KR20200034331A - A composition for activating a skin cell and uses thereof - Google Patents

Abstract

The present invention relates to a composition for activating skin stem cells. The composition can include one or more components of ferrous sulfate and lipoic acid. Skin cell activation effects can be derived by the composition provided in the present invention, and the skin cells can be skin stem cells. Stem cell proliferation and/or division are actively caused by the skin stem cell activation, and ultimately, aged skin can be regenerated. In addition, the skin whitening effect can be shown by the composition.

Description

A composition for activating a skin cell and uses thereof

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

Recently, interest in stem cell cosmetics is increasing, and general stem cell cosmetics refer to cosmetics that use stem cell culture fluid as a component, which contains components secreted during stem cell culture.

Cosmetics containing components capable of increasing the activity of the stem cell itself as well as such stem cell culture fluids are being developed, and in this case, there is an advantage of manufacturing cosmetics through low-cost raw materials and simple processes.

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

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

According to an aspect of the technology disclosed by the present application, in order to achieve the above-described problem, ferrous sulfate; And lipoic acid (Lipoic acid) or a salt thereof; provides a composition for skin cell activation comprising a.

The ferrous sulfate may be ferrous sulfate (iron sulfate (II); FeSO ₄ ), for example, tetrahydrate (FeSO ₄ · 4H ₂ O), pentahydrate (FeSO ₄ · 5H ₂ O ), Hexahydrate (FeSO ₄ · 6H ₂ O) and may be one or more of 7 hydrate (FeSO ₄ · 7H ₂ O).

Further, the lipoic acid may be alpha-lipoic acid or beta-lipoic acid, and the alpha-lipoic acid may be (R)-(+)-alphalipoic acid. ((R)-(+)-α-lipoic acid), (S)-(-)-alphafoic acid ((S)-(-)-α-lipoic acid) or mixtures thereof.

The skin may be dermal or epiderma, and the 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 their salts, which act as main components, amino acids, sugars, vitamins, inorganic salts, lipids, cytokines, and growth factors ( Growth factor), extracellular matrix protein, media, antioxidants, melanin pigment synthesis inhibitors, toxic substances acting on cells containing melanin pigments and one or more of the exfoliation material may be further included.

The above-mentioned composition for activating skin cells may function as i) cell proliferation effect and / or cell aging regulator expression suppression function. The aging regulator is a CDK inhibitor (Cyclic-dependent Kinase) (CDK) inhibitor, and 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; Provided is a cosmetic composition for skin cell activation comprising at least one of the above.

The cosmetic composition is glycine (Glycine), arginine (L-Arginine), glutamine (L-Glutamine), glutamic acid (L-Glutamic Acid), asparagine (L-Asparagine-H2O), alanine (L-Alanine ), Aspartic acid, L-Cysteine hydrochloride-H2O, L-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) , Calcium pantothenate, Folic Acid, Niacinamide, Pyridoxine hydrochloride, Riboflavin, Thiamine hydrochloride, Cyanocobalamine (Vitamin B12), Ascorbic Acid, Biotin, Inositol, Calcium Chloride (CaCl2), Mag Nesium Sulfate (MgSO4), Potassium Chloride (KCl), Sodium Bicarbonate (NaHCO3), Sodium Chloride (NaCl), Sodium Phosphate monobasic (NaH2PO4- H2O)), glucose (D-Glucose (Dextrose)), sodium pyruvate (Sodium Pyruvate), adenosine (Adenosine) and guanosine (Guanosine).

The cosmetics include softening 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 essence, and other hairdressing agents, may be a formulation selected from the group consisting of hair and hair dyes.

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

The cell activation effect may be exhibited by the composition disclosed by the present specification, and ultimately, the skin condition improving effect may also be exhibited.

The cell activation effect may be an effect of inhibiting cell proliferation, differentiation, or cell aging. In addition, as a skin condition improving effect, wrinkle improvement effect, anti-aging effect, skin elasticity effect, and whitening effect may be exhibited.

1 is a human fibroblast (Human Primary) by Mix I and Mix III containing Ferrous sulfate and Alpha-Lipoic acid in the modified medium and other components included in the modified medium according to one embodiment. Fibroblast).
Figure 2 (a) shows the proliferation of human fibroblasts (Human Primary Fibroblast) by a composition containing alpha-lipoic acid (α-Lipoic acid).
Figure 2 (b) shows the proliferation of human fibroblasts (Human Primary Fibroblast) by a composition containing iron sulfate (Ferrous sulfate).
Figure 3 shows the proliferation effect of human fibroblasts (Human Primary Fibroblast) by the composition (Experimental Example (a) to Experimental Example (i)) containing ferrous sulfate and alpha-lipoic acid.
Figure 4 shows the results of confirming the dermal stem cell markers (CD29 (+), CD34 (-)) of the Holoclone and the proliferation effect of the Holoclone.
Figure 5 shows the results of confirming the dermal stem cell markers (CD44 (+), CD90 (+), CD105 (+)) of the holoclone.
FIG. 6 shows the degree of growth of holoclone by a composition containing ferrous sulfate.
7 shows the degree of proliferation of Holoclone by the composition containing alpha-Lipoic acid.
FIG. 8 shows the effect of proliferation of Holoclone by a composition (Experimental Example (1) to Experimental Example (9)) containing ferrous sulfate and alpha-lipoic acid.
9 is a graph comparing the proliferative effect of human primary fibroblasts and Holoclone according to Experimental Example (8).
10 shows the proliferation degree of bone marrow-derived mesenchymal stem cells according to Experimental Examples (1) to (9).
11 shows the degree of proliferation of adipose-derived mesenchymal stem cells according to Experimental Examples (1) to (9).
12 shows the effect of wound healing (Wound Healing) by the composition 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).
FIG. 14 shows the degree of human primary fibroblast proliferation by a composition comprising L-Aspartic acid, L-Alanine or Proline in DMEM basic medium.
FIG. 15 shows human primary fibroblast proliferation by a composition comprising calcium pantothenic acid (Pantothenic acid Ca-salt), pyridoxine hydrochloride or L-Ascorbic acid in DMEM basic medium. Degree.
FIG. 16 is a human fibroblast (Human) by a composition comprising L-asparagine monohydrate, L-Glutamic acid or D-biotin in DMEM basic medium. Primary Fibroblast).
FIG. 17 shows the degree of human primary fibroblast proliferation by a composition comprising 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 tissue that protects the body's organs and muscles from external stimuli, which can be damaged by various external factors, and the skin consists of the epidermis, dermis and subcutaneous layers. ) Is composed of a stratum corneum layer, a granular layer, a polar layer, and a base layer.

