KR102216302B1 - A composition for activating a skin cell and uses thereof - Google Patents

Abstract

The present specification relates to a composition for activating skin stem cells, and the composition may include at least one component of ferrous sulfate and lipoic acid.
The composition provided herein may exhibit an activating effect of skin cells, and the skin cells may be skin stem cells.
Stem cell proliferation and/or division occurs actively by the skin stem cell activation, and ultimately, aged skin can be regenerated.
In addition, skin whitening effects may also appear by the composition.

Description

A composition for activating a skin cell and uses thereof

The content disclosed by the present specification relates to a composition for activating skin cells, and more specifically, to a composition for activating skin stem cells, including ferrous sulfate and lipoic acid.

In recent years, interest in stem cell cosmetics is increasing, and general stem cell cosmetics refer to cosmetics containing as a component a stem cell culture solution containing components secreted during stem cell culture.

Cosmetics including such a stem cell culture medium as well as an ingredient capable of increasing the activity of the stem cells themselves are being developed, and in this case, there is an advantage of being able to manufacture cosmetics through a low-cost raw material and a simple process.

An example to be achieved by the content disclosed by the present application is to provide a composition for activating skin cells.

Another example to be achieved by the content disclosed by the present application is to provide a cosmetic composition for activating skin cells.

In order to achieve the above-described problem, according to an aspect of the technology disclosed by the present application, ferrous sulfate; And lipoic acid (Lipoic acid) or a salt thereof; a composition for activating skin cells is provided.

The ferrous sulfate may be ferrous sulfate (iron (II) sulfate; FeSO ₄ ), for example, tetrahydrate (FeSO ₄ ·4H ₂ O), pentahydrate (FeSO ₄ ·5H ₂ O ), 6 hydrated salts (FeSO ₄ ·6H ₂ O) and 7 hydrated salts (FeSO ₄ ·7H ₂ O).

In addition, the lipoic acid may be alpha-lipoic acid or beta-lipoic acid, and the alpha-lipoic acid is (R)-(+)-alphalipoic acid. ((R)-(+)-α-lipoic acid), (S)-(-)-alpha-lipoic acid ((S)-(-)-α-lipoic acid), or a mixture thereof.

The skin may be derma or epiderma, and cells activated by the composition may be fully differentiated cells, progenitor cells, or stem cells. .

In addition, in the composition for activating skin cells, in addition to ferrous sulfate and lipoic acid or salts thereof, which act as main components, amino acids, sugars, vitamins, inorganic salts, lipids, cytokines, growth factors ( Growth factor), an extracellular matrix protein, a medium, an antioxidant, a melanin synthesis inhibitory substance, a toxic substance acting on cells including a melanin pigment, and a keratin exfoliation substance.

As described above, the composition for activating skin cells may function i) a cell proliferation effect and/or inhibit expression of a cell senescence regulator. The aging modulator is a CDK inhibitor (Cyclic-dependent Kinase (CDK) inhibitor), and specifically, it is characterized in that at least one of P21, P27, P57, P16, P15, P18, P19, P53 and p-P53.

According to another aspect of the technology disclosed by the present application, ferrous sulfate; And Lipoic acid or a salt thereof; There is provided a cosmetic composition for skin cell activation comprising one or more of.

The cosmetic composition is Glycine, Arginine, L-Glutamine, L-Glutamic Acid, asparagine (L-Asparagine-H2O), and alanine (L-Alanine). ), Aspartic acid, L-Cysteine hydrochloride-H2O, Histidine, L-Isoleucine, L-Leucine, L-Lysine ), methionine (L-Methionine), phenylalanine (L-Phenylalanine), serine (L-Serine), threonine (L-Threonine), tryptophan (L-Tryptophan), proline (L-Proline), valine (L-Valine) , D-Calcium pantothenate, Folic Acid, Niacinamide, Pyridoxine hydrochloride, Riboflavin, Thiamine hydrochloride, Cyanocobalamine (Vitamin B12), Ascorbic Acid, Biotin, Inositol, Calcium Chloride (CaCl2), Magnesium Sulfate (MgSO4)), Potassium Chloride (KCl)), Sodium Bicarbonate (NaHCO3), Sodium Chloride (NaCl)), Sodium Phosphate monobasic (NaH2PO4-H2O)), Glucose (D-Glucose (Dextrose)), Sodium It may further include one or more of pyruvate (Sodium Pyruvate), adenosine (Adenosine), and guanosine (Guanosine).

The above cosmetics are softening lotion, nutritional lotion, lotion, skin, body lotion, nutrition cream, massage cream, moisture cream, hand cream, eye cream, essence, cleansing foam, cleansing cream, cleansing water, pack, gel, patch, lipstick, Makeup base, powder, foundation, body cleanser, toothpaste, mouthwash, shampoo, conditioner, hair tonic, gel, mousse, hair essence such as hair essence, it may be a formulation selected from the group consisting of hair dye and hair dye.

The cosmetic composition may exhibit i) anti-aging effect, ii) regeneration effect, iii) wrinkle improvement effect, iv) whitening effect and/or v) skin elasticity increase effect.

Cell activation effects may be exhibited by the composition disclosed by the present specification, and ultimately skin condition improvement effects may also appear.

The cell activation effect may be a cell proliferation, differentiation, or cell senescence inhibitory effect. In addition, as a skin condition improvement effect, wrinkle improvement effect, anti-aging effect, skin elasticity effect, and whitening effect may appear.

1 is a human fibroblast (Human Primary) by Mix II and Mix III in which ferrous sulfate and alpha-lipoic acid are included in a modified medium and other components are included in a modified medium according to an embodiment. Fibroblast) proliferative effect.
Figure 2 (a) shows the degree of proliferation of human primary fibroblasts by a composition containing alpha-lipoic acid (α-Lipoic acid).
Figure 2 (b) shows the degree of proliferation of human primary fibroblasts by a composition containing ferrous sulfate.
3 shows the proliferation effect of human primary fibroblasts by compositions containing ferrous sulfate and α-Lipoic acid (Experimental Examples (a) to Experimental Examples (i)).
FIG. 4 shows the results of confirming the dermal stem cell markers (CD29(+), CD34(-)) of Holoclone and the proliferation effect of Holoclone.
5 shows the results of confirming the dermal stem cell markers (CD44(+), CD90(+), CD105(+)) of a holoclone.
6 shows the degree of proliferation of holoclone by a composition containing ferrous sulfate.
7 shows the degree of proliferation of holoclone by a composition containing α-Lipoic acid.
8 shows the effect of proliferation of holoclones by compositions containing ferrous sulfate and α-Lipoic acid (Experimental Examples (1) to (9)).
9 is a graph comparing the proliferation effect of human primary fibroblasts and holoclones according to Experimental Example (8).
10 shows the degree of proliferation of bone marrow-derived mesenchymal stem cells according to Experimental Examples (1) to (9).
11 shows the degree of proliferation of fat-derived mesenchymal stem cells according to Experimental Examples (1) to (9).
12 shows the wound healing effect of the compositions Mix I, Mix II and Mix III.
13 shows the effect of inhibiting the expression of aging markers (p-P53, P21) according to Experimental Example (8).
14 shows the degree of proliferation of human primary fibroblasts by a composition containing L-Aspartic acid, L-Alanine, or Proline in DMEM basic medium.
15 is a human primary fibroblast proliferation by a composition containing pantothenic acid Ca-salt, Pyridoxine hydrochloride or L-Ascorbic acid in DMEM basic medium Indicates the degree.
16 is a human fibroblast (Human) by a composition containing L-asparagine monohydrate, L-glutamic acid, or D-biotin in DMEM basic medium Primary Fibroblast) indicates the degree of proliferation.
17 shows the degree of proliferation of human primary fibroblasts by a composition containing zinc sulfate heptahydrate or riboflavin in DMEM basic medium.

Definition of Terms

Definitions of representative terms used in the present specification are as follows.

The term "skin" is an organ composed of tissues that protect organs and muscles in the body from external stimuli, and can be damaged by various external factors, and the skin is composed of the epidermis, the dermis and the subcutaneous fat layer. The epidermis (Epidermis) ) Consists of the stratum corneum, granule layer, play layer, and base layer.

The epidermis includes Keratinocytes, Melanocytes, Langerhans cells, and Merkels cells, and Keratinocytes are the majority of epidermal cells (about 95%). Occupy.

The dermis is a layer of skin that exists between the epidermis and the subcutaneous fat layer, and includes extracellular matrix proteins such as collagen and elastin and fibroblasts.

The subcutaneous fat layer (Hypodermis) is the thickest layer of the skin and consists of adipose tissue and connective tissue. The subcutaneous fat layer contains fat cells.

The subcutaneous fat layer (Hypodermis) is the thickest layer of the skin and consists of adipose tissue and connective tissue. The subcutaneous fat layer contains fat cells.

The term "fibroblast" is a type of cell that synthesizes the extracellular matrix and collagen, and functions to secrete a precursor of the extracellular matrix to maintain the structural integrity of the connective tissue.

The term "Primary cell" refers to a cell cultured separately from living tissue.

The term "Human Primary Fibroblast" refers to fibroblasts isolated and cultured from human tissue.

The term "stem cell" refers to an undifferentiated cell that has the ability to self-reproduce through replication and differentiate into various cells. For example, there are embryonic stem cells or adult stem cells.

The term "adult stem cell" refers to an undifferentiated cell existing between differentiated cells of a specific tissue or organ, and may serve to replace and repair damaged tissue. Adult stem cells can only differentiate into individual target organs.

The term "skin stem cell" is a type of adult stem cell and is present in a very small amount in skin cells. For example, skin stem cells include epidermal stem cells, hair follicle stem cells, or dermal stem cells.

The term “Holoclone” is a stem cell that forms a colony and is well known to those skilled in the art. (Nature Medicine volume 12, pages 1397-1402 (2006))

In the present specification, the term "holoclone" and the term "stem cell" may be used interchangeably.

The term “skin cells” refers to cells derived from the skin. For example, the skin cells may include, but are not limited to, fibroblasts, skin stem cells, and progenitor cells.

The term "medium" is a medium containing nutrients for culturing cells, and the medium may be a basic medium or a modified medium. For example, the medium may be a solid medium or a liquid medium, but is not limited thereto.

The term "culture" refers to proliferation or maintenance of the cells, and the culture may include both adherent culture and suspension culture.

The term “proliferation” may mean that cells are divided and the number of cells increases, and the proliferation of the cells includes both proliferation into a single layer or multilayer.

The term "basic medium" is a commercially available medium for culturing cells, and may be a known medium commonly used for culturing cells.

For example, the basic medium may be a DMEM medium, MEM, BME, RPMI 1640, F-10, F-12, or α-MEM medium, but is not limited thereto.

The term "modified medium" means a medium in which some of the constituents of the basic medium are modified.

For example, the modified medium may be when some of the constituents of the basic medium have been removed. For another example, the modified medium may be a case in which components other than the constituents of the basic medium are added to the constituent components of the basic medium. For another example, the modified medium may be when some of the constituents of the basic medium are removed and other ingredients other than the constituents of the basic medium are added.

