WO2022131613A1 - Cosmetic composition containing mixture of rosmarinic acid and lemon balm leaf extract as active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition which contains a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient, increases the expression of sodium hydrogen exchanger1 (NHE1), which is an internal factor that regulates skin pH, thus having the effect of maintaining weakly acidic skin and strengthening the skin barrier, and has various other beneficial effects on the skin.

Description

Cosmetic composition containing a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient

The present invention relates to a cosmetic composition, and more particularly, by containing a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient, by increasing the expression of NHE1 (Sodium hydrogen exchanger1), an internal factor that regulates skin pH, maintenance of weakly acidic skin and skin It relates to a cosmetic composition containing, as an active ingredient, a mixture of rosmarinic acid and lemon balm leaf extract, which has a barrier strengthening effect and has various other beneficial effects on the skin.

In general, the most important role of the skin is a function as a defense barrier that protects the body from external damage and invasion of bacteria. The skin is composed of three layers from the outside: the epidermis, the dermis, and the subcutaneous fat. Thus, it functions as the most important barrier to protect the skin from the external environment. Keratinocytes generated in the basal layer of the epidermis reach the stratum corneum and release abundant lipids from the lamellar body. Afterwards, the nucleus is lost and remains as a protein mass, corneocytes. . The stratum corneum is composed of keratinocytes (bricks) and the intercellular lipid membrane (mortar) that fills them, and performs a strong defense function to protect the human body from external physical, chemical, and biological stimuli.

The skin surface of a young and healthy person has a slightly acidic pH of about 4.5 to 5.5, and this weakly acidic pH contributes to maintaining the homeostasis of the skin barrier. However, the skin of the elderly has a slightly higher pH, so it becomes neutral or slightly alkaline. In such a weakly alkaline environment, the activity of enzymes involved in the formation of a lipid film is reduced, the corneodesmosome, which tightly binds keratinocytes, is decomposed, and the activity of serine proteases, an enzyme involved in exfoliation of dead skin cells, increases. Barrier dysfunction is caused. In aging skin, the expression of sodium-hydrogen exchanger-1 (NHE-1), which plays a decisive role in maintaining the acidic pH of the stratum corneum, is reduced. It is thought that the rate of recovery to normal after the skin barrier is damaged as a result of not maintaining the acidic pH is greatly reduced. Therefore, there has been a growing need for a cosmetic composition capable of adjusting skin pH.

As a prior art related to such pH control, Korean Patent No. 10-2014395 discloses a cosmetic composition comprising black cumin extract as an active ingredient, which is a skin pH control factor, sodium/hydrogen ion channel ( By increasing the activity of Na+/H+ Exchanger 1), black cumin (Nigella sativa), which is more essential and continuously maintains the skin pH in a weakly acidic state, improves the skin barrier function, and has an excellent effect on skin flexibility and moisturizing It relates to a cosmetic composition for adjusting skin pH comprising an extract as an active ingredient.

However, the conventional cosmetic composition containing black cumin extract as an active ingredient is difficult to obtain black cumin as a raw material, so there is a need for cosmetics for adjusting skin pH using raw materials that are easy to grow around us. In addition, the cosmetic composition comprising the conventional black cumin extract as an active ingredient had a slight pH control effect.

Therefore, it is easily available around us, but by increasing the expression of NHE1 (Sodium hydrogen exchanger1), an internal factor that regulates skin pH, it maintains weakly acidic skin and strengthens the skin barrier. The need for cosmetic compositions was growing.

[Prior art literature]

[Patent Literature]

(Patent Document 0001) Republic of Korea Patent Publication No. 10-2014395 (published on Aug. 26, 2019), "Cosmeceutical composition containing black cumin extract as an active ingredient"

The present invention relates to a cosmetic composition, and more particularly, by containing a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient, by increasing the expression of NHE1 (Sodium hydrogen exchanger1), an internal factor that regulates skin pH, maintaining weakly acidic skin and It relates to a cosmetic composition containing, as an active ingredient, a mixture of rosmarinic acid and lemon balm leaf extract, which has a skin barrier strengthening effect and has various other beneficial effects on the skin.

