KR20210150799A - Cosmetic composition having antibacterial effect including immune cell conditioned media - Google Patents

Abstract

The present invention relates to an antibacterial composition containing an immune cell culture medium as an active ingredient. Also, the antibacterial composition according to the present invention increases an antibacterial effect when additionally contains a natural product.

Description

Cosmetic composition having antibacterial effect including immune cell culture solution {COSMETIC COMPOSITION HAVING ANTIBACTERIAL EFFECT INCLUDING IMMUNE CELL CONDITIONED MEDIA}

The present invention relates to a cosmetic composition derived from an immune cell culture medium having an anti-bacterial effect.

Recently, as the risk of infectious bacteria of various skin diseases has emerged, antibacterial activity against bacterial infection is recognized as an important factor for maintaining healthy skin.

Therefore, Bacillus subtilis ( Bacillus subtilis ), E. coli ( Escherichia coli ), Staphylococcus aureus ( Staphylococcus aureus ), Candida albicans ( Candida albicans ), Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) Inducing various skin diseases It is meaningful to study materials with antibacterial effects that can prevent infectious diseases caused by many types of microorganisms floating in the air as well as sexual microorganisms.

In addition, as personal hygiene has emerged in recent years, the demand for antibacterial products that can be conveniently carried is gradually increasing.

Therefore, the present inventors intend to develop a composition effective for antibacterial while including an immune cell culture medium as an active ingredient.

Registered Patent 10-2058623 Registered Patent 10-1962641

Accordingly, the present inventors confirmed that the composition containing the immune cell culture medium exhibits an antibacterial effect.

Accordingly, it is an object of the present invention to provide a skin antibacterial composition comprising an immune cell culture medium as an active ingredient.

Another object of the present invention is to provide a cosmetic in the form of a pad or sheet impregnated with the composition according to the present invention.

In order to achieve the above object, in the present invention, using a solid flat plate containing various bacteria that can be grown, such as E. coli and B. subtilis, in a gel having a constant density, The same amount of control medium and immune cell culture medium were treated on the gel in which the bacteria were distributed at a constant density, and then reacted under appropriate culture conditions for a predetermined time. It was confirmed that the antibacterial particles or antibacterial substances contained in the treated culture were diffused into the gel and acted on the growing bacteria to kill them. It was confirmed that the increase in the radius increased according to the antibacterial effect of the treated material (Example 2).

Accordingly, the present invention provides a skin antibacterial composition comprising an immune cell culture medium as an active ingredient.

In one embodiment of the present invention, the composition is myricetin, scutellarein, chrysin, silymarin, icaritin, curcumin, baicalein) , hyperoside, naringin, naringenin, amentoflavone, galangin, daidzin, soporicoside and isoquercetin ) It was confirmed that the antibacterial effect increases when one or more natural products selected from the group consisting of are additionally included.

It is well known that these natural products increase the antibacterial effect. For example, antibacterial activity of flavonoids and their structure-activity relationship: An update review , Milad Iranshahy et al., DOI: 10.1002/ptr.6208), In vitro effects on biofilm viability and antibacterial and antiadherent activities of silymarin , Ebru Evren et al., DOI 10.1007/s12223-015-0399-6), Synergistic antibacterial effect of curcumin against methicillin- resistant Staphylococcus aureus , Dong-Yeul Kwon et al., Phytomedicine 20 (2013) 714-718, A Review on Antibacterial, Antiviral, and Antifungal Activity of Curcumin , Soheil Zorofchian Moghadamtousi et al., Volume 2014, Article ID 186864, 12 pages ), Cytotoxicity and antibacterial activity of scutellarein and carajurone-enriched fraction obtained from the hydroethanolic extract of the leaves of Fridericia chica (Bonpl.) LG Lohmann , Ivana MP Violante et al., ISSN: 1478-6419 (Print) 1478-6427), Synthetic derivatives of chrysin and their biological activities , Houlv Wu et al., DOI 10.1007/s00044-013-0711-4), Antibacterial activity of naringin derivatives against pathogenic strains , G. Ce´liz et al., ISSN 1364-50 72).

In one embodiment, it was confirmed that the antibacterial effect was increased when paroxetine, a natural product, was added to the antibacterial composition (Example 3).

In the present invention, "antibacterial" refers to any action that inhibits bacteria or mold, etc., and may mean suppression or removal of bacteria or fungi harmful to the human body.