The term “about” refers to 30, 25, 20, 25, 10, 9, 8, 7, 6, 5, or 30, 25, 20, 25, 10, 9, 8, 7, 6, 5, or 2 for a reference amount, level, value, number, frequency, percentage, dimension, size, amount, weight or length. It means an amount, level, value, number, frequency, percentage, dimension, size, amount, weight or length varying by 4, 3, 2 or 1%.

In addition to these terms, other terms are defined elsewhere in the specification as necessary. Unless clearly defined otherwise herein, industry terms used herein will have industry-recognized meanings.

Hereinafter, the content disclosed by the present specification will be described in detail.

Main components of the composition for cell activation

One content disclosed by the present specification includes a composition for cell activation.

The composition may contain at least one of ferrous sulfate and lipoic acid as an essential component.

The ferrous sulfate is Green vitriol, Iron vitriol, Coppers, Melanterite, Szomolnokite, Iron sulfate, Iron sulphate, Ferrous sulfate. (Ferrosulfate), FeSO4, may be referred to as green salts (Green salts).

The ferrous sulfate is ferrous sulfate (iron (II) sulfate; FeSO ₄ ), ferrous sulfate ferric sulfate (iron (II) iron sulfate (III); FeSO ₄ Fe ₂ (SO ₄ ) ₃ ) Or ferric sulfate (iron (III) sulfate; Fe ₂ (SO ₄ ) ₃ ).

In one embodiment, in the case of ferrous sulfate, anhydrous salt (FeSO ₄ ), tetrahydrate (FeSO ₄ ·4H ₂ O), pentahydrate (FeSO ₄ ·5H ₂ O), hexahydrate (FeSO ₄ ·6H ₂ O) ) Or 7 hydrated salt (FeSO ₄ ·7H ₂ O).

In another embodiment, in the case of ferric sulfate, an anhydrous salt (Fe ₂ (SO ₄ ) ₃ ), a trihydrate (Fe ₂ (SO ₄ ) ₃ ·3H ₂ O), hexahydrate (Fe ₂ (SO ₄ ) ₃ 6H ₂ O), 7 hydrated (Fe ₂ (SO ₄ ) ₃ 7H ₂ O), 7.5 hydrated (Fe ₂ (SO ₄ ) ₃ 7.5H ₂ O), 9 hydrated (Fe ₂ (SO ₄ ) ₃ 9H ₂ O), 10 hydrated salt (Fe ₂ (SO ₄ ) ₃ ·10H ₂ O) or 12 hydrated salt (Fe ₂ (SO ₄ ) ₃ ·12H ₂ O).

As a representative example, ferrous sulfate may be 7 hydrated salt (FeSO ₄ ·7H ₂ O).

A person skilled in the art may optionally adjust the content of ferrous sulfate and lipoic acid to include in the composition, and in this specification, the content in the composition is expressed as a molar concentration (mol/L; M). I can.

A person skilled in the art may optionally adjust the content of ferrous sulfate and include it in the composition.

For example, ferrous sulfate may be included in an amount of about 0.5 μM to 1 mM in the composition.

In one embodiment, ferrous sulfate may be included in the range of about 0.5 μM to 100 μM.

For example, about 0.5 μM to 1 μM of ferrous sulfate may be included. For example, about 1 μM to 50 μM of ferrous sulfate may be included. For example, about 50 μM to 100 μM of ferrous sulfate may be included.

In another embodiment, ferrous sulfate may be included in about 100 μM to 1 mM.

For example, ferrous sulfate may be included in the range of about 100 μM to 500 μM. For example, ferrous sulfate may be included in about 500 μM to 1 mM.

In the present specification, ferrous sulfate, which may be included as a major component, is generally known as a material that supplies oxygen or supplements iron.

Lipoic acid is a fatty acid containing 8 carbons and 2 sulfur, a substance produced in a small amount in the human body, and the amount of production in the body decreases with aging.

The lipoic acid may be alpha-lipoic acid or beta-lipoic acid, but is not limited thereto.

The alpha-lipoic acid (α-Lipoic acid) is 5-(dithioren-3-yl)pentanoic acid (5-(dithiolan-3-yl)pentanoic acid), thioctic acid, DL-alphalipoic acid ( DL-α-Lipoic acid), 6,8-Thioctic acid, Lipothion, Liposan, Thioctsan, C ₈ H ₁₄ O ₂ S ₂ Or it may be referred to as Espa-lipon.

In one embodiment, alpha-lipoic acid (α-Lipoic acid) may be pure enantiomers or a mixture thereof. For example, (R)-(+)-alpha-lipoic acid ((R)-(+)-α-Lipoic acid), (S)-(-)-alpha-lipoic acid ((S)-(-)-α- Lipoic acid) or mixtures thereof.

In another embodiment, alpha-lipoic acid (α-Lipoic acid) may be a derivative or salt thereof.

The beta-lipoic acid (β-Lipoic acid) is 5-[(1R,3R)-1-ketodithiore-3-yl]valeric acid (5-[(1R,3R)-1-ketodithiolan-3-yl] valeric acid), 5-[(1R,3R)-1-oxo-1,2-dithioren-3-yl]pentanoic acid (5-[(1R,3R)-1-oxo-1,2-dithiolan -3-yl]pentanoic acid) or C ₈ H ₁₄ O ₃ S ₂ .

In one embodiment, beta-lipoic acid (β-Lipoic acid) may be pure enantiomers (pure enantiomers) or a mixture thereof. For example, beta-lipoic acid (β-Lipoic acid) is (R)-(+)-beta-lipoic acid ((R)-(+)-β-Lipoic acid), (S)-(-)-beta-lipoic acid (( S)-(-)-β-Lipoic acid) or mixtures thereof.

One of ordinary skill in the art may optionally adjust the concentration of the lipoic acid and include it in the composition.

For example, the lipoic acid may be included in an amount of about 0.5 μM to 1 mM in the composition.

In one embodiment, lipoic acid may be included in about 0.5 μM to 100 μM.

For example, lipoic acid may be included in about 0.5 μM to 50 μM. For example, lipoic acid may be included in about 50 μM to 100 μM. In a specific example, about 0.5 μM of lipoic acid may be included.

In another embodiment, lipoic acid may be included in the range of about 100 μM to 300 μM.

For example, lipoic acid may be included in the range of about 100 μM to 250 μM. In a specific example, about 200 μM of lipoic acid may be included. For example, lipoic acid may be included in the range of about 250 μM to 300 μM.

In another embodiment, lipoic acid may be included in about 300 μM to 500 μM.

For example, about 300 μM to 450 μM of lipoic acid may be included. For example, about 400 μM of lipoic acid may be included. For example, lipoic acid may be included in about 450 μM to 500 μM.

In another embodiment, lipoic acid may be included in the range of about 500 μM to 1 mM.

Lipoic acid, which may be included as a major component in the present specification, is generally known as a substance that functions as a coenzyme and antioxidant that regulates metabolism in the body.

For example, lipoic acid serves to break down fat, activate mitochondria, promote the synthesis of antioxidants in the body, or reactivate antioxidants in the body.

The composition provided by the present specification may include a combination of ferrous sulfate and lipoic acid.

The composition may include about 0.5 μM to 1 mM of ferrous sulfate and about 0.5 μM to 1 mM of lipoic acid.

In one embodiment, the composition may include about 0.5 μM to 100 μM of ferrous sulfate and about 0.5 μM of lipoic acid.

In another embodiment, the composition may include about 0.5 μM to 100 μM of ferrous sulfate and about 200 μM of lipoic acid.

In another embodiment, the composition may include about 0.5 μM to 100 μM of ferrous sulfate and about 400 μM of lipoic acid.

Function of the main ingredients of the composition

In addition to the above-described general function, the composition including at least one of ferrous sulfate and lipoic acid disclosed in the present specification may be a material that plays a major role in increasing cell activity.

Specifically, the lipoic acid may play a major role in increasing the activity of cells. By combining the ferrous sulfate and the lipoic acid, the effect of increasing the activity of the cells may be increased.

The term "cell activity" means

i) proliferation of cells,

ii) cell regeneration,

iii) anti-aging of existing cells or

iv) may include a cell whitening effect.

In addition to this, it may include the meaning of the general term "active" used in the art.

In one embodiment, the composition containing the lipoic acid may be a material that plays a major role in increasing the activity of cells.

In another embodiment, the composition including the combination of ferrous sulfate and lipoic acid may be a material that plays a major role in increasing the activity of cells.

Cells subject to the increase in activity may be skin cells, and the skin cells may include cells present in the epidermis and the dermis.

The skin cells may include one or more of keratinocytes, melanocytes, Langerhans cells, and Merkels cells.

The skin cells may include fibroblasts present in the dermis of the skin.

The skin cells may include one or more of a fully differentiated cell, a progenitor cell, and a stem cell.

The term "progenitor cell" is a cell in the middle of a stem cell and a fully differentiated cell, and can differentiate into a specific cell and, unlike stem cells, can divide only a limited number of times.

As one of the methods for improving the skin condition, it is necessary to increase the activity of the cells, because the activity of the cells decreases due to skin aging or damage.

For example, in the case of skin aging or damage, the activity of stem cells as well as completely differentiated cells among the skin cells decreases, and stem cells with reduced activity have the ability to produce self cells compared to stem cells that do not have decreased activity. This drops to about a third. As the number of own cells decreases, the skin's ability to regenerate decreases, and a typical example of this is wrinkles.

Therefore, there is a need for increased cellular activity for skin regeneration, anti-aging or maintaining skin elasticity.

Cells to be targeted for the increase in activity may not be limited to fully differentiated cells.

For example, cells to be subjected to increased activity may be muscle cells, neurons, germ cells, immune cells, or bone cells.

In addition, the degree of differentiation of cells to be subjected to the increase in activity is not limited.

For example, fully differentiated cells, progenitor cells, or stem cells may all be targets of increased activity.

Specifically, embryonic stem cells, adult stem cells, or induced pluripotent stem cells may be targeted for increased activity.

Optional components of the composition for cell activation

The composition for activating skin cells disclosed by the present application may include optional constituents other than ferrous sulfate and lipoic acid.

The optional component may include one or more of nutrients, ribonucleosides, deoxyribonucleosides, protein, animal-derived serum, carrier component, and other components that can be used for skin improvement and other substances. And is not limited thereto.

In addition, the optional components may be blended within a range that does not impair the objects and effects of the present invention.

In one embodiment, the nutrient may be one or more selected from the group consisting of amino acids, sugars, vitamins, inorganic salts, and lipids, but is not limited thereto.

For example, amino acids include Glycine, Arginine, Arginine hydrochloride, Glutamine, Histidine, Isoleucine, Serine, and Leucine. , Lysine, Methionine, Phenylalanine, Threonine, Tryptophan, Valine, Tyrosine, Cysteine, Cysteine HCL (L-Cysteine hydrochloride- H2O), Proline, L-Asparagine-H2O, Asparagine, Glutamic acid, Aspartic Acid, Histidine hydrochloride-H2O, And one or more may be selected from the group consisting of alanine.