In order to achieve the above object, the present invention contains a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient to increase the expression of NHE1 (sodium hydrogen exchanger1), an internal factor that controls skin pH, rosmarinic acid and It provides a cosmetic composition containing a mixture of lemon balm leaf extract as an active ingredient.

In addition, the mixture of rosmarinic acid and lemon balm leaf extract is dissolved in powdered rosmarinic acid and lemon balm leaf extract in one or more solvents selected from the group consisting of purified water, ethanol, methanol, butanol, propanol, acetone, butylene glycol and hexanediol and stirred and mixed.

In addition, the rosmarinic acid and lemon balm leaf extract are mixed in a weight ratio of 0.01 to 10: 0.1 to 90.

In addition, the lemon balm leaf extract is characterized by hot water extraction with at least one extraction solvent selected from the group consisting of purified water, ethanol, methanol, butanol, propanol, acetone, butylene glycol and hexanediol.

In addition, it is characterized in that it contains a mixture of rosmarinic acid and lemon balm leaf extract in an amount of 0.001 to 90.0% by weight based on the total weight.

The cosmetic composition containing a mixture of rosmarinic acid and lemon balm leaf extract according to the present invention as an active ingredient contains a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient, and the expression of NHE1 (Sodium hydrogen exchanger1), an internal factor that controls skin pH It has the effect of maintaining weakly acidic skin and strengthening the skin barrier by increasing

1 is a graph showing the increase rate of NHE1 mRNA expression in Examples 1 to 3 and Comparative Examples 1 to 2;

2 is a graph showing the NHE1 protein production promotion rate of Examples 1 to 3 and Comparative Examples 1 to 2;

3 is a graph showing changes in skin pH over time after applying Preparation Example 2 and Comparative Preparation Examples 1 to 3;

4 is a graph showing the change in the amount of transdermal moisture loss over time after applying Preparation Example 2 and Comparative Preparation Examples 1 to 3;

5 is a graph showing the cell proliferation rate for the contaminants of Examples 1 to 3 and Comparative Examples 1 and 2;

6 is a graph showing the blackhead removal effect of Preparation Example 2 and Comparative Preparation Examples 1 to 3;

7 is a graph showing the collagen synthesis rate of Examples 1 to 3 and Comparative Examples 1 and 2;

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The following detailed description of the present invention is an embodiment in which the present invention may be practiced, and reference is made to the accompanying drawings shown by way of example of the embodiment. These embodiments are described in sufficient detail to enable those skilled in the art to practice the present invention. It should be understood that various embodiments of the present invention are different but need not be mutually exclusive. For example, specific shapes, structures, and characteristics described herein may be implemented in other embodiments without departing from the spirit and scope of the invention in relation to one embodiment. In addition, it should be understood that the location or arrangement of individual components within each described embodiment may be changed without departing from the spirit and scope of the present invention.

Accordingly, the detailed description set forth below is not intended to be taken in a limiting sense, and the scope of the present invention, if properly described, is limited only by the appended claims, along with all scopes equivalent to those claimed by the claims. Like reference numerals in the drawings refer to the same or similar functions throughout the various aspects.

The terms used in the present invention have been selected as currently widely used general terms as possible while considering the functions in the present invention, but these may vary depending on the intention or precedent of a person skilled in the art, the emergence of new technology, and the like. In addition, in a specific case, there is a term arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the corresponding invention. Therefore, the term used in the present invention should be defined based on the meaning of the term and the overall content of the present invention, rather than the name of a simple term.

In the present invention, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.

Hereinafter, a cosmetic composition containing a mixture of rosmarinic acid and lemon balm leaf extract according to the present invention as an active ingredient will be described in detail.

The cosmetic composition containing a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient according to the present invention contains a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient to control the skin pH. Expression of NHE1 (Sodium hydrogen exchanger1) It has various beneficial effects on the skin such as anti-pollution effect, blackhead removal effect and wrinkle improvement effect.

The rosmarinic acid is a kind of polyphenol compound abundantly present in various plants such as rosemary, sesame leaf, and lemon. As a polyphenol component, rosmarinic acid has been studied to have various pharmacological effects such as antioxidant effect. The rosmarinic acid has the form of Formula 1 below.

