WO2023075165A1 - Antimicrobial composition comprising antiseptic compound and reuterin, and use thereof - Google Patents

Abstract

The present invention relates to an antimicrobial composition, a cosmetic composition, and a preservative for preserving cosmetics, the antimicrobial composition comprising an antiseptic compound and reuterin in order to inhibit microbial activity or kill microorganisms. The antimicrobial composition comprises the antiseptic compound and reuterin and thus may be used as a cosmetic composition, a preservative for preserving cosmetics, etc. whereby the amount of use of the antiseptic compound is minimized, and thus formulation stability may be improved, and the amount of use may be reduced compared to when using reuterin alone, and thus antimicrobial activity in the formulation may be secured, and thus disadvantages of the antiseptic compound or reuterin are compensated for.

Description

Antimicrobial composition containing antiseptic compound and luterin and use thereof

The present invention relates to an antibacterial composition, a cosmetic composition, and a preservative for cosmetic preservation containing a preservative compound and luterin for inhibiting the activity of microorganisms or killing microorganisms.

Microorganisms decompose organic matter or organic compounds to produce substances useful to humans, but also produce substances harmful to humans with odors and chemically and biologically degenerate them. In particular, in the case of cosmetics, contamination by microorganisms during the manufacturing process and microbial contamination by skin contact such as fingers during use cannot be avoided. The use of preservatives is unavoidable.

Preservatives are formulated into cosmetics to suppress the proliferation of microorganisms that contaminate cosmetics from the outside, and to prevent product changes by killing microorganisms over time. Preservatives that are mainly used are synthetic preservatives containing chemical substances, such as paraoxybenzoic acid esters (parabens), imidazolidinyl urea, phenoxyethanol, etc., and diols with two hydroxyl groups. A based compound or surfactant may also be used as a preservative. However, among synthetic preservatives, parabens have problems such as allergic reactions, potential as endocrine disruptors, and induction of resistant bacteria, and diol-based compounds and surfactants have weak antifungal activity and dilute the product, causing problems with formulation stability and skin irritation. It is becoming. As the safety and harmfulness of these synthetic preservatives are known, consumer anxiety is increasing. Efforts have been made to reduce the amount of synthetic preservatives used, but if preservatives are not added or added at a level that is less than the appropriate amount, the shelf life of the product is shortened and product deterioration and microbial contamination become problems. The reality is that you have no choice but to use .

In order to overcome the problems of these synthetic preservatives, studies using materials obtained from nature or developing natural preservatives containing them are being actively conducted. For example, there has been an attempt to use extracts extracted from plants such as anise, portulaca, centella asiatica, ginkgo biloba, quince leaf, and horsetail leaf by mixing them with synthetic preservatives. However, there is still a need for research and development of preservatives capable of securing both product performance and stability while reducing the amount of use compared to conventional synthetic preservatives.

On the other hand, luterin, a natural antibacterial substance, has been reported as a substance showing a wide spectrum of antimicrobial activity. However, since the antibacterial activity is low in the presence of amino acids, it is difficult to apply to products containing amino acids (Eunmi Yeom et al., Journal of Korean Food Science 2004. 36(1): 111-115).

Therefore, many studies are needed to overcome the disadvantages of existing preservatives such as synthetic preservatives and luterin.

[Prior art literature]

[Patent Literature]

(Patent Document 1) Republic of Korea Patent Registration No. 10-2013565

(Patent Document 2) Republic of Korea Patent Registration No. 10-2136623

(Patent Document 3) Republic of Korea Patent Registration No. 10-1472518

An object of the present invention is to provide an antibacterial composition for inhibiting the activity of microorganisms such as bacteria and fungi or killing microorganisms, a cosmetic composition, and a preservative for cosmetic preservation.

One aspect of the present invention provides an antimicrobial composition comprising an antiseptic compound and luterin as active ingredients.

The term "preservative compound" used in the present invention refers to a compound added to prevent chemical or biological deterioration of a substance, and may include a compound used as a preservative or disinfectant. In view of the chemical toxicity of preservative compounds, cosmetics, foods, and medicines must use safe substances that are not toxic to the human body, so in the present invention, any preservative compound that is chemically safe and harmless can be used without limitation.

Examples of the preservative compound include parabens, paraffins, alkanediols, phenoxyethanol, imidazolidinyl urea, diazolidinyl urea ), isopropyl alcohol, chlorophene, formic acid, piroctone olamine, hexetidine, hexamidine, isothiazolinone ), hydroxyacetophenone, glyceryl caprylate, ethylhexylglycerin, sodium benzoate, etc., but are not limited thereto.

According to one embodiment of the present invention, the preservative compound may be at least one selected from the group consisting of alkane diol, hydroxyacetophenone, glyceryl caprylate, ethylhexyl glycerin and sodium benzoate.