The epidermis includes keratinocytes, melanocytes, Langerhans cells, and Merkels cells, and keratinocytes are the majority of epidermal cells (about 95%). Occupies.

The dermis is a layer of skin that exists between the epidermis and the subcutaneous 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 is composed of adipose tissue and connective tissue. The subcutaneous fat layer contains adipocytes.

The subcutaneous fat layer (Hypodermis) is the thickest layer of the skin, and is composed of adipose tissue and connective tissue. The subcutaneous fat layer contains adipocytes.

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

The term "primary cell" refers to cells cultured separately from living tissue.

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

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

The term "adult stem cell" is an undifferentiated cell that exists between differentiated cells of a specific tissue or organ, and can serve to replace and repair damaged tissue. Adult stem cells can differentiate only to each target organ.

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

The term “Holoclone” is well known to those skilled in the art as stem cells forming colonies. (Nature Medicine volume 12, pages 1397-1402 (2006))

In this specification, the term “holoclone” and the term “stem cell” may be used interchangeably.

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

The term "medium" is a medium containing nutrients for culturing cells (Culture), 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" means to proliferate or maintain the cells, and the culture may include both adherent culture and suspension culture.

The term "Proliferation" may mean that the number of cells increases due to the division of cells, and the proliferation of the cells includes both monolayer growth or multi-layer growth.

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

For example, the basic medium may be 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 components of the basic medium are modified.

For example, the modified medium may be a case in which some of the components of the basic medium are removed. For another example, the modified medium may be a case in which a component other than the component of the basic medium is added to a component of the basic medium. For another example, the modified medium may be a case in which some of the components of the basic medium are removed and components other than the components of the basic medium are added.

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

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

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

Main components of cell activation composition

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

The composition may include one or more of ferrous sulfate and lipoic acid as essential components.

The ferrous sulfate includes green vitriol, iron vitriol, coppereras, melanterite, zomolnokite, iron sulfate, iron sulphate, and ferrous sulfate. (Ferrosulfate), FeSO4, Green salts (Green salts).

The ferrous sulfate is ferrous sulfate (iron (II) sulfate; FeSO ₄ ), ferrous sulfate (iron (II) 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 beard (FeSO ₄ · 7H ₂ O).

In another embodiment, in the case of ferric sulfate, anhydrous salt (Fe ₂ (SO ₄ ) ₃ ), trihydrate (Fe ₂ (SO ₄ ) ₃ · 3H ₂ O), hexahydrate (Fe ₂ (SO ₄ ) ₃ 6H ₂ O), 7 bead (Fe ₂ (SO ₄ ) ₃ · 7H ₂ O), 7.5 beard (Fe ₂ (SO ₄ ) ₃ · 7.5H ₂ O), 9 beard (Fe ₂ (SO ₄ ) ₃ · It may be 9H ₂ O), 10 hydrate (Fe ₂ (SO ₄ ) ₃ · 10H ₂ O) or 12 hydrate (Fe ₂ (SO ₄ ) ₃ · 12H ₂ O).

As a typical example, ferrous sulfate may be 7 beta (FeSO ₄ · 7H ₂ O).

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

A person skilled in the art can arbitrarily adjust the content of the ferrous sulfate to include in the composition.

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

In one embodiment, ferrous sulfate may be included from about 0.5 μM to 100 μM.

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

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

For example, ferrous sulfate may be included from about 100 μM to 500 μM. For example, ferrous sulfate may be included from about 500 μM to 1 mM.

Ferrous sulfate, which may be included as a main component in the present specification, is generally known as a material that functions to supply oxygen or supplement iron.

Lipoic acid (Lipoic acid) is a fatty acid having 8 carbons and 2 sulfurs, and is produced in a small amount in the human body.

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

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

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

In another embodiment, alpha-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-dithiore-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 mixtures thereof. For example, β-Lipoic acid ((R)-(+)-betalipoic acid ((R)-(+)-β-Lipoic acid), (S)-(-)-betalipoic acid (( S)-(-)-β-Lipoic acid) or mixtures thereof.

A person skilled in the art can arbitrarily adjust the concentration of the lipoic acid to include it in the composition.

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

In one embodiment, lipoic acid (Lipoic acid) may be included from about 0.5μM to 100μM.

For example, lipoic acid may be included in the range of about 0.5 μM to 50 μM. For example, lipoic acid may be included from about 50 μM to 100 μM. In a specific embodiment, lipoic acid (Lipoic acid) may be included about 0.5μM.

In another embodiment, lipoic acid may be included from about 100 μM to 300 μM.

For example, lipoic acid may be included from about 100 μM to 250 μM. In a specific embodiment, lipoic acid (Lipoic acid) may be included about 200μM. For example, lipoic acid may be included from about 250 μM to 300 μM.

In another embodiment, the lipoic acid (Lipoic acid) may be included from about 300μM to 500μM.

For example, lipoic acid may be included in the range of about 300 μM to 450 μM. Specifically, lipoic acid may include about 400 μM. For example, lipoic acid may be included from about 450 μM to 500 μM.

In another embodiment, the lipoic acid (Lipoic acid) may be included from about 500μM to 1mM.

Lipoic acid, which can be included as a main 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 fats, activate mitochondria, accelerate 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 main ingredients of composition

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

Specifically, the lipoic acid (Lipoic acid) may play a major role in increasing the activity of the cell. The combination of the ferrous sulfate and the lipoic acid may increase the activity of cells.

The term "cell activity" is

i) cell proliferation,

ii) cell regeneration,

iii) preventing the aging of existing cells or

iv) cell whitening effect.

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

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

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

The cells targeted for the increase in activity may be skin cells, and the skin cells may include cells present in the epidermis and 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 fully differentiated cells, progenitor cells, and stem cells.

The term "progenitor cell" is an intermediate stage cell between a stem cell and a fully differentiated cell, and can be differentiated into a specific cell and can divide only a limited number of times unlike a stem cell.

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

For example, in the case of skin aging or damage, not only fully differentiated cells among the skin cells, but also stem cells have reduced activity, and stem cells with reduced activity are capable of producing autologous cells compared to stem cells without reduced activity. This drops to about a third. As the number of autogenous cells decreases, the regenerative capacity of the skin decreases, and a typical example is wrinkles.

Therefore, it is necessary to increase cell activity for skin regeneration, anti-aging or maintaining skin elasticity.

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

For example, cells targeted for increased activity may be muscle cells, nerve cells, germ cells, immune cells or bone cells.

In addition, the degree of differentiation of cells that are the target of the increase in activity is not limited.