The rosmarinic acid is a caffeic acid ester of dihydroxyphenyl lactic acid [3- (3,4-dihydroxyphenyl) lactic acid]. The rosmarinic acid was first extracted and analyzed from rosemary by M. L. Scarpatti and G. Oriente in 1958. It is appropriate to use the rosmarinic acid in powder form, and various products are commercially available for the rosmarinic acid in powder form.

The lemon balm leaf extract refers to an extract extracted from the leaves of lemon balm ( Melissa officinalis ). The lemon balm is a cold-resistant perennial plant of the dicotyledonous plant Lamiaceae. The lemon balm is also called melisse citronnelle because it smells like lemon. The alcoholic extract of lemon balm is used as a raw material for l'eau des Carmes, a potion known to strengthen the heart and relieve headaches. The lemon balm can use all of flowers, leaves, and stems, and essential oil is used as an active substance.

On the other hand, the lemon balm leaf extract is preferably obtained by hot water extraction of lemon balm leaves into one or more extraction solvents selected from the group consisting of purified water, ethanol, methanol, butanol, propanol, acetone, butylene glycol and hexanediol. . More specifically, it may be obtained by putting leaves of lemon balm in an extraction solvent and heating at a temperature of 70 to 120° C. for 30 to 240 minutes.

The mixture of rosmarinic acid and lemon balm leaf extract refers to a mixture of powdered rosmarinic acid and lemon balm leaf extract dissolved in a solvent and stirred.

In this case, it is appropriate that the solvent is at least one selected from the group consisting of purified water, ethanol, methanol, butanol, propanol, acetone, butylene glycol and hexanediol. More suitably, it is purified water or butylene glycol.

On the other hand, the mixture of rosmarinic acid and lemon balm leaf extract is preferably dissolved in a solvent by mixing rosmarinic acid and lemon balm leaf extract in a weight ratio of 0.01:10 to 0.1:90. The remaining amount may include any one or more solvents selected from the group consisting of purified water, ethanol, methanol, butanol, propanol, acetone, butylene glycol and hexanediol.

On the other hand, the cosmetic composition containing the mixture of rosmarinic acid and lemon balm leaf extract according to the present invention as an active ingredient, in addition to the mixture of rosmarinic acid and lemon balm leaf extract, purified water, caprylic/capric triglyceride, cetearyl alcohol, dimethy Soybean, 1,2-exandiol, cetearyl olivate, sorbitan olivate, carbomer, tromethamine, ethylhexyl glycerin and citric acid may be formulated to further include. When formulating including the above ingredients, it will be preferable to include a mixture of rosmarinic acid and lemon balm leaf extract in an amount of 0.001 to 90.0% by weight based on the total weight of the composition, and other ingredients.

On the other hand, the cosmetic composition containing the mixture of rosmarinic acid and lemon balm leaf extract according to the present invention as an active ingredient increases the expression of NHE1 (Sodium hydrogen exchanger1), an internal factor that regulates skin pH, has a weakly acidic skin maintenance effect, It has a barrier strengthening effect.

In addition, the cosmetic composition comprising a mixture of rosmarinic acid and lemon balm leaf extract according to the present invention as an active ingredient contains a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient, thereby providing anti-pollution effect, blackhead removal effect and wrinkle improvement effect It has a variety of beneficial effects on the skin.

Hereinafter, the effect of the cosmetic composition containing the mixture of rosmarinic acid and lemon balm leaf extract according to the present invention as an active ingredient will be described in detail through the following Examples, Comparative Examples, Preparation Examples, Comparative Preparation Examples and Experimental Examples.

Examples 1 to 3. Preparation of a mixture of rosmarinic acid and lemon balm leaf extract

A mixture of rosmarinic acid and lemon balm leaf extract of Examples 1 to 3 was prepared through the following preparation steps.

(S10): Rosmarinic acid powder was prepared.

(S20): Lemon balm leaf extract was prepared by putting leaves of lemon balm in purified water and heating at a temperature of 90° C. for 120 minutes.