The alkane diol refers to a compound in which two hydroxyl groups (-0H) are connected to a saturated hydrocarbon (alkane) that forms a chain with a single bond between carbons. It is classified as 1,4-, 2,3-, 2,4-, 2,5-, etc. These alkanediols are mainly used as moisturizing ingredients in cosmetics and pharmaceuticals, but since they contain a hydrophilic hydroxyl group and a hydrophobic carbon chain, they are amphiphilic and can easily penetrate cell membranes of microorganisms, thereby exhibiting excellent antibacterial activity. In particular, the longer the carbon chain of the alkanediol, the stronger the hydrophobicity, so the antibacterial activity increases, but the higher the absorption rate in the body, causing skin irritation. In the present invention, the alkanediol preferably has 3 to 12 carbon atoms.

According to one embodiment of the present invention, the alkane diol may be an alkane diol having 3 to 12 carbon atoms.

More specifically, the alkanediol includes propanediol, butanediol, pentanediol, hexanediol, octanediol, decanediol and dodecanediol. It may be one or more selected from the group consisting of.

According to one embodiment of the present invention, the alkanediol may be at least one selected from the group consisting of propanediol, butanediol, pentanediol, hexanediol, and octanediol.

In particular, since hydrophilicity improves as hydroxyl groups of alkanediol are concentrated, 1,2-alkanediol, 1,3-alkanediol, or 2,3-alkanediol is preferable to enhance antibacterial activity. For example, 1,2-propanediol, 1,3-propanediol, 1,2-butanediol, 1,3-butanediol, 2,3-butanediol, 1,2-pentanediol, 1,3-pentanediol, 2,3-pentanediol, 1,2-hexanediol, 1,3-hexanediol, 2,3-hexanediol, 1,2-octanediol, 1,3-octanediol, 2,3-octanediol, etc. However, it is no longer limited.

The hydroxyacetophenone is a plant-derived ingredient used as a natural antioxidant in cosmetics, and has a weaker preservative power than parabens, but is more skin-friendly and increases the preservative function when used with other preservatives including parabens. These hydroxyacetophenones are classified into 2-, 3- and 4-hydroxyacetophenones according to the position of the hydroxy group (-OH), but there is no difference in preservative function.

The glyceryl caprylate is a plant-derived ingredient that serves as a kind of surfactant and is mainly used as an emulsifier in cosmetics, and has excellent antibacterial and antifungal effects, so it can be used as a substitute for preservatives.

The ethylhexylglycerin has a wetting property and is mainly used for the moisturizing effect of cosmetics, but also serves to increase the antibacterial power of common preservatives such as phenoxyethanol, isothiazolinone, and paraben, and is used to reduce the amount of such preservatives. It also has antibacterial activity that inhibits the growth of odor-causing anaerobic bacteria in the human body.

The sodium benzoate is a compound that is effective in inhibiting the growth of various microorganisms such as bacteria, yeast, and fungi. It is inexpensive, has excellent antibacterial effect, and is low in toxicity, and is mainly used as a food preservative. In cosmetics, it replaces parabens. It is one of the synthetic preservatives.

As used in the present invention, “reuterin” refers to 3-hydroxypropionaldehyde, a natural antimicrobial substance produced by Lactobacillus reuteri from glycerol. The luterin has antibacterial activity against not only pathogenic bacteria but also yeast and mold, and thus has a very broad antibacterial spectrum (Doleyres et al., Appl Microbiol Biotechnol 2005, 68: 467-474).

According to one embodiment of the present invention, the luterin may be isolated from Lactobacillus reuteri or a culture medium thereof.

The "culture medium" used in the present invention refers to nutrients necessary for the survival, development, or proliferation of Lactobacillus reuteri, and may also include enzymes and metabolites produced by the Lactobacillus reuteri and secreted to the outside.

A composition comprising such an antiseptic compound and luterin may exhibit antibacterial activity.

"Antimicrobial" as used in the present invention means the ability to reduce, prevent, inhibit or eliminate the growth or survival of microorganisms such as bacteria and fungi at a certain concentration, and "antimicrobial composition" refers to substances exhibiting such antibacterial activity. means a composition comprising

According to one embodiment of the present invention, the antimicrobial composition may have antibacterial activity against bacteria and fungi.

According to one embodiment of the present invention, the antibacterial composition is Escherichia (Escherichia ) genus, Staphylococcus ( Staphylococcus ) genus, Pseudomonas ( Pseudomonas ) genus, Candida ( Candida ) and Aspergillus ( Aspergillus ) genus It may have antibacterial activity against one or more bacteria selected from the group consisting of.

The antibacterial composition contains a preservative compound and luterin, thereby exhibiting sufficient antibacterial activity even in a small amount compared to the amount of preservative compound or luterin used for antibacterial activity in the past, maintaining formulation stability of the product and extending shelf life. can do.

According to one embodiment of the present invention, the antimicrobial composition may be one containing 0.01 to 50% by weight of an antiseptic compound based on the total weight.

More specifically, the amount of the preservative compound is 0.01 to 50% by weight, 0.01 to 10% by weight, 0.01 to 5% by weight, 0.01 to 1% by weight, 0.05 to 50% by weight, 0.05 to 50% by weight based on the total weight of the antimicrobial composition. to 10 wt%, 0.05 to 5 wt%, 0.05 to 1 wt%, 0.1 to 50 wt%, 0.1 to 10 wt%, 0.1 to 5 wt%, 0.1 to 1 wt%, 0.5 to 50 wt%, 0.5 to 10 0.5 to 5 wt%, 0.5 to 1 wt%, 1 to 50 wt%, 1 to 10 wt%, 1 to 5 wt%, 5 to 50 wt%, or 10 to 50 wt%.