For example, the saccharide is one or more selected from the group consisting of glucose, galactose, ribose, fructose, sucrose, lactose, and maltose. I can.

For example, the vitamin is a group consisting of vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin C, vitamin D, vitamin E, and vitamin K, or One or more may be selected from the group consisting of precursors.

Specifically, the vitamins are retinol, retinal, retinoid, carotenoid, thiamine, thiamine hydrochloride, riboflavin, niacin, niacinamide, pantothenic acid, pyridoxine, pyridoxine HCL, pyridoxine hydrochloride. Doxamine, pyridoxal, biotin, folic acid, folinic acid, cyanocobalamin (Vitamin B12), hydroxycobalamin, methylcobalamin, ascorbic acid, ergocalciferol, cholecalciferol, tocopherol , Tocotrienol, phylloquinone, menaquinone, calcium pantothenate (D-Calcium pantothenate), folic acid (Folic Acid), riboflavin and inositol (i-Inositol) one or more may be selected from the group consisting of, but is not limited thereto. .

For example, inorganic salts include cobalt (Co), copper (Cu), fluorine (F), iron (Fe), potassium (K), manganese (Mn), molybdenum (Mo), nickel (Ni), One or more may be selected from the group consisting of selenium (Se), silicon (Si), bismuth (Bi), vanadium (V), and zinc (Zn). Specifically, copper sulfate (CuSO ₄ ·5H ₂ O), sodium chloride (NaCl), calcium chloride (CaCl ₂ ·H ₂ O), potassium chloride (KCl), magnesium sulfate (MgSO ₄ ), sodium phosphate (NaH ₂ PO ₄ ·H ₂ O), sodium hydrogen carbonate (NaHCO ₃ ), magnesium chloride (MgCl ₂ ·6H ₂ O), iron nitrate (Fe(NO ₃ ) ₃ ·9H ₂ O) and zinc sulfate (ZnSO ₄ ·7H ₂ O) One or more may be selected from, but is not limited thereto.

For example, the lipid may be linoleic acid, but is not limited thereto.

In another embodiment, the protein may be a protein involved in the cellular signaling system or an extracellular matrix protein, and these may be substances secreted from cells.

For example, the ribonucleosides may be adenosine, cytidine, guanosine, or uridine.

For example, deoxyribonucleosides may be thymidine.

For example, the protein involved in the cellular signaling system may be a cytokine or a growth factor, but is not limited thereto.

Specifically, cytokines (Cytokine) are MCP-2, MCP-4, MDC, NAP-2, ICAM-1, MIP-1α, MIP-1β, sTNF-RII, sTNF-RI, TIMP-1, TIMP-2 , uPAR, CD14, CXCL-16, MMP-9, PDGF-AA, and TGFβ2 may be one or more selected from the group consisting of, but is not limited thereto.

Specifically, growth factors include insulin-like growth factor (IGF), epidermal growth factor (EGF), fibroblast growth fac-tor (FGF), and nerve growth. Factors (Nerve growth factor, NGF), Trans-forming growth factor (TGF), Platelet-derived growth fac-tor (PDGF), Bone-derived growth factor, BDF) and colony stimulation factor (CSF) may be selected from the group consisting of one or more, but is not limited thereto.

For example, the extracellular matrix protein may be one or more selected from the group consisting of collagen, elastin, hyaluronic acid, and fibronectin.

In another embodiment, the animal-derived serum may be bovine-derived serum, but is not limited thereto.

For example, the bovine serum may be one or more selected from the group consisting of Fetal Bovine Serum (FBS), Bovine Calf Serum (BCS), and Fetal Calf Serum (FCS).

The composition including at least one of ferrous sulfate and lipoic acid provided herein may not contain the animal-derived serum.

In another embodiment, the carrier component may be one or more of a medium composition for cell culture, a cosmetic carrier, and a pharmaceutical carrier, but is not limited thereto.

For example, the medium composition for cell culture may be selected from the group consisting of distilled water, PBS (Phosphate-buffered saline), physiological saline, basic medium, and modified medium.

Specifically, the basic medium is DMEM low glucose, DMEM high glucose, KGB-SFM, MEM, BME, RPMI 1640, F-10, F-12, α-MEM, G-MEM, IMDM, MacCoy's 5A, AmnioMax, AmnioMaxII One or more may be selected from the group consisting of complete Medium, DMEM-F12 and Chang's Medium, and MesenCult-XF Medium, but is not limited thereto.

Specifically, the modified medium may be a modified medium of DMEM, a modified medium of α-MEM, or a modified medium of F-10, but is not limited thereto.

For example, a composition that can be used as an ingredient of a cosmetic carrier may be composed of an ingredient permitted as a cosmetic ingredient notified by the Ministry of Food and Drug Safety.

Specifically, if the cosmetic formulation is a paste, gel, or cream, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc and zinc oxide as cosmetic carriers One or more may be selected from the group consisting of, but is not limited thereto.

Specifically, when the cosmetic formulation is a powder or spray, one or more may be selected from the group consisting of lactose, talc, silica, aluminum hydroxide, calcium silicate, and polyamide powder as a cosmetic carrier component, but is not limited thereto. . More specifically, the spray may additionally include a propellant. For example, the propellant may be one or more selected from the group consisting of chlorofluorohydrocarbon, propane, butane and dimethyl ether.

Specifically, when the cosmetic formulation is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent may be used as a cosmetic carrier component. More specifically, the group consisting of water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol and fatty acid ester of sorbitan One or more can be selected from.

Specifically, when the cosmetic formulation is a suspension, as a cosmetic carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, It may be one or more selected from the group consisting of microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tracanite.

Specifically, when the cosmetic formulation is a surfactant-containing cleansing, as cosmetic carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, Fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohols, fatty acid glycerides, fatty diethanolamides, vegetable oils, lanolin derivatives and ethoxylated glycerol fatty acid esters may be selected from the group consisting of one or more.

For example, a composition that can be used as a component of a pharmaceutical carrier may be composed of an ingredient permitted as a pharmaceutical ingredient notified by the KFDA.

Specifically, pharmaceutical carriers included in pharmaceuticals include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose. , Water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto.

In addition, a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. may be additionally included in addition to the above components.

More specifically, in the case of non-aqueous solvents or suspensions for parenteral administration, one from the group consisting of vegetable oils such as propylene glycol, polyethylene glycol, olive oil, and esters such as ethyloleate as pharmaceutical carrier components. The above may be selected, but is not limited thereto.

More specifically, in the case of suppositories for parenteral administration, one or more selected from the group consisting of Witepsol, Macrogol, Tween 61, Kakaoji, Laurinji, and glycerogelatin as pharmaceutical carrier components May be, but is not limited thereto.

More specifically, in the case of a solid preparation for oral administration, an excipient or a lubricant may be included as a pharmaceutical carrier component. More specifically, in the case of the excipient, one or more may be selected from the group consisting of starch, calcium carbonate, sucrose, lactose, and gelatin, but is not limited thereto. More specifically, the lubricant may be magnesium stearate or talc, but is not limited thereto.

More specifically, in the case of a liquid formulation for oral administration, water, liquid paraffin, or an excipient may be included as a pharmaceutical carrier component, but is not limited thereto. More specifically, the excipient may be a humectant, sweetener, fragrance or preservative.

In another embodiment, the component that can be used for skin improvement may be an antioxidant, a melanin pigment synthesis inhibitor, a toxic substance acting on cells containing melanin, or a substance that exfoliates dead skin cells, but is not limited thereto.

Specifically, the antioxidant may be carotenoids, flavonoids, isoflavones, vitamins or minerals, but is not limited thereto. More specifically, the carotenoids may be beta-carotene, lycopene or lutein. More specifically, the flavonoids may be anthocyanins, catechins, resveratrol or proanthocyanidins.

More specifically, the isoflavones may be genistein and daidzein. More specifically, the vitamin may be tocopherol.

Specifically, the melanin pigment synthesis inhibitory substance may be a tyrosinase inhibitor or a substance that removes active oxygen. More specifically, the tyrosinase inhibitor may be a mulberry extract, arbutin, licorice extract, vitamin C, kojic acid, or alpha-Bisabolol, but is not limited thereto.

More specifically, the substance that removes active oxygen may be an antioxidant substance.

Specifically, the toxic substance acting on cells containing melanin pigment may be hydroquinone or a derivative thereof.

Specifically, the substance that exfoliates the dead skin cells may be alpha hydroxy acid (AHA), but is not limited thereto.

In another embodiment, other cellular energy sources, antibiotics, flavors, preservatives, or pigments may be additionally included in the composition including at least one of ferrous sulfate and lipoic acid provided herein. And is not limited thereto. For example, the energy source of other cells may be sodium pyruvate. For example, the preservative may be phenoxyethanol or 1,2-hexanediol.

The composition disclosed herein may further include additional components in addition to the above-described components.

Effect of the composition for cell activation

By the present specification, various compositions that cause a cell activating effect can be provided.

In one embodiment, a cell activation effect may be exhibited by a composition containing lipoic acid as an essential component. Specifically, the cell activation effect may be exhibited by the lipoic acid (Lipoic aicd).

In another embodiment, a composition containing a combination of ferrous sulfate and lipoic acid as an essential component may exhibit a cell activation effect. Specifically, a cell activation effect may be exhibited by the combination of ferrous sulfate and lipoic acid.

The activity of skin cells may be increased by the composition provided herein, and the skin cells may be cells of the epidermis or the dermis.

In one embodiment, the activity of keratinocytes, melanocytes, Langerhans cells, or Merkels cells present in the epidermis of the skin may be increased.

In another embodiment, the activity of fibroblasts present in the dermis of the skin may be increased.

For example, the activity of stem cells present in the epidermis of the skin or the stem cells present in the dermis of the skin may be increased by the composition provided herein.

The activity of differentiated skin cells (fully differentiated cells) may be increased by the composition provided herein.

By the composition provided herein, the activity of skin stem cells or progenitor cells may be increased.

The composition may increase the activity of not only skin cells, but also muscle cells, nerve cells, germ cells, immune cells, or bone cells, and the degree of differentiation is not limited. For example, a fully differentiated cell, a progenitor cell, or a stem cell may have increased activity.

(1) Effect of the composition for cell activation -cell proliferation

The composition may increase the proliferation effect of cells.

According to some embodiments provided in the present application, when compared with the cell proliferation effect by the composition not containing ferrous sulfate and lipoic acid, the ferrous sulfate and/or lipoic acid ( The cell proliferation effect by the composition containing Lipoic acid) may be about twice or more high.

For example, when compared to the skin stem cell proliferation effect by a composition not containing ferrous sulfate and lipoic acid, the combination of ferrous sulfate and lipoic acid in a specific concentration Skin cell proliferation effect by the composition comprising as can be high.