(S30): A mixture of rosmarinic acid and lemon balm leaf extract of Examples 1 to 3 was prepared by mixing 1,3-butylene glycol, rosmarinic acid powder, and lemon balm leaf extract in the composition ratio shown in Table 1 below and stirring.

	Mixed weight ratio (wt%)
	Example 1	Example 2	Example 3
1,3-butylene glycol	remaining amount	remaining amount	remaining amount
Lemon Balm Leaf Extract	50.0	50.0	50.0
Rosmarinic Acid Powder	0.1	1.0	2.0

Comparative Example 1. Preparation of a mixture containing only rosmarinic acid

A mixture containing only rosmarinic acid of Comparative Example 1 was prepared through the following preparation steps.

(S10): Rosmarinic acid powder was prepared.

(S20): A mixture containing only rosmarinic acid of Comparative Example 1 was prepared by mixing 1,3-butylene glycol with rosmarinic acid powder at the composition ratio shown in Table 2 below and stirring.

	Mixed weight ratio (wt%)
	Comparative Example 1
1,3-butylene glycol	remaining amount
Rosmarinic Acid Powder	1.0

Comparative Example 2. Preparation of a mixture containing only lemon balm leaf extract

A mixture containing only the lemon balm leaf extract of Comparative Example 2 was prepared through the following preparation steps.

(S10): Lemon balm leaf extract was prepared by putting leaves of lemon balm in purified water and heating at a temperature of 90° C. for 120 minutes.

(S20): A mixture containing only the lemon balm leaf extract of Comparative Example 2 was prepared by mixing the lemon balm leaf extract with 1,3-butylene glycol at the composition ratio shown in Table 3 below and stirring.

	Mixed weight ratio (wt%)
	Comparative Example 2
1,3-butylene glycol	remaining amount
Lemon Balm Leaf Extract	50.0

production example 1 to 3 and Comparative Preparation Example 1 to 3. Example 1 to 3 and comparative example Preparation of cosmetic formulations using 1-2

The mixtures prepared in Examples 1 to 3 and Comparative Examples 1 to 2 were prepared as cosmetic formulations.

Purified water, caprylic/capric triglyceride, cetearyl alcohol, dimethicone, 1,2-hexanediol, cetearyl olivate, and sorbate in the mixtures prepared in Examples 1 to 3 and Comparative Examples 1 to 2 Bitanolivate, carbomer, tromethamine, ethylhexyl glycerin and citric acid were further included and formulated as the cosmetics of Preparation Examples 1 to 3 and Comparative Preparation Examples 1 and 2 as shown in Table 4 below. On the other hand, Comparative Preparation Example 3 in which none of the mixtures prepared in Examples 1 to 3 and Comparative Examples 1 to 2 were included were also prepared.

	Mixed weight ratio (wt%)
	Preparation Example 1	Preparation Example 2	Preparation 3	Comparative Preparation Example 1	Comparative Preparation Example 2	Comparative Preparation Example 3
Purified water	remaining amount	remaining amount	remaining amount	remaining amount	remaining amount	remaining amount
Example 1	2.0	-	-	-	-	-
Example 2	-	2.0	-	-	-	-
Example 3	-	-	2.0	-	-	-
Comparative Example 1	-	-	-	2.0	-	-
Comparative Example 2	-	-	-	-	2.0	-
Caprylic/Caprylic Triglyceride	5.0	5.0	5.0	5.0	5.0	5.0
cetearyl alcohol	2.5	2.5	2.5	2.5	2.5	2.5
dimethicone	2.0	2.0	2.0	2.0	2.0	2.0
1,2-Hexanediol	1.35	1.35	1.35	1.35	1.35	1.35
cetearyl olivate	1.2	1.2	1.2	1.2	1.2	1.2
Sorbitan Oliveate	0.8	0.8	0.8	0.8	0.8	0.8
Carbomer	0.3	0.3	0.3	0.3	0.3	0.3
tromethamine	0.25	0.25	0.25	0.25	0.25	0.25
Ethylhexylglycerin	0.15	0.15	0.15	0.15	0.15	0.15
Citric Acid	0.02	0.02	0.02	0.02	0.02	0.02

Experimental Example 1. Confirmation of increase rate of NHE1 mRNA expression

For the mixtures prepared in Examples 1 to 3 and Comparative Examples 1 to 2, the increase rate of NHE1 mRNA expression was confirmed.