For example, fully differentiated cells, progenitor cells, or stem cells can all be targeted for 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 components other than ferrous sulfate and lipoic acid.

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

In addition, the optional components may be formulated within a range not impairing the object and effect of the present invention.

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

For example, amino acids are glycine, alginine, alginine hydrochloride glutamine, histidine, isoleucine, serine, leucine , Lysine, Methionine, Phenylalanine, Threonine, Tryptophan, Valine, Tyrosine, Cysteine, L-Cysteine hydrochloride H2O), Proline, Asparagine-H2O, Asparagine, Glutamic acid, Aspartic Acid, Histidine hydrochloride-H2O, And alanine (Alanine) may be selected from one or more groups.

 For example, one or more sugars may be selected from the group consisting of glucose, galactose, ribose, fructose, sucrose, lactose, and maltose. You can.

For example, vitamins are a group 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 a combination thereof. One or more may be selected from the group consisting of precursors.

Specifically, vitamins include retinol, retinal, retinoid, carotenoid, thiamin, thiamine hydrochloride, riboflavin, niacin, niacinamide, pantothenic acid, pyridoxine, pyridoxine H. pyridoxine hydrochloride Doxamine, Pyridoxal, Biotin, Folic Acid, Folic Acid, Cyanocobalamin (Vitamin B12), Hydroxycobalamin, Methylcobalamin, Ascorbic Acid, Ergocalciferol, Cholecalciferol, Tocopherol , Tocotrienol, phylloquinone, menaquinone, D-Calcium pantothenate, polyacid, riboflavin and i-Inositol. .

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 to.

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

In other embodiments, the protein may be a protein or extracellular matrix protein involved in the cellular signaling system, and these may be substances secreted from the cell.

For example, 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 (Cytokine) or a growth factor (Growth factor), but is not limited thereto.

Specifically, 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 and TGFβ2.

Specifically, growth factors include insulin-like growth factor (IGF), epidermal growth factor (EGF), fibroblast growth fac-tor (FGF), and nerve growth. Nerve growth factor (NGF), trans-forming growth factor (TGF), platelet-derived growth fac-tor (PDGF), bone-derived growth factor (BGF), BDF) and a colony stimulation factor (CSF) may be selected from one or more, but is not limited thereto.

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

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

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

The composition comprising 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 one or more of 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 a component of a cosmetic carrier may be composed of components permitted as cosmetic raw materials notified by the Ministry of Food and Drug Safety.

Specifically, when the formulation of the cosmetic is a paste, gel, or cream, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, and zinc oxide as cosmetic carrier ingredients One or more groups may be selected from, but are not limited to.

Specifically, when the formulation of the cosmetic is a powder or a 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, one or more propellants may be selected from the group consisting of chlorofluorohydrocarbon, propane, butane and dimethyl ether.

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

Specifically, when the formulation of the cosmetic is a suspension, liquid diluents such as water, ethanol or propylene glycol as a cosmetic carrier component, suspensions such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, One or more selected from the group consisting of microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tracant.

Specifically, when the cosmetic formulation is a surfactant-containing cleansing agent, as a cosmetic carrier component, an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyl taurate, sarcosinate, One or more may be selected from the group consisting of fatty acid amide ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives and ethoxylated glycerol fatty acid esters.

For example, a composition that can be used as a component of a pharmaceutical carrier may be composed of components permitted as pharmaceutical raw materials notified by the Ministry of Food and Drug Safety.

Specifically, pharmaceutical carriers included in pharmaceuticals include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose , Water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like.

In addition, lubricants, wetting agents, sweetening agents, flavoring agents, emulsifying agents, suspending agents, preservatives, etc. may be further included in addition to the above components.

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

More specifically, for suppositories for parenteral administration, one or more are selected from the group consisting of Witepsol, Macrogol, Tween 61, cacao butter, laurin and glycerogelatin as pharmaceutical carrier components. It may be, but is not limited to this.

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 excipients, 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 preparation for oral administration, water, a liquid paraffin, or an excipient may be included as a pharmaceutical carrier component, but is not limited thereto. More specifically, excipients can be wetting agents, sweetening agents, fragrances or preservatives.

In another embodiment, the ingredients that can be used for skin improvement may be antioxidants, melanin pigment synthesis inhibitors, toxic substances acting on cells containing melanin pigments, or substances that exfoliate keratin, but are 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, isoflavones may be genistein and daidzein. More specifically, the vitamin may be tocopherol.

Specifically, the melanin pigment synthesis inhibitor may be a tyrosinase inhibitor or a substance that removes free radicals. More specifically, the tyrosinase inhibitor may be mulberry extract, arbutin, licorice extract, vitamin C, kojic acid, or alpha bisabolol (α-Bisabolol), but is not limited thereto.

More specifically, the substance that removes free radicals may be an antioxidant.

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

Specifically, the material that exfoliates keratin may be alpha hydroxy acid (AHA), but is not limited thereto.

In another embodiment, the composition comprising one or more of ferrous sulfate and lipoic acid provided herein may further include other cell energy sources, antibiotics, flavoring agents, preservatives or pigments. 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 aforementioned components.

Effect of the composition for cell activation

By the present specification, various compositions that cause cell activation effects can be provided.

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

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

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

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 stem cells present in the dermis of the skin may be increased by the composition provided herein.

The activity of differentiated cells differentiated by the composition provided herein may be increased.

The activity of skin stem cells or progenitor cells may be increased by the composition provided herein.

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, the activity of all differentiated cells, progenitor cells, or stem cells may be increased.

(1) Effect of cell activation composition-cell proliferation

The proliferation effect of cells can be increased by the composition.

According to some embodiments provided in the present application, when compared to the cell proliferation effect by a composition that does not contain ferrous sulfate and lipoic acid, the ferrous sulfate and / or lipoic acid ( Cell proliferation effect by the composition containing Lipoic acid) may be about 2 times higher.

For example, compared to the effect of proliferation of skin stem cells by a composition that does not contain ferrous sulfate and lipoic acid, the ferrous sulfate and lipoic acid are combined in a specific concentration. Skin cell proliferation effect by the composition comprising as may be high.

Specifically, the effect of skin cell proliferation by 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 ferrous sulfate and lipoic acid ( Lipoic acid) may be higher than the proliferation effect of the composition.

For another example, skin cell proliferation effect by a composition comprising ferrous sulfate and lipoic acid in a specific ratio may be high.