Specifically, the skin cell proliferation effect of the composition containing ferrous sulfate in an amount of about 0.5 μM to about 500 μM and lipoic acid in an amount of about 0.5 μM to about 500 μM is determined by ferrous sulfate and lipoic acid (Ferrous sulfate) and lipoic acid. Lipoic acid) may be higher than the proliferation effect of a composition not containing.

For another example, the effect of proliferating skin cells by a composition containing ferrous sulfate and lipoic acid in a specific ratio may be high.

Specifically, the skin cell proliferation effect of the composition containing ferrous sulfate and lipoic acid in a molar ratio of 1:1 to 1:10 was obtained by using ferrous sulfate and lipoic acid. It may be higher than the skin cell proliferation effect by the composition not containing.

Depending on the type of skin cells, the molar ratio of ferrous sulfate and lipoic acid, which may increase the cell proliferation effect, may be different.

For example, in the case of skin cells containing a lot of fibroblasts, when ferrous sulfate and lipoic acid are contained in a molar ratio of 1:1 to 1:2, a high cell proliferation effect is achieved. Can appear.

As another example, in the case of skin cells containing a lot of skin stem cells, high cells when ferrous sulfate and lipoic acid are contained in a molar ratio of 1:4 to 1:8. There may be a proliferative effect.

According to some embodiments provided in the present application, the cell proliferation effect by the composition including the combination of ferrous sulfate and lipoic acid is only the ferrous sulfate without the lipoic acid. It may be about twice as high as the cell proliferation effect by the containing composition.

In addition, the cell proliferation effect by the composition containing the combination of ferrous sulfate and lipoic acid is the cell proliferation effect by the composition containing only the lipoic acid without the ferrous sulfate. Can be higher than

According to some embodiments provided in the present application, a specific proliferation effect of skin stem cells may be exhibited by a composition including ferrous sulfate and/or lipoic acid.

For example, compared to the proliferation effect of bone marrow-derived-mesenchymal stem cells or fat-derived-mesenchymal stem cells, proliferation of skin stem cells by a composition containing ferrous sulfate and/or lipoic acid The effect can be 1.5 to 3 times higher. Specifically, compared to adult stem cells other than skin stem cells, the proliferation effect of skin stem cells by the composition containing ferrous sulfate and/or lipoic acid may be 2 to 3 times higher. have.

For another example, the proliferation effect of skin stem cells by a composition containing ferrous sulfate and lipoic acid is obtained by the composition containing ferrous sulfate or lipoic acid, respectively. May be higher than the proliferative effect caused by.

For example, when compared to the skin stem cell proliferation effect by a composition not containing ferrous sulfate and lipoic acid, the combination of ferrous sulfate and lipoic acid in a specific concentration Skin stem cell proliferation effect by the composition comprising as can be high.

Specifically, the effect of skin stem cell proliferation by a composition containing about 0.5 μM to about 500 μM of ferrous sulfate and about 0.5 μM to about 500 μM of lipoic acid was determined by ferrous sulfate and lipoic acid. acid) may be higher than the proliferation effect of a composition not containing.

More specifically, the effect of skin stem cell proliferation by the composition containing about 0.5 μM to about 200 μM of ferrous sulfate and about 100 μM to about 500 μM of lipoic acid was determined by ferrous sulfate and lipoic acid. acid) may be about 2 to 4 times higher than the proliferation effect by the composition not containing.

More specifically, the effect of skin stem cell proliferation by a composition containing about 0.5 μM to about 10 μM of ferrous sulfate and about 100 μM to about 500 μM of lipoic acid was determined by ferrous sulfate and lipoic acid (Ferrous sulfate) and lipoic acid. Lipoic acid) may be about 2 to 3 times higher than the proliferation effect of the composition not containing.

For example, the effect of skin stem cell proliferation by a composition containing about 0.5 μM to about 10 μM of ferrous sulfate and about 100 μM to about 300 μM of lipoic acid is determined by ferrous sulfate and lipoic acid. ) It may be about 2 to 3 times higher than the proliferative effect by the composition not containing.

As another example, the effect of skin stem cell proliferation by a composition containing about 0.5 μM of ferrous sulfate and about 200 μM of lipoic acid is obtained by a composition that does not contain ferrous sulfate and lipoic acid. It can be about 2 to 3 times higher than the proliferative effect by.

As another example, the effect of skin stem cell proliferation by a composition containing about 0.5 μM to about 10 μM of ferrous sulfate and about 300 μM to about 500 μM of lipoic acid is determined by ferrous sulfate and lipoic acid. ) It may be about 2 to 3 times higher than the proliferative effect by the composition not containing.

As another example, the effect of skin stem cell proliferation by a composition containing about 0.5 μM of ferrous sulfate and about 400 μM of lipoic acid can be found in a composition that does not contain ferrous sulfate and lipoic acid. It can be about 2 to 3 times higher than the proliferative effect by.

More specifically, the effect of skin stem cell proliferation by the composition containing about 10 μM to about 60 μM of ferrous sulfate and about 100 μM to about 500 μM of lipoic acid was determined by ferrous sulfate and lipoic acid. acid) may be about 3 to 4 times higher than the proliferation effect by the composition not containing.

For example, the effect of skin stem cell proliferation by a composition containing about 10 μM to about 60 μM of ferrous sulfate and about 100 μM to about 300 μM of lipoic acid is determined by ferrous sulfate and lipoic acid. It may be about 3 to 4 times higher than the proliferative effect by the composition not containing.

As another example, the effect of skin stem cell proliferation by a composition containing about 50 μM of ferrous sulfate and about 200 μM of lipoic acid is obtained by a composition not containing ferrous sulfate and lipoic acid. It can be about 3 to 4 times higher than the proliferative effect.

As another example, the effect of skin stem cell proliferation by a composition containing about 10 μM to about 60 μM of ferrous sulfate and about 300 μM to about 500 μM of lipoic acid is determined by ferrous sulfate and lipoic acid. It may be about 3 to 4 times higher than the proliferative effect by the composition not containing.

As another example, the effect of skin stem cell proliferation by a composition containing about 50 μM of ferrous sulfate and about 400 μM of lipoic acid was obtained by a composition not containing ferrous sulfate and lipoic acid. It can be about 3 to 4 times higher than the proliferative effect.

More specifically, the effect of skin stem cell proliferation by a composition containing about 60 μM to about 200 μM of ferrous sulfate and about 100 μM to about 500 μM of lipoic acid was determined by ferrous sulfate and lipoic acid. acid) may be about 3 to 4 times higher than the proliferation effect by the composition not containing.

For example, the effect of skin stem cell proliferation by a composition containing about 60 μM to about 200 μM of ferrous sulfate and about 100 μM to about 300 μM of lipoic acid is determined by ferrous sulfate and lipoic acid. It may be about 3 to 4 times higher than the proliferative effect by the composition not containing.

As another example, the effect of skin stem cell proliferation by a composition containing about 100 μM of ferrous sulfate and about 200 μM of lipoic acid is obtained by a composition not containing ferrous sulfate and lipoic acid. It can be about 3 to 4 times higher than the proliferative effect.

As another example, the effect of skin stem cell proliferation by a composition containing about 60 μM to about 200 μM of ferrous sulfate and about 300 μM to about 500 μM of lipoic acid is determined by ferrous sulfate and lipoic acid. It may be about 3 to 4 times higher than the proliferative effect by the composition not containing.

As another example, the effect of skin stem cell proliferation by a composition containing about 100 μM of ferrous sulfate and about 400 μM of lipoic acid is obtained by a composition not containing ferrous sulfate and lipoic acid. It can be about 3 to 4 times higher than the proliferative effect.

For another example, the effect of proliferating skin stem cells by a composition containing ferrous sulfate and lipoic acid in a specific ratio may be high.

Specifically, the skin stem cell proliferation effect by the composition containing ferrous sulfate and lipoic acid in a molar ratio of 1:1 to 1:10 was obtained by ferrous sulfate and lipoic acid. It may be higher than the skin stem cell proliferation effect by the composition not containing.

More specifically, the skin stem cell proliferation effect of the composition containing ferrous sulfate and lipoic acid in a molar ratio of 1:2 to 1:4 was determined by ferrous sulfate and lipoic acid. ) It may be about 3 to about 4 times higher than the skin stem cell proliferation effect by the composition not containing.

More specifically, the effect of proliferating skin stem cells by the composition containing ferrous sulfate and lipoic acid in a molar ratio of 1:4 to 1:8 is ferrous sulfate. And it may be about 3 to about 4 times higher than the proliferation effect of skin stem cells by a composition not containing lipoic acid.

For example, the effect of skin stem cell proliferation by a composition containing ferrous sulfate and lipoic acid at a molar ratio of 1:4 does not include ferrous sulfate and lipoic acid. It can be about 3 to 4 times higher than the proliferative effect by the composition.

More specifically, the skin stem cell proliferation effect of the composition containing ferrous sulfate and lipoic acid in a molar ratio of 1:8 to 1:10 was determined by ferrous sulfate and lipoic acid. ) It may be about 3 to about 4 times higher than the skin stem cell proliferation effect by the composition not containing.

As another example, the effect of skin stem cell proliferation by a composition containing ferrous sulfate and lipoic acid at a molar ratio of 1:8 is not containing ferrous sulfate and lipoic acid. It can be about 3 to 4 times higher than the proliferative effect by the composition.

As another example, the effect of skin stem cell proliferation by a composition containing a combination of ferrous sulfate and lipoic acid is a composition containing only ferrous sulfate without the lipoic acid. It can be about 4 times higher than that of the skin stem cell proliferation effect.

In addition, the cell proliferation effect by the composition containing the combination of ferrous sulfate and lipoic acid is the skin stem cells by the composition containing only the lipoic acid without the ferrous sulfate. It can be about 1.5 to about 4 times higher than the proliferative effect.

As another example, the skin stem cell proliferation effect by the composition containing the combination of ferrous sulfate and lipoic acid may be about 1.5 to about 2 times higher than that of the primary fibroblast proliferation effect. I can.

According to some examples provided in the present application, even when the compositions provided by some examples of the present application do not contain animal-derived serum, a cell proliferation effect as high as that of treatment with animal-derived serum may be exhibited.

Specifically, when the composition is treated with fibroblasts, proliferation may be promoted as much as when the animal-derived serum is treated.

Specifically, when the composition is treated with skin stem cells, proliferation can be promoted as much as when the animal-derived serum is treated.

Specifically, when the composition is treated with skin stem cells, proliferation may be promoted more than when the animal-derived serum is treated.

More specifically, the animal-derived serum may be bovine-derived serum, but is not limited thereto. Specifically, the proprietary serum may be Fetal Bovine Serum (FBS), Bovine Calf Serum (BCS), or Fetal Calf Serum (FCS). More specifically, the animal-derived serum may be 10% FBS.

(2) Effect of the composition for cell activation-Cell regeneration

The regeneration effect of stem cells or progenitor cells may be exhibited by the composition including ferrous sulfate and/or lipoic acid.