When the amount of the preservative compound is less than 0.01% by weight, antibacterial activity against bacteria and fungi is not shown, and a large amount of luterin is unavoidable to improve antibacterial activity, and when the amount of the preservative compound exceeds 50% by weight, the product This thinning may cause a problem with formulation stability and greatly increase skin irritation.

Further, according to one embodiment of the present invention, the antimicrobial composition may contain 0.05 to 50 mM of luterin.

The concentration of luterin in the antimicrobial composition is 0.05 to 50 mM, 0.05 to 10 mM, 0.05 to 5 mM, 0.1 to 50 mM, 0.1 to 10 mM, 0.1 to 5 mM, 0.5 to 50 mM, 0.5 to 10 mM, 0.5 to 5 mM, 1 to 50 mM, 1 to 10 mM, or 1 to 5 mM.

If the concentration of luterin is less than 0.05 mM, it may not exhibit antibacterial activity or may show limited antibacterial activity only in some microorganisms, and if the concentration of luterin exceeds 50 mM, formulation stability may be adversely affected.

According to one embodiment of the present invention, luterin exhibits antibacterial activity against bacteria ( E. Coli , P. aeruginosa and S. aureus ) and fungi ( C. albicans and A. niger ) even when used alone, but is a preservative compound. , namely alkanediol (1,3-propanediol, 2,3-butanediol, 1,2-pentanediol, 1,2-hexanediol or 1,2-octanediol), hydroxyacetophenone, glyceryl caprylate, It was confirmed that the mixed use with ethylhexylglycerin or sodium benzoate showed the same or better antibacterial activity with only a small amount compared to the use of luterin alone.

This antibacterial composition can be applied in any formulation such as liquid, oil, cream, paste, solid, etc., and can be used for products applied to the body, such as the body, skin, oral cavity, and hair.

The antibacterial composition according to the present invention may be utilized in various fields depending on the purpose or aspect of use, and may be embodied as, for example, a cosmetic composition or an additive thereof. The additive is an additive for preventing deterioration, decay, discoloration, and chemical change of the product by microorganisms, and may be a preservative, a preservative, a bactericide, etc., based on the total weight of the cosmetic composition, 0.01 to 50% by weight, 0.5 to 25% by weight, 0.1 to 10% by weight, or 0.5 to 5% by weight may be included.

Antibacterial composition according to the present invention can be implemented as a cosmetic composition.

According to one embodiment of the present invention, the cosmetic composition may contain an antibacterial compound and luterin as active ingredients.

Since the description of the preservative compound and luterin is the same as described above, it is omitted to avoid redundant description.

In addition to the preservative compound and luterin, the cosmetic composition may further include ingredients commonly used in cosmetic compositions, for example, antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances, and the like.

The cosmetic composition may be prepared without limitation in formulations commonly used in the art, for example, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream , Moisture Cream, Hand Cream, Essence, Nutrition Essence, Pack, Soap, Shampoo, Cleansing Foam, Cleansing Lotion, Cleansing Cream, Body Lotion, Body Cleanser, Emulsion, Lipstick, Makeup Base, Foundation, Press Powder, Loose Powder, Eye Shadow It can be prepared in formulations such as

According to one embodiment of the present invention, the cosmetic composition may include 0.01 to 50% by weight of an antiseptic compound based on the total weight.

More specifically, the amount of the preservative compound is 0.01 to 50% by weight, 0.01 to 10% by weight, 0.01 to 5% by weight, 0.01 to 1% by weight, 0.05 to 50% by weight, 0.05 to 50% by weight based on the total weight of the cosmetic composition. 10 wt%, 0.05 to 5 wt%, 0.05 to 1 wt%, 0.1 to 50 wt%, 0.1 to 10 wt%, 0.1 to 5 wt%, 0.1 to 1 wt%, 0.5 to 50 wt%, 0.5 to 10 wt% %, 0.5 to 5%, 0.5 to 1%, 1 to 50%, 1 to 10%, 1 to 5%, 5 to 50%, or 10 to 50%.

In addition, according to one embodiment of the present invention, the cosmetic composition may contain 0.05 to 50 mM of luterin.

The concentration of luterin in the cosmetic composition is 0.05 to 50 mM, 0.05 to 10 mM, 0.05 to 5 mM, 0.1 to 50 mM, 0.1 to 10 mM, 0.1 to 5 mM, 0.5 to 50 mM, 0.5 to 10 mM, 0.5 to 5 mM, 1 to 50 mM, 1 to 10 mM, or 1 to 5 mM.

In addition, the antibacterial composition according to the present invention can be implemented as a preservative for preserving cosmetics.

According to one embodiment of the present invention, the preservative for preserving cosmetics may be a preservative for preserving cosmetics containing a preservative compound and luterin as active ingredients.

Since the description of the preservative compound and luterin is the same as described above, it is omitted to avoid redundant description.