In an embodiment of the present invention, when the composition comprising the immune cell culture medium according to the present invention is applied as an active ingredient, Bacillus subtilis , Escherichia coli , Staphylococcus aureus ), Candida albicans , Pseudomonas aeruginosa , Enterococcus faecalis , Klebsiella pneumoniae , Aspergillus niger , Listeria monocytogenes ( Listeria monocytogenes ), Staphylococcus epidermidis ( Staphylococcus epidermidis ) and Micrococcus luteus ( Micrococcus luteus ) Antibacterial activity against one or more dermatophytes selected from the group consisting of Confirmed.

In the present invention, "skin flora" refers to microorganisms present on the skin (resident skin flora).

In the present invention, “culture solution” means a thing obtained by culturing the immune cells of the present invention in a known liquid medium or solid medium, and in the present invention, the concept includes immune cells.

The immune cells may be natural killer cells, T cells, B cells, dendritic cells or macrophages, and the origin of immune cells is not limited.

When the composition of the present invention is a cosmetic, the cosmetic prepared by including the immune cell culture medium as an active ingredient can be prepared in the form of a general emulsified formulation and a solubilized formulation. For example, lotions such as flexible lotions or nourishing lotions, emulsions such as facial lotions and body lotions, creams such as nourishing creams, moisture creams, and eye creams, essences, cosmetic ointments, sprays, gels, packs, sunscreens, makeup bases, liquids It may be prepared in formulations such as foundation, powder, cleansing cream, cleansing lotion, makeup remover such as cleansing oil, cleansing foam, soap, body wash, etc., such as, but not limited thereto .

In addition, the cosmetic may be manufactured as a cosmetic. For example, the composition may be a cosmetic in the form of a pad or sheet impregnated with the formulation.

The cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers to be formulated in general skin cosmetics, and common ingredients include, for example, oil, water, surfactant, humectant, lower alcohol, and thickener. , a chelating agent, a colorant, a preservative, a fragrance, etc. may be appropriately blended. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.

When the composition of the present invention is for external application to the skin, it may be prepared in the form of an ointment, patch, gel, cream or spray, but is not limited thereto.

The external preparation for skin of the present invention may further include a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, water, an ionic or Nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other commonly used external preparations for the skin It may contain adjuvants commonly used in the field of dermatology, such as ingredients. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.

The composition of the present invention can provide a desirable antibacterial effect when it contains an effective amount of an immune cell culture medium. In the present invention, the term 'effective amount' refers to the amount of an immune cell culture medium capable of inhibiting, alleviating, or removing bacterial activity. The effective amount of the immune cell culture medium included in the composition of the present invention will vary depending on the form in which the composition is commercialized, the method in which the immune cell culture medium is applied to the skin, and the time it stays on the skin. Although not limited thereto, in one embodiment of the present invention, the immune cell culture medium may be included in an amount of 0.001 to 50% by weight based on the total weight of the composition.

In addition, the composition of the present invention may be used as a transdermal administration method such as directly applied to the skin or sprayed, and the administration route of the composition of the present invention may be administered through any general route as long as it can reach the target tissue.

The usage amount of the composition of the present invention may be appropriately adjusted according to individual differences or formulations such as age and the degree of lesion, and may be used for one week to several months by applying an appropriate amount to the skin once to several times a day.

Advantages and features of the present invention, and methods for achieving them, will become apparent with reference to the embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in a variety of different forms, and only these embodiments allow the disclosure of the present invention to be complete, and common knowledge in the technical field to which the present invention belongs It is provided to fully inform the possessor of the scope of the invention, and the present invention is only defined by the scope of the claims.

The composition comprising the immune cell culture medium according to the present invention as an active ingredient may exhibit an antibacterial effect.

1 shows the cytokine array results of the immune cell culture medium (conditioned media) of the present invention.
Figure 2 shows an antibacterial effect test against Escherichia coli using the immune cell culture medium of the present invention.
Figure 3 shows the antibacterial effect test for E. coli ( Escherichia coli ) and Bacillus subtilis ( Bacillus subtilis ) using the immune cell culture medium of the present invention.
4 shows the experimental results of increasing the antibacterial effect when a natural product is added to the composition according to the present invention.

Hereinafter, the present invention will be described in detail through examples. The following examples only illustrate the present invention, but the scope of the present invention is not limited to the following examples.

[Example]

Example 1: Cytokine array

30 mL to 60 mL of human peripheral blood was collected using a 10 mL vacuum collection tube treated with heparin. 15mL of Ficoll was placed in a 50mL Leucosep tube, centrifuged, and the Ficoll solution was settling down the glass membrane in the tube. The collected blood was transferred to a Leucosep, and the red blood cells and granulocyte layers were separated by centrifugation, and the monocyte layer and platelets were separated by separating them from the top. After centrifugation, separately collected lymphocytes were centrifuged to remove the supernatant. Wash the lymphocytes from which the supernatant has been removed by suspending them with PBS or NS. The number of cells in the suspension was counted using a hemocytometer, and only lymphocytes were collected by centrifugation again.