In one embodiment, the cell regeneration effect by the composition containing ferrous sulfate and lipoic acid is a cell by a composition not containing ferrous sulfate and lipoic acid. May be higher than the regeneration effect.

In one embodiment, the cell regeneration effect by the composition comprising the ferrous sulfate and lipoic acid in a molar ratio of 1:2 to 1:8 is the ferrous sulfate and the lipoic acid ( Lipoic acid) may be 2 to 3 times higher than the cell regeneration effect by the composition not containing.

In another embodiment, the cell regeneration effect by the composition containing ferrous sulfate and lipoic acid is the regeneration effect by the composition containing ferrous sulfate or the lipoic acid, respectively. Can be higher.

For example, the effect of cell regeneration by a composition containing a combination of ferrous sulfate and lipoic acid is a cell regeneration effect by a composition containing only ferrous sulfate without the lipoic acid (Lipoic acid). It can be about 2 to about 4 times higher than the effect.

As another example, the cell regeneration effect of the composition containing a combination of ferrous sulfate and lipoic acid is due to the composition containing only the lipoic acid without the ferrous sulfate. It can be about 2 to about 4 times higher than the cell regeneration effect.

(3) Effects of the composition for cell activation-Prevention of cell aging

Cells may be prevented from aging by the composition, and cells may be prevented from aging by inhibiting expression of senescence regulators, but the present invention is not limited thereto.

The term “aging modulator” is a substance that inhibits the action of an enzyme that regulates the cell cycle, and as the expression of the aging regulator is inhibited, aging may be prevented.

In one embodiment, the expression of aging modulators may be suppressed by the composition provided herein.

For example, the aging regulator may be of the Cyclin-dependent Kinase Inhibitors (CKIs) family.

Specifically, CKIs may be CIP/KID series or INK series.

More specifically, the CIP/KID series may be P21, P27, or P57.

More specifically, the INK series may be P16, P15, P18 or P19.

For example, the aging modulator can be P53 or p-P53.

(4) Effect of the composition for cell activation -cell whitening

The cell whitening effect may be exhibited by the composition containing ferrous sulfate and/or lipoic acid.

The whitening effect may be an effect by inhibiting the synthesis of melanin pigments in cells, decomposing melanin pigments generated in cells, destruction of cells including melanin pigments, or exfoliation of dead skin cells.

For example, the composition provided herein may be a substance that acts as a tyrosinase inhibitor.

For example, the composition provided herein may be a material that serves to scavenge active oxygen or free radicals. For example, the composition provided herein may be a substance that serves to destroy cells including melanin pigment. For example, the composition provided herein may be a material that serves to exfoliate dead skin cells.

Specifically, the composition provided in the present specification may exhibit a whitening effect higher than that of arbutin, niacinamide, kojic acid and/or RSV, which are well known as substances exhibiting a whitening effect. have.

Specifically, the cell whitening effect of the composition containing ferrous sulfate and lipoic acid provided in the present specification is not limited to the composition containing only the lipoic acid without the ferrous sulfate. Can be high compared to the cell whitening effect caused by.

Specifically, the cell whitening effect of the composition including ferrous sulfate and lipoic acid provided in the present specification is not limited to the composition containing only ferrous sulfate without the lipoic acid. Can be high compared to the cell whitening effect caused by.

(5) Effect of composition for cell activation -Improvement of skin condition

By the composition provided herein, the skin condition improvement effect may be exhibited, and the condition of the epidermis, dermis, or subcutaneous fat layer may be improved.

The skin condition improvement effect may appear not only through cell activation but also through inhibition of specific substances, but is not limited thereto.

In one embodiment, the composition provided herein may exhibit a wound healing effect, which may be an effect exhibited by proliferation of skin cells and cell regeneration.

For example, a proliferative effect of epidermal cells may be exhibited by the composition provided herein, thereby regenerating damaged epidermal cells and forming a skin barrier. Specifically, the epidermal cells may be keratinocytes, progenitor cells or stem cells thereof.

For another example, the composition provided herein may produce extracellular matrix components of the dermis or proliferate dermal cells, thereby improving elasticity and wrinkles. Specifically, the extracellular matrix component may be collagen, elastin or hyaluronic acid. Specifically, the dermal cells may be fibroblasts, progenitor cells or stem cells thereof.

In another embodiment, a skin whitening effect may be exhibited by the composition provided herein.

For example, the synthesis of melanin pigments in cells may be inhibited by the composition provided herein, and thus, a skin whitening effect may be exhibited.

For another example, melanin pigment generated in cells may be decomposed by the composition provided herein, and thus, a skin whitening effect may be exhibited.

Cosmetic composition for cell activation

The composition provided herein may be a cosmetic composition for cell activation.

The cosmetic composition may be composed of ingredients permitted as cosmetic raw materials notified by the Ministry of Food and Drug Safety.

The cosmetic composition for cell activation includes at least one of ferrous sulfate and lipoic acid.

The cosmetic composition may be formulated as a basic cosmetic, makeup cosmetic, body cosmetic, or hair cosmetic, but is not limited thereto.

For example, basic cosmetics include softening lotion, nutritional lotion, lotion, skin, body lotion, nutrition cream, massage cream, moisture cream, sun cream, hand cream, eye cream, essence, cleansing foam, cleansing cream, cleansing water, pack, It may be formulated as a gel, oil or patch, but is not limited thereto.

For example, the makeup cosmetic may be formulated as a lipstick, a makeup base, a powder or a foundation, but is not limited thereto.

For example, the cosmetic for the body may be formulated as a body cleanser, toothpaste, or mouthwash, but is not limited thereto.

For example, the cosmetic composition for hair may be formulated with a hairdressing agent such as shampoo, conditioner, hair tonic, gel, mousse, hair essence, and hair pack, a wool or hair dye, but is not limited thereto.

The cosmetic composition may contain a medium for cell culture, but may contain only ingredients permitted as raw materials for cosmetics.

In one embodiment, the medium for cell culture may contain one or more of ferrous sulfate and lipoic acid, and in general, components that may be added during cell culture may be additionally included.

The cosmetic composition may include a cell culture solution.

The term "cell culture medium" refers to a material obtained by purification from a medium containing a substance secreted from cells through cell culture or a medium containing a substance secreted from cells through cell culture.

The material secreted from the cell through the cell culture may be a peptide (Peptide).

In one embodiment, the peptide may be a cytokine.

For example, cytokines are MCP-2, MCP-4, MDC, NAP-2, ICAM-1, MIP-1α, MIP-1β, sTNF-RII, sTNF-RI, TIMP-1, TIMP-2, uPAR , CD14, CXCL-16, MMP-9, PDGF-AA or TGFβ2, but is not limited thereto.

In another embodiment, the peptide may be a growth factor.

For example, growth factor is insulin-like growth factor (IGF), epidermal growth factor (EGF), fibroblast growth fac-tor (FGF), nerve Nerve growth factor (NGF), platelet-derived growth factor (PDGF), bone-derived growth factor (BDF) or colony stimulation factor (CSF) And is not limited thereto.

A cell activation effect may be exhibited by the cosmetic composition, for example, an activation effect of keratinocytes or fibroblasts may be exhibited.

Specifically, the effect of activating epidermal or dermal stem cells may be exhibited.

Ultimately, anti-aging effect, regeneration effect, wrinkle improvement effect, whitening effect, and skin elasticity effect may be exhibited by the cosmetic composition, but is not limited thereto.

The cosmetic composition may be prepared by a method that is obvious to those of ordinary skill in the art.

The cosmetic composition may be prepared through an emulsification, solubilization, or dispersion process, but is not limited thereto.

In one embodiment, the emulsification is a high temperature emulsification method (Continental Method), an inversion emulsification method (Inversion Emulsification Method), a D phase emulsification method (SSA phase Emulsification Method), a PIT Emulsification Method, or a liquid crystal emulsification method. It may be based on the LC Emulsification Method, but is not limited thereto.

In another embodiment, solubilization may be diluted by adding an aqueous phase or oil phase to a mixture of a surfactant and a substance to be solubilized. As another example, solubilization may be by dissolving a surfactant in an aqueous or oily phase and adding a substance to be solubilized.

In another embodiment, dispersion may be mixing and dissolving by rotating a stirrer into a dispersion medium.

Pharmaceutical composition for cell activation

The composition provided herein may be a pharmaceutical composition for cell activation.

The pharmaceutical composition may be composed of ingredients permitted as pharmaceutical ingredients notified by the KFDA.

In the case of a pharmaceutical composition, it can be prepared by a conventional method used by those skilled in the art.

The pharmaceutical composition may be an internal solid, an injection, an eye drop, an internal solution, an external solution, an ointment, a patch, a solid or a liquid, but is not limited thereto.

For example, the solid contents are tablets, vaginal tablets, capsules, powders, granules, pills, troches, syrups, inhalants, oral disintegrating tablets, chewable tablets, effervescent tablets, dispersing tablets, dissolving tablets, foaming granules, multi-drugs, oral It may be a soluble tablet, a sublingual tablet, a bakkal tablet, an adhesive tablet, a gum agent, an oral dissolving film, an inhalation powder, a nasal powder, an extract or an oral jelly.

For example, the injection may be an injection, a powder injection, an infusion solution, a lyophilized injection, a transplant, a continuous injection, a peritoneal dialysis agent, a perfusion agent, or a dialysis agent.

For example, the liquid contents are oral solutions, syrups, emulsions, suspensions, elixirs, lemonades, tinctures, liquid extracts, alcohols, fragrances, presumptions, immersions, inhalants, inhalants, extracts, oral It may be a spray, inhalation aerosol, nasal drops, gargle, or oral jelly.

For example, the external solution may be a lotion, a liniment, an aerosol, an external aerosol, a pump spray, an enema, a gargle, a hemodialysis agent, a dialysis agent, an ear drop, an ear drop, or a nasal drops.

For example, the ointment may be a cream, a paste, a liniment, an ointment, a suppository, a gel, an oral semi-solid, a rectal semi-solid, a vaginal suppository, an enema, or an ear drop.

For example, the patch may be a cataplasmase or a transdermal absorber.

For example, the solid agent may be an external powder, an inhalant, an inhaled powder, an external solid, a gargle, a hemodialysis agent, a dialysis agent, an ear drop, or a nasal powder.

Media composition for cell activation

The composition provided herein may be a medium composition for cell culture for cell activation.

The medium for cell culture may be a medium for culturing fully differentiated cells, a medium for culturing progenitor cells, or a medium for culturing stem cells.

For example, a fully differentiated cell may be a muscle cell, a nerve cell, a skin cell, a germ cell, an immune cell, or a bone cell, but is not limited thereto.

Specifically, the skin cells may be epidermal cells of the skin or dermal cells of the skin.

More specifically, the epidermal cells of the skin may be keratinocytes or melanocytes.

More specifically, the dermal cells of the skin may be fibroblasts.

For example, the stem cells may be embryonic stem cells, adult stem cells, or induced pluripotent stem cells.