Specifically, the effect of skin cell proliferation by the composition containing ferrous sulfate and lipoic acid in a 1: 1 to 1:10 molar concentration ratio is iron sulfate (Ferrous sulfate) and lipoic acid (Lipoic acid). It may be higher than the skin cell proliferation effect by the composition not included.

Depending on the type of skin cells, the molar concentration 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 large number of fibroblasts, a high cell proliferation effect when ferrous sulfate and lipoic acid are contained in a molar ratio of 1: 1 to 1: 2 May appear.

As another example, in the case of skin cells containing a large number of skin stem cells, iron sulfate (Ferrous sulfate) and lipoic acid (Lipoic acid) are contained in a ratio of 1: 4 to 1: 8 molar concentration. Proliferative effects may be present.

According to some embodiments provided in the present application, the cell proliferation effect by the composition comprising a combination of ferrous sulfate and lipoic acid is only the ferrous sulfate without lipoic aicd. It may be about 2 times higher than the cell proliferation effect by the composition comprising.

In addition, the cell proliferation effect by the composition containing a combination of ferrous sulfate and lipoic acid is a 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 proliferative effect of skin stem cells by a composition containing ferrous sulfate and / or lipoic acid may be exhibited.

For example, compared to the proliferation effect of bone marrow-derived-mesenchymal stem cells or adipose-derived mesenchymal stem cells, proliferation of skin stem cells by a composition comprising ferrous sulfate and / or lipoic acid The effect may be 1.5 to 3 times higher. Specifically, compared to other adult stem cells other than skin stem cells, the proliferation effect of skin stem cells by a 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 in a composition comprising the ferrous sulfate or the lipoic acid, respectively. It may be higher than the proliferation effect.

For example, compared to the effect of proliferation of skin stem cells by a composition that does not contain ferrous sulfate and lipoic acid, the ferrous sulfate and lipoic acid are combined in a specific concentration. Skin stem cell proliferation effect by the composition comprising as may be high.

Specifically, the effect of skin stem cell proliferation by the 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 is ferrous sulfate and lipoic acid. acid) may be higher than the proliferation effect of the composition.

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 is ferrous sulfate and lipoic acid acid) may be about 2 to 4 times higher than the proliferation effect of the composition.

More specifically, the effect of skin stem cell proliferation by the composition containing about 0.5 μM to about 10 μM of ferrous sulfate and about 100 μM to about 500 μM of lipoic acid is ferrous sulfate and lipoic acid ( Lipoic acid) may be about 2 to 3 times higher than the proliferation effect by the composition containing no.

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 ferrous sulfate and lipoic acid. ) May be about 2 to 3 times higher than the proliferation effect by the composition not containing.

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

In 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 ferrous sulfate and lipoic acid. ) May be about 2 to 3 times higher than the proliferation effect by the composition not containing.

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

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 is ferrous sulfate and lipoic acid acid) may be about 3 to 4 times higher than the proliferation effect of the composition.

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 ferrous sulfate and lipoic acid. It may be about 3 to 4 times higher than the proliferation effect by the composition containing no.

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

In 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 ferrous sulfate and lipoic acid. It may be about 3 to 4 times higher than the proliferation effect by the composition containing no.

In another example, the effect of proliferation of skin stem cells by a composition containing about 50 μM of ferrous sulfate and about 400 μM of lipoic acid is due to a composition that does not contain 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 the composition containing about 60 μM to about 200 μM of ferrous sulfate and about 100 μM to about 500 μM of lipoic acid is ferrous sulfate and lipoic acid. acid) may be about 3 to 4 times higher than the proliferation effect of the composition.

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 ferrous sulfate and lipoic acid. It may be about 3 to 4 times higher than the proliferation effect by the composition containing no.

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

As another example, the effect of proliferation of skin stem cells 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 ferrous sulfate and lipoic acid. It may be about 3 to 4 times higher than the proliferation effect by the composition containing no.

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

For another example, skin stem cell proliferation effect by a composition containing ferrous sulfate and lipoic acid in a specific ratio may be high.

Specifically, the effect of proliferation of skin stem cells by the composition containing ferrous sulfate and lipoic acid in a 1: 1 to 1:10 molar concentration ratio is ferrous sulfate and lipoic acid. It may be higher than the skin stem cell proliferation effect by the composition containing no.

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

More specifically, the effect of proliferation of 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 can be about 3 times to about 4 times higher than the skin stem cell (Skin stem cell) proliferation effect by the composition containing no lipoic acid (Lipoic acid).

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

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

For 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 the ferrous sulfate without lipoic aicd. It can be about 4 times higher than the skin stem cell proliferation effect by.

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

For another example, skin stem cell proliferation effect by a composition comprising a combination of ferrous sulfate and lipoic acid is about 1.5 to about 2 times higher than the proliferation effect of primary fibroblast You can.

According to some embodiments provided in the present application, even when the composition provided by some embodiments of the present application does not include animal-derived serum, a cell proliferation effect as high as when treated with animal-derived serum may be exhibited.

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

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

Specifically, when the composition is treated on 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 serum of the soybean can be FBS (Fetal Bovine Serum), BCS (Bovine Calf Serum) or FCS (Fetal Calf Serum). More specifically, the animal-derived serum may be 10% FBS.

(2) Effect of cell activation composition-cell regeneration

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

In one embodiment, the cell regeneration effect by the composition containing the ferrous sulfate and lipoic acid is a cell by the composition that does not contain the ferrous sulfate and the lipoic acid It can 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 1: 2 to 1: 8 molar concentration ratio 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 a composition comprising ferrous sulfate and lipoic acid is a regenerating effect by a composition comprising the ferrous sulfate or the lipoic acid, respectively. It can be higher.

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

For another example, the cell regeneration effect by a composition comprising a combination of ferrous sulfate and lipoic acid is caused by a composition containing only the lipoic acid without the ferrous sulfate. It may be about 2 times to about 4 times higher than the cell regeneration effect.

(3) Effect of cell activation composition-Prevent cell aging

Cells may be prevented from aging by the composition, and may exhibit cell anti-aging effects through suppression of aging regulator expression, but are not limited thereto.

The term "aging regulator" is a substance that inhibits the action of an enzyme that regulates the cell cycle. As the expression of the aging regulator is suppressed, aging may be prevented.

In one embodiment, expression of the aging regulator may be inhibited by the composition provided herein.

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

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

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

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

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

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

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

The whitening effect may be an effect of inhibiting the synthesis of melanin pigment in cells, decomposition of melanin produced in cells, destruction of cells containing melanin pigment, or exfoliation.

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

For example, the composition provided herein may be a material that serves to scavenge free radicals or free radicals. For example, the composition provided herein may be a substance that serves to destroy cells containing melanin pigment. For example, the composition provided herein may be a material that serves to exfoliate keratin.