The collected lymphocytes were inoculated into 25T flak according to the appropriate concentration, and treated with RPMI medium, nutrients, and appropriate stimuli and cultured. After harvesting all of the immune cells and the immune cell culture medium cultured for about 7 days, centrifugation was performed, and the supernatant was collected and subjected to cytokine array. Using RayBio C-Series Human Cytokine Antibody Array C5, each cytokine array membrane was blocked with blocking buffer at room temperature for 30 minutes, and then reacted at 4°C overnight. After washing, the reaction was performed overnight at 4°C using HRP-streptavidin solution. After the reaction was completed, the membrane was washed, and the detection mixture buffer was reacted at room temperature for 2 minutes, followed by measurement using Davinch-chemidoc (FIG. 1).

Example 2: Using immune cell culture medium E. coli and B. subtilis antibacterial effect on

An experiment was conducted using a solid plate dish containing various viable bacteria such as E. coli and B. subtilis in a gel having a constant density. The same amount of control medium and immune cell culture medium were treated on the gel in which the bacteria were distributed at a constant density, and then reacted under appropriate culture conditions for a certain period of time. It was confirmed that the antibacterial particles or antibacterial substances contained in the treated culture were diffused into the gel and acted on the growing bacteria to kill them. It was confirmed that the increase in the radius increased according to the antibacterial effect of the treated material ( FIGS. 2 and 3 ).

Experiments were conducted using a TSA (Tryptic soy agar) solid medium in which various bacteria can grow. The immune cell culture medium was obtained according to the immune cell culture medium harvesting method, and the same amount of the control medium and the immune cell culture medium was absorbed into the TSA solid medium. The same E. coli was smeared on the absorbed TSA solid medium, and cultured overnight at _{37° C. CO 2 in an incubator.} Antibacterial activity was confirmed by counting the number of E. coli colonies cultured in each TSA solid medium in which the control medium and immune cell culture medium were absorbed ( FIGS. 2 and 3 ).

Example 3: Antibacterial activity test according to the addition of natural products

Experiments were conducted using a TSA solid medium in which various bacteria can grow. Immune cell culture medium was obtained according to the harvesting method of immune cell culture medium, and the same amount of control medium and immune cell culture medium was absorbed into TSA solid medium. In addition, the test is carried out including natural products. Myricetin having a concentration range of 0.5mM to 25mM was dispensed and absorbed in the same amount. The same E. coli was smeared on the absorbed TSA solid medium, and cultured overnight at _{37° C. CO 2 in an incubator.} By counting the number of E. coli colonies cultured in each TSA solid medium in which the control medium and the immune cell culture medium were absorbed, the antibacterial activity of the natural product was confirmed (FIG. 4).

Claims (9)

A skin antibacterial composition comprising an immune cell culture medium as an active ingredient. According to claim 1,
The composition is myricetin, scutellarein, chrysin, silymarin, icaritin, curcumin, baicalein, hyperoside. , selected from the group consisting of naringin, naringenin, amentoflavone, galangin, daidzin, soporicoside and isoquercetin The composition further comprising one or more natural products. According to claim 1,
The composition is Bacillus subtilis ( Bacillus subtilis ), E. coli ( Escherichia coli ), Staphylococcus aureus ( Staphylococcus aureus ), Candida albicans ( Candida albicans ), Pseudomonas aeruginosa ( Pseudomonas aeruginosa ), Faecalis ( Enterococcus faecalis ), Klebsiella pneumoniae ( Klebsiella pneumonia ), Aspergillus niger ( Aspergillus niger ), Listeria monocytogenes ( Listeria monocytogenes ), Staphylococcus epidermidis ( Staphylococcus epidermidis ) and Micrococcus luteus ( Micrococcus luteus ) A composition for one or more skin flora selected from the group consisting of. According to claim 1,
The composition is a cosmetic, composition. 5. The method of claim 4,
A composition in the form of a lotion, essence, lotion, cream, pack, gel or detergent. According to claim 1,
The composition is a skin external preparation, composition. 7. The method of claim 6,
A composition in the form of an ointment, patch, gel, cream or spray. According to claim 1,
The composition of the immune cell culture medium is included in an amount of 0.001 to 50% by weight based on the total weight of the composition. A cosmetic in which the composition according to claim 1 is impregnated into a pad or sheet.

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