly limited and may be selected from conventional ingredients incorporated in common cosmetics such as oil, water, a surfactant, a moisturizer, an alcohol, a thickener, a colorant, an antiseptic, a flavor, a chelating agent, , Sunscreen agents, fatty acids, and the like.

The cosmetic composition of the present invention may be any one selected from the group consisting of a skin, a lotion, a cream, a gel, an essence, a foundation, a lotion, a patch, a pack, a soap, a cleanser and a detergent without particular limitation.

In one embodiment of the invention, the food composition of the present invention may additionally contain one or more additives which are acceptable for food use. The at least one additive that is acceptable for foodstuffs is not particularly limited and may contain conventional ingredients that can be incorporated into foods, such as water, flavor, flavor, colorant, stabilizer, preservative, alcohol, Vitamins, plant extracts, organic acids, minerals, nutrients, and the like.

The form of the food composition of the present invention is not particularly limited, but may be any form selected from the group consisting of beverage, gum, tea, confectionery, vitamin complex and health supplement.

In one embodiment of the invention, the pharmaceutical composition of the present invention may further contain one or more pharmaceutically acceptable carriers. The pharmaceutically acceptable carriers are not particularly limited and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, Polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., in addition to the above carrier.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, the mode of administration, the condition of the patient, and the severity of symptoms, but it is preferable to administer 0.01 mg / kg to 2,000 mg / kg per day. The pharmaceutical composition of the present invention can be administered orally or parenterally, and when administered parenterally, it can be administered by application, intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, or the like. For example, it may be desirable to administer the pharmaceutical composition of the present invention in a topical application to the skin, since it is used in the treatment of skin disorders due to hyperpigmentation.

In the present invention, skin whitening-related activities of proteins having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells were measured. As a result, it was confirmed that melanin - producing cells exhibit higher melanin production inhibitory activity than arbutin, a whitening functional substance.

The present invention will be described in more detail with reference to the following examples. However, the following examples are for the purpose of illustrating the present invention and are not intended to limit the scope of the present invention.

[ Example ]

Intermediate lobe From stromal cells  Protein extraction

Derived from human adipose tissue, extracted from human bone marrow, extracted from human umbilical cord, and human embryonic stem cells differentiated and extracted into mesenchymal cells. 1 × 10 ⁶ mesenchymal stromal cells were collected and pulsed off for 15 seconds using ultrasonic wave crusher (VCX500; manufactured by SONICS & MATERIALS INC., USA) at 4 ° C and then pulsed off for 5 seconds. And the amplitude power was set to 20%, and the ultrasonic wave was repeated three times within one minute. The supernatant was centrifuged at 15,000 g for 5 minutes to remove the supernatant and the pellet was removed.

Separation of extracted proteins by size

Vivaspin filtration tubes (manufactured by Vivascience, UK) and Amicon filtration tubes (manufactured by Millipore, USA) equipped with polyethersulfone membranes containing pore sizes of 1,000 kDa, 500 kDa, 100 kDa and 5 kDa cut- ) Were used to separate the extracted proteins by size.

Measurement of inhibitory activity of melanin formation

The inhibitory rate of melanin formation in melanin-producing cells was measured in a protein having a size of 5 kDa to 1,000 kDa (hereinafter referred to as 'SRP1 protein').

Melanogenesis cells (manufactured by CEFO, Korea) extracted from human skin tissue were used. The melanin-producing cells were cultured using CEFOgro MELplus medium (manufactured by CEFO, Korea). In order to examine the ability of the SRP1 protein to inhibit melanogenesis, Comparative Example 1 (negative control) was cultured with CEFOgro MELplus and Comparative Example 2 (positive control) was cultured with CEBSgro MELplus culture supplemented with 2 mM arbutin. The gelatin-concentrated SRP1 protein was mixed and coated on a 12-well cell culture dish (manufactured by Corning Incorporated, USA), and the melanocytes were seeded and cultured with CEFOgro MELplus. All the examples and comparative examples were cultured for 72 hours, and the absorbance at 405 nm was measured after dissolving the cells with 1 N NaOH.

As shown in Fig. 1, the examples showed melanin inhibitory activity to the extent that they could be confirmed even by the naked eye. The results of the absorbance measurement are shown in FIG. 2, and it was confirmed that when the control group of Comparative Example 1 was set to 100%, the inhibition of melanin formation was remarkably higher than that of Comparative Example 2, which was a positive control group treated with arbutin.

In addition, as shown in FIG. 3, the SRP1 protein inhibited melanin production in a concentration-dependent manner. On the contrary, the SRP2 protein, which is over 1,000 kDa, showed almost no melanin production inhibitory activity.

FIG. 4 also shows that mesenchymal stromal cells (ADMSC) extracted from human adipose tissue, mesenchymal stromal cells (BMMSC) extracted from human bone marrow, mesenchymal stromal cells (UCMSC) extracted from human umbilical cord, It is shown that SRP1 protein derived from mesenchymal stromal cells (ESMSC) derived after differentiation of cells into mesenchymal cells shows melanin formation inhibitory activity.

Claims (8)

A cosmetic composition for skin whitening comprising, as an active ingredient, a protein having a size of 5 kDa to 1,000 kDa extracted from mesenchymal stromal cells disrupted by ultrasonication. The method according to claim 1,
Wherein the mesenchymal stromal cells are human mesenchymal stromal cells extracted from human fat, placenta, umbilical cord blood or bone marrow. delete delete delete delete A method for producing a mesenchymal stem cell-derived protein having a whitening activity comprising the steps of:
a) extracting proteins from mesenchymal stromal cells by ultrasonication; And
b) separating the extracted proteins by size to obtain a protein having a size of 5 kDa to 1,000 kDa. 8. The method of claim 7,
Wherein the mesenchymal stromal cells are human mesenchymal stromal cells extracted from human fats, placenta, umbilical cord blood or bone marrow.

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