Specifically, the composition provided herein may exhibit a higher whitening effect than arbutin, niacinamide, kojic acid, and / or RSV, which are well-known as substances showing a whitening effect. have.

Specifically, the cell whitening effect by the composition containing the ferrous sulfate and lipoic acid provided in the present specification is in the composition containing only the lipoic acid without the ferrous sulfate It may be higher than the cell whitening effect.

Specifically, the cell whitening effect by the composition comprising the ferrous sulfate and lipoic acid provided in the present specification is in a composition containing only the ferrous sulfate without the lipoic acid It may be higher than the cell whitening effect.

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

The skin condition improving effect may be exhibited by the composition provided herein, and the condition of the epidermis, dermis or subcutaneous layer may be improved.

The skin condition improving effect may be exhibited through cell activation as well as inhibition of specific substances, but is not limited thereto.

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

For example, the proliferation effect of epidermal cells by the composition provided herein may be exhibited, 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 the extracellular matrix component of the dermis or the effect of proliferating the dermal cells, thereby improving the 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, the skin whitening effect may be exhibited by the composition provided herein.

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

For another example, the melanin pigment produced in the cell 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, nourishing lotion, lotion, skin, body lotion, nourishing cream, massage cream, moisture cream, sun cream, hand cream, eye cream, essence, cleansing foam, cleansing cream, cleansing water, pack, It can be formulated as a gel, oil or patch, but is not limited thereto.

For example, makeup cosmetics may be formulated as lipstick, makeup base, powder or foundation, but are not limited thereto.

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

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

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

In one embodiment, the medium for cell culture may include one or more of iron sulfate (Ferrous sulfate) and lipoic acid (Lipoic acid), in general, a component that can be added during cell culture may be additionally included.

The cosmetic composition may include a cell culture solution.

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

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

In one embodiment, the peptide may be a cytokine (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 can be a growth factor.

For example, growth factors include insulin-like growth factor (IGF), epidermal growth factor (EGF), fibroblast growth fac-tor (FGF), and nerves. May be growth factor (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.

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

Ultimately, an anti-aging effect, regenerating 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 obvious to those skilled in the art.

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

In one embodiment, emulsification is a high temperature emulsification method, inversion emulsification method, DSA emulsification method, PIT emulsification method, or liquid crystal emulsification. It may be by the method (LC Emulsification Method), but is not limited thereto.

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

In another embodiment, the dispersion (Dispersion) may be to mix and dissolve by adding a dispersion medium and rotating the stirrer.

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 raw materials notified by the Ministry of Food and Drug Safety.

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 preparation, an injection, an eye drop, an intravenous solution, an external solution, an ointment, a patch, a solid or a liquid, but is not limited thereto.

For example, tablets, tablets, tablets, capsules, powders, granules, pills, troches, syrups, inhalants, oral disintegrating tablets, chewable tablets, foaming tablets, dispersion tablets, dissolving tablets, foam granules, multi-drugs, oral cavity It may be a dissolving tablet, sublingual tablet, baccal tablet, adherent tablet, gum agent, oral dissolving film, inhalation powder, nasal powder, ex-agent or oral jelly.

For example, injections may be injections, powder injections, infusions, lyophilized injections, implants, sustained injections, peritoneal dialysis agents, perfusion or dialysis agents.

For example, oral solutions, oral solutions, syrups, emulsions, suspensions, elixirs, lemonades, tinctures, fluid extracts, alcohols, fragrances, premise, acupuncture, inhalants, inhalants, extracts, oral preparations It may be a spray agent for inhalation, an inhaled aerosol, a nasal drops, a gargle or an oral jelly.

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

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

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

For example, the solid agent may be an externally used acid, an inhalant, an inhaled powder, an externally used solid, a gargle, a hemodialysis agent, a dialysis agent, an ear drops, 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 differentiated cells, a medium for culturing progenitor cells, or a medium for culturing stem cells.

For example, fully differentiated cells may be muscle cells, nerve cells, skin cells, germ cells, immune cells, or bone cells, but are 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 (Keratinocyte) or melanocytes (Melanocyte).

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

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

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

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

As described above, the cell activation effect may be exhibited by a composition comprising at least one of ferrous sulfate and lipoic acid provided herein.

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 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 medium for cell culture, which includes at least one of the ferrous sulfate and lipoic acid as an essential component, may also exhibit a cell activation effect.

Hereinafter, the composition and the effects provided by 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 a composition provided by the present specification by including ferrous sulfate and lipoic acid in various contents in a modified medium in which some of the components of a commercial medium were changed.

The modified medium includes the components shown in [Table 1] below.

The present inventors custom-made and used the modified medium at Weljin Co., Ltd. In the embodiment disclosed herein, the content of the ferrous sulfate and lipoic acid in the medium is 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 Polyacid 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 HLC One 67-03-08 (Thiamine HCl) Vitamin B12 Cyanocobalamin 1.36 68-19-9 (Cyanocobalamin) Ascorbic Acid Ascorbic 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 alpha-lipoic acid

The present inventors prepared compositions Mix II and Ferric sulfate and alpha lipoic acid contained 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.

Table 2 below discloses the components excluding the modified medium components among the components of MixI, MixII, and MixIII.

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) 6Mo7O244H2O)  NH4 0.02uM 0.1uM

1-2: Preparation of a composition containing ferrous sulfate and alpha-lipoic acid (hereinafter, Experimental Example (a) to Experimental Example (i))

In addition, the present inventors made a composition (Experimental Example (a) to Experimental Example (i)) by varying the contents of ferrous sulfate and alpha-lipoic acid in the modified medium for detailed concentration evaluation. It was produced.

 Table 3 below discloses the molar concentrations of ferrous sulfate and alpha-lipoic acid included in each composition (Experimental Example (a) to Experimental Example (i)).

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 alpha-lipoic acid (Experimental Example (1) to Experimental Example (9))

The present inventors prepared nine compositions (Experimental Example (1) to Experimental Example (9)) for processing on the Holoclone having stem cell properties, and the nine compositions had a low concentration of Ferrous Sulfate. ).

In Table 4, the molar concentrations of ferrous sulfate and alpha-lipoic acid included in each composition (Experimental Example (1) to Experimental Example (9)) were 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] Fibroblast proliferation effect by treatment with a composition containing a combination of ferrous sulfate and alpha-lipoic acid

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

2-1: Fibroblast proliferation effect by MixII treatment

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

After washing the cultured human fibroblasts (Human Primary Fibroblast) twice with PBS, MixI, MixII and MixIII were equally dispensed into wells for 100ul, and cultured for 72 hours to confirm the proliferation effect. Substances constituting the Mix I, Mix II and Mix III were diluted in 1M or 0.5M stock concentration to a Low or High concentration, respectively, and treated to the human primary fibroblast.