The preservative for preserving cosmetics is an additive used in manufacturing cosmetics to prevent spoilage, and particularly serves to prevent biological deterioration.

According to one embodiment of the present invention, the preservative for preserving cosmetics may include 0.01 to 50% by weight of a preservative compound based on the total weight.

More specifically, the amount of the preservative compound is 0.01 to 50% by weight, 0.01 to 10% by weight, 0.01 to 5% by weight, 0.01 to 1% by weight, 0.05 to 50% by weight, based on the total weight of the preservative for cosmetic preservation. 0.05 to 10% by weight, 0.05 to 5% by weight, 0.05 to 1% by weight, 0.1 to 50% by weight, 0.1 to 10% by weight, 0.1 to 5% by weight, 0.1 to 1% by weight, 0.5 to 50% by weight, 0.5 to 10 wt%, 0.5 to 5 wt%, 0.5 to 1 wt%, 1 to 50 wt%, 1 to 10 wt%, 1 to 5 wt%, 5 to 50 wt%, or 10 to 50 wt%.

Further, according to one embodiment of the present invention, the preservative for preserving cosmetics may contain 0.05 to 50 mM of luterin.

The concentration of luterin is 0.05 to 50 mM, 0.05 to 10 mM, 0.05 to 5 mM, 0.1 to 50 mM, 0.1 to 10 mM, 0.1 to 5 mM, 0.5 to 50 mM, 0.5 to 10 mM in the preservative for cosmetic preservation. , 0.5 to 5 mM, 1 to 50 mM, 1 to 10 mM, or 1 to 5 mM.

The preservative for preserving cosmetics may further include components commonly used depending on the type and purpose of use of the product, and may further include, for example, acetic acid, sodium acetate-based, sorbic acid-based, benzoic acid-based, propionic acid-based, etc. .

The antibacterial composition according to the present invention can improve formulation stability by minimizing the amount of preservative compound and luterin by including a preservative compound and luterin, and can secure antimicrobial activity in the formulation even if the amount used is reduced compared to the use of a preservative compound or luterin alone. It can be used as a preservative compound or a cosmetic composition that compensates for the disadvantages of luterin, a preservative for cosmetic preservation, and the like.

1 is a graph showing the change in viscosity of a cosmetic composition including an antibacterial composition according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail. However, these descriptions are merely presented as examples to aid understanding of the present invention, and the scope of the present invention is not limited by these exemplary descriptions.

Preparation Example 1. Preparation of preservative compound and luterin

1-1. preservative compound

1,3-propanediol (1,3-PD), 2,3-butanediol (2,3-BD), 1,2-pentanediol (1,2-PD), 1,2-hexanediol as preservative compounds Alkanediols of (1,2-HD) and 1,2-octanediol (1,2-OD), 4-hydroxyacetophenone (4-HA), glycerylcaprylate (GC), ethylhexylglycerin (EHG) ) and sodium benzoate (SB) were used.

1-2. luterin

Reuterin was obtained and used through microbial culture.

Briefly, MRS medium (10 g/L Proteose Peptone NO.3, 10 g/L beef extract, 5 g/L yeast extract, 20 g /L dextrose, 1 g/L polysorbate 80, 2 g/L ammonium citrate, 5 g/L sodium acetate, 0.1 g/L magnesium sulfate, 0.05 g/L manganese sulfate, and After culturing in 2 g/L of dipotassium sulfate), the culture medium was centrifuged at 13,000 rpm for 5 minutes, and the cells were collected and washed twice in 50 mM sodium phosphate buffer (pH 7.5). The washed cells were resuspended in a 200 mM glycerol solution, allowed to stand in an incubator at 30° C., and samples were taken at intervals of 1 hour. Reuterin at a concentration of 100 mM was obtained and used in subsequent examples.

Example 1. Antibacterial evaluation of antibacterial composition

Antibacterial evaluation was performed on the antibacterial composition containing alkanediol and luterin.

1-1. MIC evaluation

In the antimicrobial activity test of the antibacterial composition, three types of bacteria ( Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus ATCC 6538) and two types of fungi ( Candida albicans ATCC 10231, Aspergillus niger ATCC 16404) designated by the United States Pharmacopoeia (USP) were used. MIC (minimal inhibitory concentration) method was performed. The bacteria were subcultured twice in TSB medium and the fungi in SDB medium, and then used in experiments.

First, as an antimicrobial composition for each medium (bacteria - TSB medium, fungus - SDB medium), luterin alone or luterin and alkanediol were mixed at a weight ratio of 1: 1 and added to 0.5 to 4% by weight. At this time, in the medium The final luterin concentration was 0.5 to 4 mM. Thereafter, bacteria were inoculated to a concentration of 10 ⁷ cfu/ml and cultured at 37°C for 1 day, or fungi were inoculated to a concentration of 10 ⁵ cfu/ml and cultured at 30°C for 2 days to confirm the MIC value. The results are shown in Table 1 below.