Specifically, the adult stem cells may be neural stem cells, hematopoietic stem cells, mesenchymal stem cells, and skin stem cells, but are not limited thereto.

More specifically, the mesenchymal stem cells may be adipose-derived-mesenchymal stem cells, bone marrow-derived-mesenchymal stem cells, or cord blood-derived-stem cells, but are not limited thereto.

As described above, a cell activation effect may be exhibited by a composition including at least one of ferrous sulfate and lipoic acid provided in the present specification as an essential component.

In addition, those skilled in the art may further include other components in addition to ferrous sulfate and lipoic acid, which may be essential components.

The composition comprising at least one of ferrous sulfate and lipoic acid provided herein as an essential component may be a cosmetic composition, a pharmaceutical composition, or a medium for cell culture, but is not limited thereto.

A cosmetic composition, a pharmaceutical composition, or a cell culture medium containing at least one of ferrous sulfate and lipoic acid as an essential component may also exhibit a cell activation effect.

Hereinafter, the composition provided by the present specification and the effect of the composition will be described in more detail through examples.

[Example 1] Preparation of a medium composition containing at least one of ferrous sulfate and alpha-lipoic acid

example The inventors prepared the composition provided by the present specification by including ferrous sulfate and lipoic acid in various amounts in a modified medium in which some of the constituents of a commercial medium were changed.

The modified medium contains the constituents described in the following [Table 1].

The inventors of the present invention manufactured and used the modified medium by Weljin Co., Ltd. In the examples disclosed herein, the content of ferrous sulfate and lipoic acid in the medium was expressed as a molar concentration (mol/L; M).

Division Components Standard name mg/L CAS No. Amino Acids Glycine Glycine 50 56-40-6 L-Arginine Arginine 105 74-79-3 (L-) L-Glutamine Glutamine 292 56-85-9 (L-) L-Glutamic Acid Glutamic Acid 75 56-86-0 (L-) L-Asparagine-H2O Asparagine 50 5794-13-8 (monohydrate) L-Alanine Alanine 25 56-41-7 (L-) L-Aspartic acid Aspartic Acid 30 56-84-8 (L-) L-Cysteine hydrochloride-H2O Cysteine HCL 100 7048-04-06,52-89-1 L-Histidine Histidine 31 71-00-1 (L-), 4998-57-6 L-Isoleucine Isoleucine/Isoleucine 52.4 73-32-5 (L-) L-Leucine Leucine 52 61-90-5 (L-) L-Lysine Lysine 73 56-87-1 (L-) L-Methionine Methionine/Methionine 15 63-68-3 L-Phenylalanine Phenylalanine 32 62056-68-2 (L-) L-Serine Serine 25 56-45-1 (L-) L-Threonine Threonine 48 72-19-5 (L-) L-Tryptophan Tryptophan 10 73-22-3 (L-) L-Proline Proline 40 147-85-3 (L-) L-Valine Valine 46 72-18-4 (L-) Vitamins D-Calcium pantothenate Calcium pantothenate One 137-08-6 (D-) Folic Acid Polic Acid One 59-30-3 Niacinamide Niacinamide One 98-92-0 Pyridoxal hydrochloride Pyridoxine HCL One 58-56-0 (Pyridoxine HCl) Riboflavin Riboflavin 0.1 83-88-5 Thiamine hydrochloride Thiamine HCL One 67-03-08 (Thiamine HCl) Vitamin B12 Cyanocobalamin 1.36 68-19-9 (Cyanocobalamin) Ascorbic Acid Ascor Big Acid 50 50-81-7(L-) Biotin Biotin 0.1 58-85-5 i-Inositol Inositol 2 87-89-8 (myo-inositol/inositol), 643-12-9 (D-(+)-chiro-Inositol) Inorganic Salts Calcium Chloride (CaCl2) (anhyd.) Calcium chloride 200 10035-04-8 Magnesium Sulfate (MgSO4) (anhyd.) Magnesium sulfate 97.67 18939-43-0 Potassium Chloride (KCl) Potassium chloride 400 7447-40-7 Sodium Bicarbonate (NaHCO3) Sodium bicarbonate 2200 144-55-8 Sodium Chloride (NaCl) Sodium chloride 6800 7647-14-5 Sodium Phosphate monobasic (NaH2PO4-H2O) Sodium phosphate 140 7558-80-7 (Sodium Phosphate) Other Components D-Glucose (Dextrose) Glucose 1000 50-99-7 (D-) Sodium Pyruvate Sodium pyruvate 110 113-24-6 Ribonucleosides Adenosine Adenosine 10 58-61-8 Guanosine Guanosine 10 118-00-3

1-1: Preparation of a composition containing ferrous sulfate and α-Lipoic acid

The present inventors produced a composition Mix II in which ferrous sulfate and alpha-lipoic acid were included in the modified medium, and compositions Mix I and Mix III in which other components were included in the modified medium.

The ferrous sulfate and alpha lipoic acid were purchased from Sigma Aldrich.

In the following [Table 2], components excluding the modified medium component among the components of Mix I, Mix II and Mix III are disclosed.

Product label Low conc. High conc. MixⅠ D-pantothenic acid Ca-salt Vit.B5 5uM 1mM L-Ascorbic Acid Vit.C 0.3mM 1mM Pyridoxine Hydrochloride Vit.B6 5uM 1mM MixⅡ DL-6,8 Thioctic Acid α-Lipoic 0.5uM 500uM Ferrous Sulfate FeSO4 2uM 50uM MixⅢ Sodium Metasilicate Nonahydrate Na2O3 0.3uM 10uM Mangan(2) sulfat-Monohydrat (MnSO4 x H2O) MnSO4 0.001uM 1uM Kupfer(2) sulfat-Pentahydrat (CuSO4 5H2O) CuSO4 0.005uM 0.5uM Stannous Chloride dihydrate (SnCl2 · 2H2O) SnCl2 0.0005uM 100uM Ammonium heptamolybdate tetrahydrat( (NH4)6Mo7O24 4H2O) NH4 0.02uM 0.1uM

1-2: Preparation of a composition containing ferrous sulfate and α-Lipoic acid (hereinafter, Experimental Examples (a) to Experimental Examples (i))

In addition, the present inventors prepared compositions (Experimental Examples (a) to Experimental Examples (i)) by varying the content of ferrous sulfate and α-Lipoic acid in the modified medium for detailed concentration evaluation. Was produced.

In Table 3 below, the molar concentrations of ferrous sulfate and α-Lipoic acid included in each composition (Experimental Examples (a) to Experimental Examples (i)) are disclosed.

α-Lipoic FeSo4 Label 0.5uM 0.5uM Experimental Example (a) 200uM Experimental Example (b) 400uM Experimental Example (c) 200uM 0.5uM Experimental Example (d) 200uM Experimental Example (e) 400uM Experimental Example (f) 400uM 0.5uM Experimental Example (g) 200uM Experimental Example (h) 400uM Experimental Example (i)

1-3: Preparation of a composition containing ferrous sulfate and α-Lipoic acid (Experimental Examples (1) to Experimental Examples (9))

The present inventors produced 9 compositions (Experimental Example (1) to Experimental Example (9)) for treating Holoclone having stem cell properties, and the 9 compositions were low concentration of ferrous sulfate (Ferrous Sulfate). ).

In Table 4 below, the molar concentrations of ferrous sulfate and α-Lipoic acid included in each composition (Experimental Examples (1) to (9)) are disclosed.

α-Lipoic FeSO4 Label 0.5uM 0.5uM Experimental Example (1) 50uM Experimental Example (2) 100uM Experimental Example (3) 200uM 0.5uM Experimental Example (4) 50uM Experimental Example (5) 100uM Experimental Example (6) 400uM 0.5uM Experimental Example (7) 50uM Experimental Example (8) 100uM Experimental Example (9)

[Example 2] Effect of Fibroblast Proliferation by Treatment of a Composition Containing a Combination of Ferrous Sulfate and Alpha Lipoic Acid

The present inventors confirmed the cell proliferation effect of fibroblasts by treatment of several compositions provided by the present specification through CCK-8 assay (CCK8-Assay), which is common to those skilled in the art.

2-1: Effect of MixII treatment on fibroblast proliferation

The present inventors seeded human fibroblasts (Human Primary Fibroblast) in a 96 well plate equally at 4000 cells/well and cultured in a culture medium containing DMEM+10% FBS and 1% penicillin streptomycin for 1 day.

After washing the cultured human primary fibroblasts twice with PBS, 100 ul of Mix I, Mix II and Mix III were equally distributed to wells and cultured for 72 hours, and the proliferation effect was confirmed. The substances constituting Mix I, Mix II and Mix III were diluted at 1M or 0.5M stock concentration to suit the Low or High concentration, respectively, and treated on the human primary fibroblasts.

The proliferation effect was measured using the CCK-8 assay. After diluting 1/10 of the CCK-8 solution in DMEM medium without FBS, dispensing 100ul into each well, reacting in a 37°C incubator for 1-4 hours, and then 450nm It was measured in absorbance.

The present inventors have a proliferation effect of about twice or more in the MixII-High condition than in the condition where ferrous sulfate and α-Lipoic acid are not included (condition 0 in FIG. 1 or conditions of Mix I and Mix III). It was confirmed that appears. (See FIG. 1)

2-2: Confirmation of the degree of human fibroblast proliferation by a composition containing ferrus sulfate or a composition containing α-Lipoic acid

In the same manner as in the above-described Mix I, Mix II and Mix III treatment methods, the present inventors have demonstrated the effect of cell proliferation by a composition containing ferrous sulfate in a modified medium and a composition containing α-Lipoic acid in a modified medium. Confirmed.

As shown in Figure 2 (a), the present inventors have a cell proliferation effect of the composition containing the alpha-lipoic acid (α-Lipoic acid) in the modified medium is treated with the modified medium + 10% FBS (Figure 2 ( It was confirmed that the PC condition of a) was not as high as when it was the case of (hereinafter, positive control).

In addition, the present inventors, as shown in Figure 2 (b), when the cell proliferation effect by the composition containing the ferrous sulfate in the modified medium is also a positive control (PC condition of Figure 2 (b)) It was not as high as that, and it was confirmed that it was similar to the case where only the modified medium was treated (condition 0 in FIG. 2 (b)).

2-3: Effects of human fibroblast proliferation according to Experimental Examples (a) to (i)

The present inventors confirmed the cell proliferation effect by treating Experimental Examples (a) to (i) on the Human Primary Fibroblasts, respectively, in the same manner as the MixI, MixII, and MixIII treatment methods described above.

As shown in FIG. 3, when the human primary fibroblasts were treated with the experimental examples (b) to (i), only the modified medium was treated (condition 0 in FIG. 3; hereinafter, negative control) It was confirmed that a higher cell proliferation effect appeared.

Moreover, when the Experimental Example (e), Experimental Example (f), Experimental Example (h), and Experimental Example (i) were treated on the human primary fibroblast, the cells proliferated about 1.7 times more than the negative control. It was confirmed that the effect appeared.