The proliferation effect was measured using a CCK-8 assay, diluting the CCK-8 solution in DMEM medium without FBS by 1/10, dispensing it into each well for 100ul, reacting in an incubator at 37 ℃ for 1-4 hours, and then 450nm. Measured at absorbance.

 The present inventors have proliferation effect of about 2 times or more when in the MixII-High condition than in the condition that does not contain Ferrous sulfate and Alpha-Lipoic acid (0 condition in FIG. 1 or MixI, MixIII condition). It was confirmed that appears (see Figure 1).

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

The present inventors have the effect of cell proliferation by the composition containing ferrous sulfate in the modified medium and the composition containing alpha-Lipoic acid in the modified medium in the same manner as the MixI, MixII and MixIII treatment methods described above. Confirmed.

The present inventors, as shown in Figure 2 (a), when the cell proliferation effect by the composition containing the alpha-lipoic acid (α-Lipoic acid) in the modified medium modified medium + 10% FBS treatment (Fig. 2 ( PC condition of a); hereinafter, it was confirmed that it is not as high as in the 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 (Ferrous sulfate) in the modified medium is also a positive control (PC condition of Figure 2 (b)) It was not as high, 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: Human fibroblast proliferation effect according to Experimental Examples (a) to (i)

The present inventors treated Experimental Example (a) to Experimental Example (i) in the same manner as the MixI, MixII and MixIII treatment methods described above, respectively, to the human fibroblasts (Human Primary Fibroblast) to confirm the cell proliferation effect.

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

Furthermore, when the human fibroblasts (Human Primary Fibroblast) are treated with the Experimental Example (e), Experimental Example (f), Experimental Example (h) and Experimental Example (i), the cell proliferation is about 1.7 times higher than the negative control. The effect was confirmed.

The present inventors, rather than the cell proliferation effect by the composition containing the above-described alpha-lipoic acid (α-Lipoic acid) in the 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 by (see FIGS. 2 (a) and 3) is high.

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

The present inventors tested the composition containing the ferrous sulfate and alpha-lipoic acid ([Table 4]) to confirm that the proliferation effect is due to skin stem cell proliferation. Examples (1) to Experimental Example (9)) were treated with a Holoclone isolated from human primary fibroblasts.

[Example 3] Skin stem cell proliferation effect by treatment with a composition containing iron sulfate (Ferrous sulfate) and alpha-lipoic acid (α-Lipoic acid)

3-1: Skin stem cell obtained

(1) Holoclone separation from human dermal derived fibroblasts

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

Therefore, the present inventors conducted an experiment to investigate the proliferation effect of the Holocalone (Holoclone) in order to confirm the proliferation of skin stem cells in the composition provided in the present application.

The present inventors separated the holoclone (Hoclone) from human fibroblasts (Donor 12 years, Fore skin, passage 3) pre-sale from the Dept. of Dermatology through the following method.

The inventors seeded the cells in a 96 well plate to be 0.5 cells (1-2 or not / well) per well, and then cultured for 8 days without replacing the culture medium. DMEM + 10% FBS was used as a medium for culture. . The inventors observed wells with cell clusters formed after 8 days.

The present inventors treated cells with 0.25M Trypsin-EDTA for each well that formed the cell cluster, transferred them to 24 well plates for each cell lot, and proliferated the cells for 4 to 6 days. .

(2) Skin stem cell marker confirmation

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

The inventors incubated the Holoclone in a 25T Flask, and treated several compositions provided by the present specification to confirm the proliferation effect.

3-2: Confirmation of the degree of proliferation of skin stem cells by the composition containing ferrous sulfate and the composition containing alpha-lipoic acid

The present inventors seeded the same homolocolon (Hoclone) separated from human fibroblasts (Human Primary Fibroblast) (Donor 12 years, Fore skin, passage 3) in a 96 well plate at 4000 / well and DMEM + 10% FBS , Cultured for 1 day in a culture medium with 1% penicillin streptomycin.

After washing the cultured holoclone twice with PBS, the composition containing ferrous sulfate in the modified medium and the composition containing alpha-Lipoic acid in the modified medium are 100ul in the well. Each was divided equally and cultured for 72 hours to confirm the proliferation effect.

The proliferation effect of the Holoclone is through CCK-8 analysis (CCK8-Assay) common to those skilled in the art. Confirmed.

When compared to the case where 10% FBS + modified medium (hereinafter, a positive control) (FBS condition in FIG. 6) was treated, when the composition containing only ferrous sulfate is processed in the modified medium, a hololone (Holoclone) No proliferative effect was observed (see Figure 6).

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

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

(One) Holoclone proliferation effect confirmation

The present inventors seeded the Holoclone separated from human fibroblasts (Horman Primary Fibroblast) (Donor 12 years, Fore skin, passage 3) in a 96 well plate equal to 4000 / well and DMEM + 10% FBS , Cultured for 1 day in a culture medium with 1% penicillin streptomycin.

After washing the cultured Holoclone twice with PBS, the above-described composition of Experimental Example (1) to Experimental Example (9) was equally dispensed into wells in 100ul increments and cultured for 72 hours, thereby proliferating. Confirmed.

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

The present inventors show that the proliferation effect of the Holoclone by the Experimental Example (4) and Experimental Example (7) is 2.5 times higher than that of the negative control group, and the Experimental Example (5), Experimental Example (6), and Experimental Example (8) and Experimental Example (9) confirmed that the proliferation effect of the Holocalone (Holoclone) is about 3.5 times higher than that of the negative control (see FIG. 8).

Moreover, it was confirmed that the proliferation effect of the Holoclone by the 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. Did.

(2) Comparison of Holoclone Proliferation Effect and Human Primary Fibroblast Proliferation Effect

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

When the Experimental Example (8) was treated with human fibroblasts (Human Primary Fibroblast), it showed a 1.88-fold proliferative effect compared to 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, a proliferation effect of 2.99 times was observed compared to the negative control. (See Figure 9)

The present inventors confirmed that the proliferative effect of human primary fibroblasts is also shown by the above Experimental Example (8), but the proliferative effect of the Holoclone is about 2 times higher.

3-4: Whether adult stem cells proliferate from various sources

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

The present inventors conducted an experiment of treating the composition with bone marrow-derived stem cells and adipose-derived stem cells in order to confirm that the composition specifically affects skin stem cells.