	MIC (wt%)
	E. Coli	S. Aureus	P. aeruginosa	C. albicans	A. niger
luterin	4	2	2	4	2
Reuterin+1,2-HD	2	2	2	2	2

As shown in Table 1, when luterin alone was used as a preservative, antibacterial activity was shown at 4 wt% luterin (luterin 4 mM) against E. coil and C. albicans , but luterin and 1,2-HD In the case of mixed use, the antibacterial activity was confirmed in 1 wt% of luterin (1 mM of luterin), and it was confirmed that the antibacterial activity was increased compared to the use of luterin alone.

1-2. MBC evaluation

The antibacterial composition contains 0.1 to 30 wt. weight of alkanediol (1,3-propanediol, 2,3-butanediol, 1,2-pentanediol, 1,2-hexanediol or 1,2-octanediol) in 0.85% NaCl buffer. % and/or 0.1 to 3 wt% (0.1 to 3 mM) of lutein. 10 ⁵ cfu/mL of C. albicans or A. niger was inoculated in 10 mL of the antibacterial composition and cultured with shaking for 72 hours in a 30°C incubator. Thereafter, 100 μL of the culture medium was spread on SDA solid medium and cultured for 48 hours in a 30° C. incubator to observe bacterial growth. The MBC (minimum bactericidal concentration) evaluation results for C. albicans and A. niger are shown in Tables 2 and 3, respectively.

C. albicans		Reuterin (mM)
		0	0.1	0.5	1.0	1.5	2.0	3.0
Alkanediol not added		X	X	X	X	O		O	0
1,3-PD (wt%)	One	X	X	X	O	O	O	O
	2	X	X	O	O	O	O	O
	3	X	O	O	O	O	O	O
2,3-BD (wt%)	10	X	X	X	O	O	O	O
	15	O	O	O	O	O	O	O
	20	O	O	O	O	O	O	O
	25	O	O	O	O	O	O	O
	30	O	O	O	O	O	O	O
1,2-PD (wt%)	3	X	X	X	O	O	O	O
	5	X	O	O	O	O	O	O
	8	O	O	O	O	O	O	O
1,2-HD (wt%)	1.0	X	X	X	O	O	O	O
	2.0	O	O	O	O	O	O	O
	2.5	O	O	O	O	O	O	O
	3.0	O	O	O	O	O	O	O
1,2-OD (wt%)	0.1	X	X	X	O	O	O	O
	0.3	X	X	X	O	O	O	O
	0.5	X	O	O	O	O	O	O
X: No antibacterial activity (bacterial growth), O: Antibacterial activity (no bacterial growth)

A. niger		Reuterin (mM)
		0	0.1	0.5	1.0	1.5	2.0	3.0
Alkanediol not added		X	X	X	X	O		O	0
1,3-PD (wt%)	One	X	X	X	X	O	O	O
	2	X	X	X	O	O	O	O
	3	X	O	O	O	O	O	O
2,3-BD (wt%)	10	X	X	X	O	O	O	O
	15	X	X	X	O	O	O	O
	20	X	X	O	O	O	O	O
	25	X	X	O	O	O	O	O
	30	X	O	O	O	O	O	O
1,2-PD (wt%)	3	X	X	X	O	O	O	O
	5	X	X	O	O	O	O	O
	8	X	O	O	O	O	O	O
1,2-HD (wt%)	1.0	X	X	X	O	O	O	O
	2.0	X	X	X	O	O	O	O
	2.5	X	X	O	O	O	O	O
	3.0	X	O	O	O	O	O	O
1,2-OD (wt%)	0.1	X	X	X	O	O	O	O
	0.3	X	X	X	O	O	O	O
	0.5	X	X	O	O	O	O	O
X: No antibacterial activity (bacterial growth), O: Antibacterial activity (no bacterial growth)

Referring to Tables 2 and 3, for C. albicans , 1 to 3% by weight of 1,3-propanediol, 10% by weight of 2,3-butanediol, 3 to 5% by weight of 1,2-pentanediol, 1, 1% by weight of 2-hexanediol or 0.1 to 0.5% by weight of 1,2-octanediol was found to have no antibacterial activity, but antifungal activity was confirmed when 0.1 to 3 mM of luterin was further included. In addition, for A. niger , 1 to 3% by weight of 1,3-propanediol, 10 to 30% by weight of 2,3-butanediol, 3 to 8% by weight of 1,2-pentanediol, 1,2-hexanediol ~ 3% by weight or 0.1 ~ 0.5% by weight of 1,2-octanediol did not show antibacterial activity, but it was confirmed that antibacterial activity was exhibited when 0.1 ~ 3 mM of luterin was further included. Therefore, it was found that the composition containing alkanediol and luterin had an increased antibacterial activity compared to the single material through the combination of the two materials.

Example 2. Evaluation of Formulation Stability of Cosmetic Composition

2-1. Preparation of cosmetic composition

Formulation stability was confirmed by preparing a cosmetic composition containing an alkanediol alone or an alkanediol and luterin. 1,2-HD was used as the alkanediol.