The present inventors, than the cell proliferation effect of the composition containing the above-described alpha-lipoic acid (α-Lipoic acid) in a modified medium, a composition comprising a combination of the ferrous sulfate and alpha-lipoic acid (α-Lipoic acid) It was confirmed that the cell proliferation effect (refer to Figs. 2(a) and 3) is high.

In addition, cell proliferation by a composition comprising a combination of ferrous sulfate and alpha-lipoic acid, rather than the cell proliferation effect by the composition containing the above-described ferrous sulfate in a modified medium. It was confirmed that the effect (refer to FIGS. 2(b) and 3) was high.

In order to confirm whether the proliferation effect is due to skin stem cell proliferation, the present inventors experiment with a composition containing the ferrous sulfate and alpha-lipoic acid (Table 4). Examples (1) to (9)) were treated with holoclone isolated from human primary fibroblasts.

[Example 3] Effect of Proliferation of Skin Stem Cells by Treatment of a Composition Containing Ferrous Sulfate and Alpha Lipoic Acid

3-1: Obtaining skin stem cells

(1) Holoclone isolation from human dermal-derived fibroblasts

Holoclone is known to those skilled in the art as a cell having the properties of a stem cell (Nature Medicine volume 12, pages 1397-1402 (2006)).

Therefore, the present inventors conducted an experiment to find out the proliferation effect of holoclone in order to confirm that the proliferation of skin stem cells appears in the composition provided in the present application.

The present inventors isolated holoclones from human primary fibroblasts (Donor 12 years, Fore skin, passage 3), which were sold in the Department of Dermatology, Chung-Ang University through the following method.

The present inventors seeded so that 0.5 cells per well (1~2 or not/well) per well in a 96 well plate and cultured for 8 days without changing the culture medium, and DMEM+10% FBS was used as a culture medium. . The present inventors observed wells in which cell clusters were formed after 8 days.

The present inventors treated 0.25M Trypsin-EDTA for each well in which the cell cluster was formed, transferred to a 24-well plate for each cell lot, and then proliferated cells for 4 to 6 days, and only cells with rapid proliferation were selected. .

(2) Confirmation of skin stem cell marker

The present inventors derived dermal stem cell markers (CD29(+), CD34(-), CD44(+), CD90(+), CD105(+)) from the selected cells (Holoclone). Was confirmed. (See FIGS. 4 and 5). Through these results, it was confirmed that the holoclone isolated from human primary fibroblasts has the properties of skin stem cells.

The inventors of the present invention confirmed the proliferation effect by culturing the Holoclone in 25T Flask and treating several compositions provided by the present specification.

3-2: Confirmation of the degree of skin stem cell proliferation by a composition containing ferrus sulfate and a composition containing α-Lipoic acid

The present inventors seeded the same holoclone (holoclone) isolated from human primary fibroblasts (Donor 12 years, Fore skin, passage 3) in a 96 well plate at 4000 cells/well and DMEM+10% FBS , Incubated for 1 day in a culture medium containing 1% penicillin streptomycin.

After washing the cultured Holoclone twice with PBS, 100ul of a composition containing ferrous sulfate in a modified medium and a composition containing α-Lipoic acid in a modified medium were added to a well. After dispensing equally and culturing for 72 hours, the proliferation effect was confirmed.

The proliferative effect of the Holoclone can be determined through CCK-8 analysis (CCK8-Assay) in general for those skilled in the art. Confirmed.

When compared with the case where 10% FBS+ modified medium (hereinafter, positive control) (FBS condition in Fig. 6) was treated, when the composition containing only ferrous sulfate in the modified medium was treated, Holoclone There was no proliferative effect (see Fig. 6).

In addition, even when the composition containing only alpha-lipoic acid (α-Lipoic acid) was treated with the modified medium, the cell proliferation effect of Holoclone did not appear as much as the positive control (FBS condition in FIG. 7). (See Fig. 7)

3-3: Measurement of skin stem cell proliferation effect according to Experimental Examples (1) to (9)

(One) Holoclone proliferation effect confirmed

The present inventors seeded the same holoclone (Holoclone) isolated from human primary fibroblasts (Donor 12 years, Fore skin, passage 3) in a 96 well plate at 4000 cells/well and DMEM+10% FBS , Incubated for 1 day in a culture medium containing 1% penicillin streptomycin.

After washing the cultured Holoclone twice with PBS, 100ul of the compositions of Experimental Examples (1) to (9) were equally dispensed into wells and cultured for 72 hours, and then proliferative effect was observed. Confirmed.

The present inventors confirmed the proliferation effect by dispensing 100 μl of only the modified medium (condition 0 in FIG. 8) not containing ferrous sulfate and alpha-lipoic acid as a negative control to the well. In addition, the present inventors confirmed the proliferation effect by dispensing 100 μl of 10% FBS + modified medium (FBS condition of FIG. 8) into wells as a positive control.

The present inventors found that the proliferation effect of Holoclone according to Experimental Example (4) and Experimental Example (7) was 2.5 times higher than that of the negative control group, and Experimental Example (5), Experimental Example (6), Experimental Example It was confirmed that the proliferation effect of Holoclone according to (8) and Experimental Example (9) was about 3.5 times higher than that of the negative control group (see Fig. 8).

Moreover, it was confirmed that the proliferation effect of Holoclone according to Experimental Example (5), Experimental Example (6), Experimental Example (8) and Experimental Example (9) was similar to or higher than that of the positive control group. I did.

(2) Comparison of the proliferation effect of Holoclone and human primary fibroblast

The present inventors confirmed through the CCK-8 assay (CCK-8 Assay) that the proliferation effect of holoclones is greater than that of the human primary fibroblast proliferation effect by the composition provided by the present invention.

When the experimental example (8) was treated with human primary fibroblasts, the proliferation effect was 1.88 times higher than that of the case where only the modified medium was treated (negative control).

In comparison, when the experimental example (8) was treated with a holoclone having skin stem cell properties, the proliferation effect was 2.99 times higher than that of the negative control group. (See Fig. 9)

The present inventors confirmed that the proliferation effect of human primary fibroblasts was also shown by the experimental example (8), but the proliferation effect of holoclones was about twice as high.

3-4: Whether adult stem cells are proliferated from various sources

The present inventors have confirmed that the proliferation effect of skin stem cells is exhibited by the composition provided by the present application in light of the proliferative effect of the aforementioned Holoclone.

In order to confirm that the composition specifically affects skin stem cells, the present inventors conducted an experiment of treating the composition on bone marrow-derived stem cells and fat-derived stem cells.

Bone marrow-derived mesenchymal stem cells (Donor: Male 65 years) and adipose-derived mesenchymal stem cells (Donor: Male 47 years) used as adult stem cells were all purchased from Promo cells and used.

The present inventors treated the above Experimental Examples (1) to (9) to culture the bone marrow-derived stem cells and the adipose-derived stem cells. In addition, the present inventors treated only the modified medium (O in FIG. 10, 0 in FIG. 11) as a negative control on the bone marrow-derived stem cells and adipose-derived stem cells.

The present inventors seeded the bone marrow-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells in a 96 well plate equally at 4000 cells/well and cultured in a culture medium containing MEMα medium + 10% FBS and 0.5% Gentamicin for 1 day. After washing the cultured stem cells twice with PBS, Experimental Examples (1) to (9) containing ferrous sulfate and lipoic acid were equally dispensed into wells for 72 hours. After culturing during, the proliferation effect of the bone marrow-derived mesenchymal stem cells and the adipose-derived mesenchymal stem cells was confirmed.

As a result of analyzing the proliferation effect through CCK-8 analysis (CCK8-Assay), the proliferative effect of the bone marrow-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells was similar to or lower than that of the negative control group (FIGS. 10 and 11). Reference)

[Example 4] Checking the wound healing effect by a composition containing ferrous sulfate and alpha-lipoic acid

The present inventors conducted the following experiment to confirm whether the Wound Healing effect by Mix II containing ferrous sulfate and α-Lipoic acid appeared. As a comparative group, the Mix I composition and MixIII composition, the modified medium (negative control) and 10% FBS + the modified medium (positive control) were used.

The present inventors applied a scratch to human primary fibroblasts and treated the composition and confirmed the wound healing effect after 48 hours had elapsed.

N.Cont in FIG. 12 represents the negative control, and P.Cont in FIG. 12 represents the positive control. According to FIG. 12, in the case of treatment with MixII-low, the wound healing effect was shown as much as the conditions of the positive control group.

[Example 5] Confirmation of inhibition of cell senescence marker expression by a composition containing ferrous sulfate and alpha-lipoic acid

The present inventors conducted the following experiment to confirm whether the composition containing ferrous sulfate and alpha-lipoic acid inhibited stem cell aging.

The present inventors seeded 20,000/cm 2 Holoclone in 60 ødish and cultured in DMEM +10% FBS, 1% P/S for one day.

After washing the cultured cells twice with PBS, 0.02ml/cm2 of DMEM Phenol red-free media was filled and exposed to UVA 6J using a UV irradiator (BLX-254, VILBER Lourmat, France), and then Western blot (Western blot) confirmed the expression of senescence markers (P21, p-P53) in the cells.

The present inventors confirmed that the expression levels of the P21 and p-P53 were increased by Western blot results after exposing the Holoclone to UVA 6J.

The present inventors confirmed that after culturing the cells exposed to UVA 6J for 72 hours in Experimental Example (8), the amount of expression of P21 and p-P53 decreased as much as before exposure to UVA 6J (Fig. 13). UVA 6J, Experimental Example (8) conditions)

[Example 6] Confirmation of whitening effect by a composition containing ferrous sulfate and alpha-lipoic acid

The present inventors conducted the following experiment to confirm the whitening effect by the composition containing ferrous sulfate and α-Lipoic acid.

The present inventors treated with α-MSH to induce melanin pigment synthesis in mouse melanocytes (B16-F1) sold from Kyunghee University, and compared with the case where α-MSH was not treated, the α-MSH When treated with it was confirmed that the melanin level increased.

In the following [Table 5], when α-MSH was treated, the melanin level (negative control 2 in [Table 5]) was used as a standard (1.00 value), and when α-MSH was not treated ([Table 5] negative control 1), the relative melanin levels in the case of treating the experimental example (8) and the comparative examples were described.

When the α-MSH-treated B16-F1 was treated with Experimental Example (8), it was confirmed that the melanin level was relatively lower than that of Comparative Example 1 (α-Lipoic acid 200 μM). .

In addition, it was confirmed that the melanin level when the experimental example (8) was treated with α-MSH-treated B16-F1 was lower than that of the negative control group 2.

Through this, the present inventors confirmed that the whitening effect is better than the case of treatment with α-Lipoic acid alone when ferrous sulfate and α-Lipoic acid are combined.

α-MSH(-) α-MSH(+)
voice
Control 1 voice
Control 2 Experimental Example (8) Comparative Example 1 Comparative Example 2 Comparative Example 3 Ferrous sulfate (0.5 μM)+α-Lipoic acid (200 μM) α-Lipoic acid (200μM) RSV 50μM Arbutin 500μM 0.86 1.00
(standard) 0.79 0.93 0.79 0.91

[Example 7] Preparation of a cosmetic composition containing ferrous sulfate and α-Lipoic acid

The present inventors prepared a cosmetic composition of the composition shown in the following [Table 6].