The bone marrow-derived mesenchymal stem cells (Donor: Male 65 years) and fat-derived mesenchymal stem cells (Donor: Male 47 years) used as the adult stem cells were all purchased and used by Promo cells.

The present inventors cultivated the bone marrow-derived stem cells and adipose-derived stem cells by treating the experimental examples (1) to (9). In addition, the present inventors treated the modified medium (O in FIG. 10 and 0 in FIG. 11) as the negative control to the bone marrow-derived stem cells and fat-derived stem cells.

The inventors seeded bone marrow-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells in 96 well plates equally to 4000 / well and cultured for 1 day in a culture medium with MEMα medium + 10% FBS, 0.5% Gentamicin. After washing the cultured stem cells twice with PBS, the experimental examples (1) to (9) containing ferrous sulfate and lipoic acid were equally dispensed into wells for 100 ul each for 72 hours. After culturing for a while, the proliferation effect of the bone marrow-derived mesenchymal stem cells and fat-derived mesenchymal stem cells was confirmed.

When the proliferation effect was analyzed through CCK-8 analysis (CCK8-Assay), the proliferation 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] Wound healing effect confirmed by a composition containing iron sulfate (Ferrous sulfate) and alpha-lipoic acid (α-Lipoic acid)

The present inventors conducted the following experiment to confirm whether the wound healing effect by the Mix II containing ferrous sulfate and alpha-lipoic acid appeared, and as a comparison group, the Mix I composition and The MixIII composition, the modified medium (negative control) and 10% FBS + the modified medium (positive control) were used.

The present inventors scratched human fibroblasts (Human Primary Fibroblast) and treated the composition to confirm the effect of wound healing after 48 hours.

N.Cont in FIG. 12 represents the negative control, and P.Cont in FIG. 12 represents the positive control. According to Figure 12, when treated with MixⅡ-low, the wound healing (Wound Healing) effect was shown as a positive control condition.

[Example 5] Cell aging marker expression inhibition confirmed by a composition containing ferrous sulfate and alpha-lipoic acid

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

The present inventors seeded 20000 / cm 2 Holoclone at 60 ° dish and incubated for 1 day in DMEM + 10% FBS, 1% P / S.

After washing the cultured cells twice with PBS, DMEM Phenol red-free media was filled with 0.02 ml / cm 2 and exposed to UVA 6J using a UV irradiator (BLX-254, VILBER Lourmat, France), followed by Western blot (Western blot) to confirm the expression of aging markers (P21, p-P53) in the cells. (UVA 6J, C condition in FIG. 13).

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

The present inventors confirmed that the cells exposed to the UVA 6J were incubated for 72 hours in the Experimental Example (8), and then the expression levels of the P21 and the p-P53 were reduced as much as before exposure to the UVA 6J (FIG. 13). UVA 6J, Experimental Example (8) conditions)

[Example 6] Confirmation of the whitening effect by the 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 alpha-lipoic acid.

The present inventors treated α-MSH to induce melanin pigment synthesis in mouse melanocyte (B16-F1) pre-sent from Kyung Hee University, and compared with α-MSH without α-MSH. When the treatment 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 reference (1.00 value), and when α-MSH was not treated ([Table 5] negative control) 1), when the experimental example (8) was treated, the relative melanin values when the comparative examples were treated were described.

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

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 2.

Through this, the present inventors have confirmed that the whitening effect is better than when treating alpha-lipoic acid alone when combining ferrous sulfate and alpha-lipoic acid.

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 cosmetic composition containing ferrous sulfate and alpha-lipoic acid

The present inventors prepared a cosmetic composition having the composition shown in Table 6 below.

The manufacturing method of the cosmetic composition is as follows.

One) 2 to 7 (hereinafter, A) and 8 to 10 (hereinafter, B) of the following table are heated to 75 ° C, respectively.

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

3) After adding 1 and 11 of the following table, the mixture was stirred for 2 minutes.

4) Stir at 2500 rpm for 1 minute to degas the composition.

5) The beaker containing the composition is immersed in ice water and cooled to 28-30 ° C.

6) Stabilized by leaving the prepared composition at room temperature for 24 hours.

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 Monostearate sorbitan 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 Proper

[Example 8] Human fibroblast proliferation effect by a composition containing various components that can be used for cell culture

The present inventors have confirmed that the cell proliferation effect can be exhibited by other components (hereinafter referred to as "cell culture components") that can be used for cell culture in general, not ferrous sulfate and alpha-lipoic acid. The following experiment was conducted to confirm whether or not.

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

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 sulfate heptahydrate (Zinc Sulfate Heptahydrate), Riboflavin, which were purchased from Sigma Aldrich. The DMEM basic medium is Hyclone? Purchased from.

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

When compared to 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 only for illustrating the contents disclosed by the present specification, and it is understood by those skilled in the art that the scope of the contents disclosed by the present specification is not to be construed as limited by these examples. It will be obvious to you.

Claims (31)