The raw materials and contents of the cosmetic composition are shown in Table 4 below, and were prepared in a lotion formulation.

raw material name	Cosmetic Composition 1	Cosmetic Composition 2
Carbopol 981	0.2wt%	0.2wt%
Glycerin	5.0wt%	5.0wt%
Cetyl alcohol	0.5wt%	0.5wt%
Stearyl alcohol	0.5wt%	0.5wt%
MCTs	2.0wt%	2.0wt%
LP 70	3.0wt%	3.0wt%
Olivem 1000	1.5wt%	1.5wt%
Triethanolamine	0.2wt%	0.2wt%
1,2-HD	3.0wt%	1.0wt%
Reuterin	0	1.0 wt % (1 mM)
Distilled water	To 100wt%	To 100wt%

2-2. Formulation stability evaluation

After storing the cosmetic compositions 1 and 2 at room temperature for 60 days, the viscosity change of the formulation was observed. Viscosity evaluation (unit: cps) was conducted by putting each cosmetic composition in a transparent container and operating a viscometer (LV pro VISCOMETER, BROOKFIELD) equipped with a No. 64 spindle at 25 ° C. at 50 rpm.

As a result, as shown in FIG. 1, cosmetic composition 1 containing 1,2-HD alone showed a decrease in viscosity, but cosmetic composition 2 containing 1,2-HD and luterin showed no change in viscosity. confirmed that

Example 3. Antibacterial evaluation of antibacterial composition

An antimicrobial composition containing hydroxyacetophenone, glyceryl caprylate, ethylhexyl glycerin or sodium benzoate, and luterin was subjected to antibacterial evaluation.

3-1. Antifungal evaluation

In the antibacterial activity test against fungi, fungi Candida albicans ATCC 10231 and Aspergillus niger ATCC 16404 designated by the United States Pharmacopoeia (USP) were used. C. albicans was subcultured twice in SDB medium, and A. niger spores obtained by plating on SDA medium were used in the experiment.

As an antibacterial composition, luterin alone or luterin and an antiseptic compound were added at respective concentrations to a 0.8% NaCl aqueous solution, and the final concentration of luterin was 0.5 to 1.5 mM. Thereafter, the fungus was inoculated to 10 ⁵ cfu/ml, stored at 30° C. for 3 days, and then 100 uL was smeared on SDA medium. The smeared samples were incubated at 30° C. for 3 days to confirm whether or not colonies were formed. The results are shown in Table 5 below.

preservative compound	Reuterin (mM)	C. albicans	A. niger
not added	0	O	O
	0.5	O	O
	One	O	O
	1.5	X	X
4-HA 0.1wt%	0	O	O
	0.5	X	X
	One	X	X
	1.5	X	X
4-HA 0.2wt%	0	O	O
	0.5	X	X
	One	X	X
	1.5	X	X
GC 0.1wt%	0	O	O
	0.5	X	O
	One	X	X
	1.5	X	X
GC 0.2wt%	0	O	O
	0.5	X	O
	One	X	X
	1.5	X	X
EHG 0.05wt%	0	O	O
	0.5	X	O
	One	X	X
	1.5	X	X
EHG 0.1wt%	0	X	O
	0.5	X	X
	One	X	X
	1.5	X	X
SB 0.3wt%	0	O	O
	0.5	X	O
	One	X	X
	1.5	X	X
SB 0.4wt%	0	O	O
	0.5	X	O
	One	X	X
	1.5	X	X
X: No antibacterial activity (bacterial growth), O: Antibacterial activity (no bacterial growth)

Referring to Table 5, for C. albicans , 0.1 to 0.2% by weight of hydroxyacetophenone, 0.1 to 0.2% by weight of glyceryl caprylate, 0.05% by weight of ethylhexylglycerin or sodium benzoate, except for 0.05% by weight of ethylhexylglycerin. When 0.3 to 0.4% by weight of ate was used alone, there was no antifungal activity, but it was confirmed that antifungal activity was exhibited when luterin was further included regardless of the concentration.

In addition, for A. niger , antifungal activity was observed when 0.1 to 0.2% by weight of hydroxyacetophenone, 0.1 to 0.2% by weight of glyceryl caprylate, 0.05 to 0.1% by weight of ethylhexylglycerin, or 0.3 to 0.4% by weight of sodium benzoate was used alone. However, it was confirmed that antifungal activity appeared as the concentration of luterin increased.

3-2. Antibacterial evaluation

In the antibacterial test against bacteria, Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 9027, and Staphylococcus aureus ATCC 6538 designated by the United States Pharmacopoeia (USP) were used. Bacteria were subcultured twice in TSB medium and used in experiments.