The method for preparing the cosmetic composition is as follows.

1) Heat 2 to 7 (hereinafter, A) and 8 to 10 (hereinafter, B) in the following table to 75 °C, respectively.

2) Add A little by little to B and emulsify and stir at 7000 rpm.

3) After adding 1 and 11 in the following table, emulsify and stir for 2 minutes.

4) Degas the composition by stirring at 2500 rpm for 1 minute.

5) Soak the beaker containing the composition in ice water and cool it to 28-30 °C.

6) The prepared composition is allowed to stand at room temperature for 24 hours to stabilize.

turn ingredient weight% One Experimental Example (1) or Experimental Example (9) Up to 81 2 Stearic acid 1.0 3 Cetostearyl alcohol 0.7 4 Glycerin monostearate 0.5 5 Floating paraffin 5.0 6 Sorbitan monostearate 0.3 7 Squalane 3.5 8 Purified water To 100 9 Concentrated glycerin 6.5 10 Hyaluronic Acid Extract 0.5 11 Sepigel 350 0.7 12 incense Appropriate amount

[Example 8] Effect of Proliferation of Human Fibroblasts by Compositions Containing Various Components That Can be Used in Cell Culture

The present inventors have determined whether the cell proliferation effect can be exhibited by other components that can be used for cell culture in general (hereinafter, "cell culture component") other than ferrous sulfate and alpha-lipoic acid. In order to check whether or not, the following experiment was performed.

The present inventors conducted an experiment to confirm the cell proliferation effect using a composition in which a cell culture component other than ferrous sulfate and alpha-lipoic acid was mixed with a DMEM base medium. The reference content of each cell culture component in the composition is disclosed in each graph of FIGS. 14 to 17.

The cell culture components are L-Aspartic acid, L-Alanine, Proline, Pantothenic acid Ca-salt, Pyridoxine Hydrochloride, L-Ascorbic acid, L-asparagine monohydrate, L-Glutamic acid, D-biotin, zinc sulphate heptahydrate (Zinc Sulfate Heptahydrate), Riboflavin, and these were purchased from Sigma Aldrich. The DMEM basic medium is Hyclone?? Purchased from.

Each of the compositions was treated as a single product in human primary fibroblasts 4000/well for 48 hours. In addition, the present inventors treated the DMEM basal medium as a negative control, which was expressed as 0 condition in FIGS. 14 to 17.

When compared with the negative control, the proliferation effect of human primary fibroblasts by the compositions containing the cell culture component was not high (see FIGS. 14 to 17).

These examples are for illustrative purposes only, and those of ordinary skill in the art are not construed that the scope of the content disclosed by the present specification is limited by these examples. It will be obvious to you.

Claims (31)

Ferrous sulfate or a salt thereof; And
Lipoic acid or its salt
In the cosmetic composition comprising a medium containing as an active ingredient,
The ferrous sulfate or salt thereof is included in a concentration of 0.5 μM to 400 μM, based on the concentration in the medium,
The cosmetic composition, characterized in that the lipoic acid (Lipoic acid) or a salt thereof is contained in a concentration of 50 μM to 500 μM based on the concentration in the medium.
The method of claim 1,
The cosmetic composition, characterized in that the iron sulfate (Ferrous sulfate) is ferrous sulfate (iron (II) sulfate; FeSO ₄₎ .
The method of claim 2,
The ferrous sulfate (iron (II) sulfate; FeSO ₄ ) is an anhydrous salt (FeSO ₄ ), a tetrahydrate (FeSO _{4 ·} H ₂ O),
A cosmetic composition in the form of any one or more of five hydrated salts (FeSO ₄ H ₂ O), hexa hydrated salts (FeSO ₄ H ₂ O) and seven hydrates (FeSO ₄ H ₂ O).
The method of claim 1,
The lipoic acid is a cosmetic composition, characterized in that alpha-lipoic acid (α-Lipoic acid) or beta-lipoic acid (β-Lipoic acid).
The method of claim 4,
The alpha-lipoic acid is (R)-(+)- alpha-lipoic acid ((R)-(+)-α-lipoic
acid), (S)-(-)-alpha-lipoic acid ((S)-(-)-α-lipoic acid), or a mixture thereof.
The method of claim 1,
The ferrous sulfate or salt thereof is included in a concentration of 0.5 μM to 200 μM, based on the concentration in the medium,
The cosmetic composition, characterized in that the lipoic acid (Lipoic acid) or its salt is contained in a concentration of 100 μM to 500 μM based on the concentration in the medium.
The method of claim 1,
The ferrous sulfate or salt thereof is included in a concentration of 0.5 μM to 200 μM, based on the concentration in the medium,
The cosmetic composition, characterized in that the lipoic acid (Lipoic acid) or a salt thereof is contained in a concentration of 200 μM to 500 μM based on the concentration in the medium.
The method of claim 1,
The ferrous sulfate or a salt thereof is included in a concentration of 0.5 μM to 60 μM, based on the concentration in the medium,
The lipoic acid (Lipoic acid) or its salt is a cosmetic composition, characterized in that contained in a concentration of 200 μM to 400 μM based on the concentration in the medium.
The method of claim 1,
The medium is
Toxicity acting on cells including amino acids, sugars, vitamins, inorganic salts, lipids, cytokines, growth factors, extracellular matrix proteins, media, antioxidants, melanin pigment synthesis inhibitors, and melanin pigments A cosmetic composition, characterized in that it further comprises at least one of a substance and an exfoliating substance.
The method of claim 1,
The medium is
Glycine, Arginine, L-Glutamine, L-Glutamic Acid, Asparagine-H ₂ O, L-Alanine, Aspartic Acid (L-Aspartic acid), Cysteine hydrochloride (H ₂ O), histidine (L-Histidine), isoleucine (LIsoleucine), leucine (L-Leucine), lysine (L-Lysine), methionine ( L-Methionine), phenylalanine (L-Phenylalanine), serine (L-Serine), threonine (L-Threonine), tryptophan (L-Tryptophan), proline (L-Proline), valine (L-Valine), calcium plate Totenate (D-Calcium pantothenate), Folic Acid, Niacinamide, Pyridoxine hydrochloride, Riboflavin, Thiamine hydrochloride, Cyanocobalamine (Vitamin B12) ), Ascorbic Acid, Biotin, Inositol, Calcium Chloride (CaCl ₂ ), Magnesium Sulfate (MgSO ₄ )), Potassium Chloride ( KCl)), Sodium Bicarbonate (NaHCO ₃ )), Sodium Chloride (NaCl)), Sodium Phosphate monobasic (NaH ₂ PO ₄ -H ₂ O)), glucose (D-Glucose ( Dextrose)), sodium pyruvate (Sodium Pyruvate), adenosine (Adenosine), and a cosmetic composition comprising at least one of guanosine (Guanosine).
The method of claim 1,
The composition is
Flexible lotion, nourishing lotion, lotion, skin, body lotion, nutrition cream, massage cream, moisture cream, hand cream, eye cream, essence, cleansing foam, cleansing cream, cleansing water, pack, gel, patch, lipstick, makeup base, Powder, foundation, body cleanser, toothpaste, mouthwash, shampoo, conditioner, hair tonic, gel, mousse, hair-dressing agents such as hair essence, hairdressing agent, hair-dyed cosmetic composition, characterized in that it has a formulation selected from the group consisting of.
The method of claim 1,
The composition is a cosmetic composition exhibiting one or more of the following effects:
i) anti-aging effect;
ii) regenerative effect;
iii) wrinkle improvement effect;
iv) whitening effect; And
v) The effect of increasing skin elasticity.
The method of claim 1,
The composition is
Animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, lactose, aluminum hydroxide, calcium silicate, polyamide powder, water, ethanol, Isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyrene glycol oil, glycerol aliphatic ester, fatty acid ester of sorbitan, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, Aluminum metahydroxide, agar, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine , Fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, lanolin derivative, ethoxylated glycerol, fatty acid ester, stearic acid, cetostearyl alcohol, monostearic acid glycerin, squalane, concentrated glycerin, and hyaluronic acid extract. Cosmetic composition.
Ferrous sulfate or a salt thereof; And
In a cosmetic composition comprising as an active ingredient a medium containing lipoic acid or a salt thereof,
The cosmetic composition, characterized in that the ferrous sulfate or a salt thereof and the lipoic acid or a salt thereof are contained in a molar ratio of 1:1 to 1:10.
The method of claim 14,
The cosmetic composition, characterized in that the ferrous sulfate or a salt thereof and the lipoic acid or a salt thereof are contained in a molar ratio of 1:4 to 1:8.
Ferrous sulfate or a salt thereof at a concentration of 0.5 μM to 400 μM; And 50 μM to 500 μM Lipoic acid (Lipoic acid) or a medium for cell culture comprising a salt thereof.
Cell culture medium, characterized in that containing iron sulfate (Ferrous sulfate) and the lipoic acid (Lipoic acid) in a molar ratio of 1:1 to 1:10.
The method of claim 16 or 17,
The cells include skin epidermal cells and skin dermal cells; Their progenitor cells; And a medium for cell culture, characterized in that any one of these stem cells.
The method of claim 16 or 17,
The medium is
Toxicity acting on cells including amino acids, sugars, vitamins, inorganic salts, lipids, cytokines, growth factors, extracellular matrix proteins, media, antioxidants, substances that inhibit melanin synthesis, melanin pigments A medium for cell culture, characterized in that it further comprises one or more of a substance and a keratin exfoliation substance.
The method of claim 16 or 17,
The medium is Glycine, Arginine, L-Glutamine, L-Glutamic Acid, Asparagine-H2O, Alanine, and Aspa. L-Aspartic acid, L-Cysteine hydrochloride-H2O, L-Histidine, LIsoleucine, L-Leucine, L-Lysine, and methionine -Methionine), phenylalanine (L Phenylalanine), serine (LSerine), threonine (L-Threonine), tryptophan (L-Tryptophan), proline (L-Proline), valine LL-Valine), calcium pantothenate (D- Calcium pantothenate), FolicAcid, Niacinamide, Pyridoxinehydrochloride, Riboflavin, Thiaminehydrochloride, Cyanocobalamine (Vitamin B12), Ascorbic Acid , Biotin, Inositol, Calcium Chloride (CaCl2), Magnesium Sulfate (MgSO4)), Potassium Chloride (KCl)), Sodium Bicarbonate ( NaHCO3)), Sodium Chloride (NaCl), Sodium Phosphatemonobasic (NaH2PO4-H2O)), Glucose (D-Glucose (Dextrose)), Sodium Pyruvate, Adenosine and Guanosine (Guanosine), characterized in that the medium for cell culture comprising one or more of. delete delete delete delete delete delete delete delete delete delete delete

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