Ferrous sulfate; and
Lipoic acid or a salt thereof;
Skin cell activation composition comprising a.
According to claim 1,
The ferrous sulfate is ferrous sulfate (iron sulfate (II); FeSO ₄ ) is a composition for activating skin cells.
According to claim 2,
The ferrous sulfate (iron (II) sulfate; FeSO ₄ ) is anhydrous salt (FeSO ₄ ), tetrahydrate (FeSO ₄ · 4H ₂ O), pentahydrate (FeSO ₄ · 5H ₂ O), hexahydrate (FeSO ₄ A composition for activating skin cells comprising any one or more of 6H ₂ O) and 7 beards (FeSO ₄ · 7H ₂ O).
According to claim 1,
The lipoic acid (Lipoic acid) is alpha lipoic acid (α-Lipoic acid) or beta lipoic acid (β-Lipoic acid) composition for activating skin cells.
According to claim 4,
The alpha-lipoic acid is (R)-(+)-alpha-lipoic acid ((R)-(+)-α-lipoic acid), (S)-(-)-alphalipoic acid ((S)- (-)-α-lipoic acid) or a mixture thereof, a composition for activating skin cells.
According to claim 1,
The ferrous sulfate is 0.5 μM to 500 μM;
The lipoic acid (Lipoic acid) is 0.5μM to 500μM skin cell activation composition.
According to claim 1,
The ferrous sulfate is 0.5 μM to 200 μM;
The lipoic acid (Lipoic acid) is 100μM to 500μM skin cell activation composition.
According to claim 1,
The ferrous sulfate is 0.5 μM to 200 μM;
The lipoic acid (Lipoic acid) is a skin cell activation composition of 200μM to 500μM.
According to claim 1,
The ferrous sulfate is 0.5 μM to 60 μM;
The lipoic acid (Lipoic acid) is a skin cell activation composition of 200μM to 400μM. According to claim 1,
The composition for activating skin cells containing the ferrous sulfate and the lipoic acid in a molar ratio of 1: 1 to 1:10.
According to claim 1,
The ferrous sulfate and the lipoic acid
Skin cell activation composition contained in a molar concentration ratio of 1: 4 to 1: 8.
According to claim 1,
The skin is dermal (Derma) or epidermis (Epiderma) skin cell activation composition.
According to claim 1,
The cells are fully differentiated cells. A composition for activating skin cells that is a progenitor cell or a stem cell.
According to claim 1,
Amino acids, sugars, vitamins, inorganic salts, lipids, cytokines, growth factors, extracellular matrix proteins, media, antioxidants, melanin pigment synthesis inhibitors, toxicity acting on cells containing melanin pigments A composition for activating skin cells further comprising at least one of a substance and an exfoliating substance.
According to claim 1,
The composition for skin cell activation, characterized in that it does not contain animal-derived serum.
The method of claim 15,
The animal-derived serum is FBS (Fetal Bovine Serum) is a composition for activating skin cells.
According to claim 1,
The composition is a composition for activating skin cells having one or more of the following functions:
i) cell proliferation effect function;
ii) cell regeneration function;
iii) cell aging regulator expression suppression function; And
iv) Whitening function.
The method of claim 17,
The aging regulator is a composition for skin cell activation, characterized in that a CDK (Cyclic-dependent Kinase) inhibitor.
The method of claim 18,
The CDK (Cyclic-dependent Kinase) inhibitor P21, P27, P57, P16, P15, P18, P19, P53 and p-P53 one or more of the composition for activating skin cells, characterized in that.
According to claim 1,
The composition is a skin cell activation composition, characterized in that in the form of a medium for cell culture.
The method of claim 20,
The medium for the cell culture is glycine (Glycine), arginine (L-Arginine), glutamine (L-Glutamine), glutamic acid (L-Glutamic Acid), asparagine (L-Asparagine-H2O), alanine ( L-Alanine, L-Aspartic acid, L-Cysteine hydrochloride-H2O, L-Histidine, L-Isoleucine, L-Leucine, Lysine ( L-Lysine, L-Methionine, L-Phenylalanine, L-Serine, L-Threonine, Tryptophan, L-Proline, Valine -Valine, Calcium pantothenate, Folic Acid, Niacinamide, Pyridoxine hydrochloride, Riboflavin, Thiamine hydrochloride, Cy Ancobalamine (Vitamin B12), Ascorbic Acid, Biotin, Inositol, Calcium Chloride (CaCl 2)), Magnesium Sulfate (MgSO4), Potassium Chloride (KCl), Sodium Bicarbonate (NaHCO3), Sodium Chloride (NaCl), Sodium Phosphate Monobasic (NaH2PO4-H2O)), glucose (D-Glucose (Dextrose)), sodium pyruvate (Sodium Pyruvate), adenosine (Adenosine) and guanosine (Guanosine) at least one composition for activation.
Ferrous sulfate; and
Lipoic acid or a salt thereof;
Cosmetic composition for activating skin cells comprising.
The method of claim 22,
The skin is a dermal (Derma) or epidermis (Epiderma) skin cell activation cosmetic composition.
The method of claim 22,
The cells are fully differentiated cells. A cosmetic composition for activating skin cells that is a progenitor cell or a stem cell.
The method of claim 22,
The cosmetic composition for skin cell activation, characterized in that it does not contain animal-derived serum.
The method of claim 22,
The cosmetic composition for activating skin cells containing the ferrous sulfate and the lipoic acid in a molar ratio of 1: 1 to 1:10.
The method of claim 22,
The cosmetic composition for activating skin cells containing the ferrous sulfate and the lipoic acid in a molar ratio of 1: 4 to 1: 8.
The method of claim 22,
The cosmetics include softening lotion, nourishing lotion, lotion, skin, body lotion, nourishing cream, massage cream, moisture cream, hand cream, eye cream, essence, cleansing foam, cleansing cream, cleansing water, pack, gel, patch, lipstick, Skin characterized by having a formulation selected from the group consisting of makeup bases, powders, foundations, body cleansers, toothpastes, mouthwashes, shampoos, rinses, hair tonics, gels, mousses, hair essences, hairdressers, and hair dyes. Cosmetic composition for cell activation.
The method of claim 22,
The composition is a cosmetic composition for activating skin cells exhibiting one or more of the following effects:
i) anti-aging effect;
ii) regenerative effect;
iii) wrinkle improvement effect;
iv) whitening effect; And
v) Skin elasticity increasing effect.
The method of claim 22,
Glycine, L-Arginine, L-Glutamine, L-Glutamic Acid, L-Asparagine-H2O, L-Alanine, Aspartic L-Aspartic acid, L-Cysteine hydrochloride-H2O, L-Histidine, L-Isoleucine, L-Leucine, L-Lysine, Methionine ( L-Methionine, L-Phenylalanine, L-Serine, L-Threonine, L-Tryptophan, Proline (L-Proline), Valine (L-Valine), Calcium Pantote D-Calcium pantothenate, Folic Acid, Niacinamide, Pyridoxine hydrochloride, Riboflavin, Thiamine hydrochloride, and Vitamin B12 , Ascorbic Acid, Biotin, Inositol, Calcium Chloride (CaCl2), Magnesium Sulfate (Magnesium S) ulfate (MgSO4), potassium chloride (Potassium Chloride (KCl)), sodium bicarbonate (NaHCO3), sodium chloride (NaCl)), sodium phosphate (Sodium Phosphate monobasic (NaH2PO4-H2O)), Glucose (D-Glucose (Dextrose)), sodium pyruvate (Sodium Pyruvate), adenosine (Adenosine) and guanosine (Guanosine) one or more of the cosmetic composition for activating skin cells.
The method of claim 22,
Animal oil, vegetable oil, wax, paraffin, starch, trakant, cellulose derivative, 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-butyl 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 derivatives, methyltaurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine , Fatty alcohol, fatty acid glyceride, Fatty acid diethanolamide, lanolin derivatives, ethoxylated glycerol fatty acid esters, stearic acid, cetostearyl alcohol, monostearic acid glycerin, squalane, concentrated glycerin, hyaluronic acid extract and sepigel 350 for skin cell activation Cosmetic composition.

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