As an antibacterial composition, luterin alone or luterin and an antiseptic compound were added at respective concentrations to a 0.8% NaCl aqueous solution, and the final concentration of luterin was 0.5 to 1.5 mM. Thereafter, the bacteria were inoculated to a concentration of 10 ⁷ cfu/ml, stored at 37° C. for 3 days, and then 100 uL was smeared on a TSA medium. The smeared sample was incubated at 37 ° C. to confirm the presence or absence of colony formation. The results are shown in Table 6 below.

preservative compound	Reuterin (mM)	E. coli	P. aeruginosa	S. aureus
not added	0	O	O	O
	0.5	O	O	O
	One	O	O	O
	1.5	X	X	X
4-HA 0.1wt%	0	O	O	O
	0.5	X	X	X
	One	X	X	X
	1.5	X	X	X
4-HA 0.2wt%	0	O	O	O
	0.5	X	X	X
	One	X	X	X
	1.5	X	X	X
GC 0.1wt%	0	O	O	O
	0.5	X	X	X
	One	X	X	X
	1.5	X	X	X
GC 0.2wt%	0	O	O	O
	0.5	X	X	X
	One	X	X	X
	1.5	X	X	X
EHG 0.05wt%	0	O	O	O
	0.5	X	X	X
	One	X	X	X
	1.5	X	X	X
EHG 0.1wt%	0	O	O	O
	0.5	X	X	X
	One	X	X	X
	1.5	X	X	X
SB 0.3wt%	0	O	O	O
	0.5	X	X	X
	One	X	X	X
	1.5	X	X	X
SB 0.4wt%	0	O	O	O
	0.5	X	X	X
	One	X	X	X
	1.5	X	X	X
X: No antibacterial activity (bacterial growth), O: Antibacterial activity (no bacterial growth)

Referring to Table 6, when each preservative compound was used alone, there was no antibacterial activity against three types of bacteria, but when luterin was added, antibacterial activity was shown, and a lower concentration of lutein than when luterin was used alone It was confirmed that the addition of terin exerted excellent antibacterial activity.

Example 4. Formulation stability and antibacterial activity maintenance evaluation of cosmetic composition

2-1. Preparation of cosmetic composition

Cosmetics of lotion and sheet mask formulations as shown in Tables 7 and 8 below to confirm the antimicrobial activity of preservative compounds (hydroxyacetophenone, glyceryl caprylate, ethylhexylglycerin or sodium benzoate) and / or luterin in cosmetic compositions A composition was prepared.

lotion formulation
raw material name	Cosmetic Composition 3	Cosmetic Composition 4	Cosmetic Composition 5	Cosmetic Composition 6	Cosmetic Composition 7
BUTYLENE GLYCOL	5.00	5.00	5.00	5.00	5.00
GLYCERIN	4.00	4.00	4.00	4.00	4.00
CETYL ETHYLHEXANOATE	2.50	2.50	2.50	2.50	2.50
GLYCERYL STEARATE	1.78	1.78	1.78	1.78	1.78
CETEARYL ALCOHOL	1.50	1.50	1.50	1.50	1.50
CAPRYLIC/CAPRIC TRIGLYCERIDE	1.50	1.50	1.50	1.50	1.50
OLEA EUROPAEA (OLIVE) FRUIT OIL	1.00	1.00	1.00	1.00	1.00
PEG-100 STEARATE	0.72	0.72	0.72	0.72	0.72
BUTYROSPERMUM PARKII(SHEA) BUTTER	0.50	0.50	0.50	0.50	0.50
TROMETHAMINE	0.40	0.40	0.40	0.40	0.40
CARBOMER	0.40	0.40	0.40	0.40	0.40
XANTHAN GUM	0.05	0.05	0.05	0.05	0.05
DISODIUM EDTA	0.02	0.02	0.02	0.02	0.02
ALLANTOIN	0.01	0.01	0.01	0.01	0.01
HYDROXYACETOPHENONE	-	0.10	-	-	-
ETHYLHEXYLGLYCERIN	-	-	0.05	-	-
GLYCERYL CAPYLATE	-	-	-	0.2	-
SODIUM BENZOATE	-	-	-	-	0.4
REUTERIN	1 mM	1 mM	1 mM	1 mM	1 mM
Water	To 100wt%	To 100wt%	To 100wt%	To 100wt%	To 100wt%

sheet mask formulation
raw material name	cosmetic composition 8	Cosmetic Composition 9	cosmetic composition 10	cosmetic composition 11	cosmetic composition 12
BUTYLENE GLYCOL	5.05	5.05	5.05	5.05	5.05
GLYCERIN	2.00	2.00	2.00	2.00	2.00
DIETHOXYETHYL SUCCINATE	2.00	2.00	2.00	2.00	2.00
PANTHENOL	0.98	0.98	0.98	0.98	0.98
TROMETHAMINE	0.10	0.10	0.10	0.10	0.10
CARBOMER	0.10	0.10	0.10	0.10	0.10
BETAINE	0.10	0.10	0.10	0.10	0.10
SODIUM HYALURONATE	0.01	0.01	0.01	0.01	0.01
HYDROXYACETOPHENONE	-	0.10	-	-	-
ETHYLHEXYLGLYCERIN	-	-	0.05	-	-
GLYCERYL CAPYLATE	-	-	-	0.2	-
SODIUM BENZOATE	-	-	-	-	0.4
REUTERIN	1 mM	1 mM	1 mM	1 mM	1 mM
WATER	To 100wt%	To 100wt%	To 100wt%	To 100wt%	To 100wt%

4-2. Evaluation of antibacterial activity in cosmetic formulations

To evaluate the antibacterial activity in the cosmetic composition, a test was conducted using the above formulations. In the antibacterial test, bacteria and fungi designated by the United States Pharmacopoeia (USP) ( Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus ATCC 6538, Candida albicans ATCC 10231 and Aspergillus niger ATCC 16404) were used. Bacteria and C. albicans were subcultured twice in TSB and SDB medium, respectively, and used in the experiment, and A. niger spores obtained by plating on SDA medium were used in the experiment.

Each cosmetic composition was inoculated to 10 ⁷ cfu/ml for bacteria and 10 ⁵ cfu/ml for fungi, and then stored at 37°C and 30°C for 3 days, respectively. After mixing 100 uL of the sample with 900 uL of Letheen medium, it was smeared on TSA for bacteria and SDA for fungi, respectively. The smeared samples were incubated at 37 ° C and 30 ° C for 3 days, respectively, to confirm the presence or absence of colony formation. The results are shown in Table 9 below.

	E. coli	P. aeruginosa	S. aureus	C. albicans	A. niger
Cosmetic Composition 3	O	O	O	O	O
Cosmetic Composition 4	X	X	X	X	X
Cosmetic Composition 5	X	X	X	X	X
Cosmetic Composition 6	X	X	X	X	X
Cosmetic Composition 7	X	X	X	X	X
cosmetic composition 8	O	O	O	O	O
Cosmetic Composition 9	X	X	X	X	X
cosmetic composition 10	X	X	X	X	X
cosmetic composition 11	X	X	X	X	X
cosmetic composition 12	X	X	X	X	X
X: No antibacterial activity (bacterial growth), O: Antibacterial activity (no bacterial growth)

Referring to Table 9, regardless of the formulation, it was confirmed that the cosmetic composition containing the preservative compound and luterin had antibacterial activity against bacteria and fungi without changing the formulation.

4-3. Evaluation of maintenance of antibacterial activity in cosmetic formulations

In order to check whether the antibacterial activity in the cosmetic formulation is maintained, '4-2. It was confirmed whether the antibacterial activity was maintained by cumulatively inoculating each strain using the bacterial or fungal inoculated samples (cosmetic compositions 4 to 7 and 9 to 12) used in 'Evaluation of antibacterial activity in cosmetic formulations'. In the case of bacteria, three species, namely E. coli , P. aeruginosa and S. aureus were mixed and inoculated to 10 ⁷ CFU/ml, and in the case of fungi, C. albicans and A. niger were mixed to 10 ⁵ CFU/ml, respectively. Inoculated. Inoculation of each strain was inoculated into the same sample every 7 days, and for the sample after inoculation, 100 uL of the sample was mixed with 900 uL of Letheen medium on the 3rd day of each inoculation, and then smeared on TSA and SDA, respectively, to check for colon formation. Confirmed. The results are shown in Table 10 below.

		Cosmetic Composition 4	Cosmetic Composition 5	Cosmetic Composition 6	Cosmetic Composition 7	Cosmetic Composition 9	cosmetic composition 10	cosmetic composition 11	cosmetic composition 12
1st inoculation	3 types of bacteria	X	X	X	X	X	X	X	X
	C. albicans	X	X	X	X	X	X	X	X
	A. niger	X	X	X	X	X	X	X	X
2nd dose	3 types of bacteria	X	X	X	X	X	X	X	X
	C. albicans	X	X	X	X	X	X	X	X
	A. niger	X	X	X	X	X	X	X	X
3rd dose	3 types of bacteria	X	X	X	X	X	X	X	X
	C. albicans	X	X	X	X	X	X	X	X
	A. niger	X	X	X	X	X	X	X	X
4th inoculation	3 types of bacteria	X	X	X	X	X	X	X	X
	C. albicans	X	X	X	X	X	X	X	X
	A. niger	X	X	X	X	X	X	X	X
X: No antibacterial activity (bacterial growth), O: Antibacterial activity (no bacterial growth)

Referring to Table 10, regardless of the formulation, it was confirmed that the cosmetic composition containing the preservative compound and luterin stably maintained antibacterial activity against bacteria and fungi without changing the formulation.

So far, the present invention has been looked at with respect to its preferred embodiments. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative rather than a limiting point of view. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent scope will be construed as being included in the present invention.

Claims (8)

1. An antibacterial composition comprising an antiseptic compound and luterin as active ingredients.
2. The method of claim 1,

   The antiseptic compound is an antimicrobial composition of at least one selected from the group consisting of alkane diol, hydroxyacetophenone, glyceryl caprylate, ethylhexyl glycerin and sodium benzoate.
3. The method of claim 2,

   The alkane diol is an antimicrobial composition of the alkane diol having 3 to 12 carbon atoms.
4. The method of claim 1,

   The antimicrobial composition is an antimicrobial composition having antibacterial activity against bacteria and fungi.
5. The method of claim 1,

   The antimicrobial composition is an antimicrobial composition comprising 0.01 to 50% by weight of the preservative compound based on the total weight.
6. The method of claim 1,

   The antimicrobial composition is an antimicrobial composition comprising 0.05 to 50 mM of luterin.
7. A cosmetic composition comprising an antiseptic compound and luterin as active ingredients.
8. A preservative for preserving cosmetics containing a preservative compound and luterin as active ingredients.

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