Normal human epidermal keratinocytes (NHEKs) are cultured in a 75 cm ² flask of Keratinocyte Growth Medium (KGM). Collect the cells by trypsinization and dilute with KGM to a concentration of 1.5 x 10 ⁵ cells/mL, then inoculate 2 mL of the cell mixture into a 6-well plate and incubate overnight. After that, KGM is exchanged with Keratinocyte Basal Medium (KBM) and the cells are cultured for 24 hours. Test samples of Examples 1 to 3 and Comparative Examples 1 and 2 dissolved in KBM by 1000-fold dilution are added to the wells, and after 24 hours of incubation, total RNAs are isolated according to a standard method. Total RNAs of cells cultured without the addition of Examples 1 to 3 and Comparative Examples 1 and 2 were isolated in the same manner. cDNA was synthesized by reverse transcription of 200ng total RNA, and then real-time PCR analysis was performed using Thermal Cycler Dice _® Real Time SystemⅢ (Takara Bio Inc, Otsu, Japan), TaKaRa SYBR _® PrimeScriptTM RT-PCR Kit (Perfect Real Time) (Takara Bio Inc, Otsu, Japan) and NHE1 or GAPDH (glyceraldehyde-3-phosphate) primer set. Relative expression is given as the ratio of expression of the NHE1 gene to the GAPDH gene (NHE1/GAPDH ratio). The enhancing effect on NHE1 mRNA expression was evaluated using the following formula.

NHE1 mRNA expression increase rate (%) = A / B x 100

A: NHE1/GAPDH ratio of test sample

B: NHE1/GAPDH ratio of control group

The results are shown in Table 5 below.

	NHE1 mRNA expression increase rate (%)
control	100%
Example 1	137%
Example 2	166%
Example 3	178%
Comparative Example 1	136%
Comparative Example 2	127%

As a result, it was confirmed that the NHE1 mRNA expression increase rate when rosmarinic acid and lemon balm leaf extract were mixed was significantly higher compared to Comparative Example 1 containing only rosmarinic acid and Comparative Example 2 containing only lemon balm leaf extract. It was confirmed that the efficacy was excellent when rosmarinic acid and lemon balm leaf extract were mixed, compared to the case of using the rosmarinic acid and the case of using only the lemon balm leaf extract alone.

Experimental Example 2. Confirmation of NHE1 protein production promotion rate

For the mixtures prepared in Examples 1 to 3 and Comparative Examples 1 to 2, the NHE1 protein production promotion rate was confirmed.

Normal human epidermal keratinocytes (NHEKs) are cultured in a 75 cm ² flask of Keratinocyte Growth Medium (KGM). Collect cells by trypsinization and dilute with KGM to a concentration of 1.5 x 10 ⁵ cells/mL, then inoculate 2 mL of the cell mixture into 6-well plates and incubate overnight. After that, KGM is exchanged with Keratinocyte Basal Medium (KBM) and the cells are cultured for 24 hours. Examples 1 to 4 and Comparative Examples 1 to 4 dissolved in KBM by 1000-fold dilution were added to the wells, and after 48 hours of incubation, the cells were incubated with 400 μL of 50 mmol/LTris-HCl buffer (pH7.5, 150 mmol/L NaCl, Dissolve in 1% NonidetP40, 0.5% sodium deoxycholate and protease inhibitor) and centrifuge to remove cell debris. The amount of NHE1 contained in 100 μL of the protein solution was measured with the NHE1 ELISA Kit (Cloud-Clone Corp.). Simultaneously, the amount of total protein was quantified in 20 μL of protein solution and the amount of NHE1 per total protein was calculated. The amount of NHE1 per total protein extracted from cells without the addition of Examples 1 to 4 and Comparative Examples 1 to 4 was set to 100, and the NHE1 production promoting effect per total protein extracted from cells of the test sample was evaluated using the following formula.

NHE1 protein production promotion rate (%) = A / B x 100

A: Amount of NHE1 per total protein extracted from cells containing the test sample

B: Amount of NHE1 per total protein extracted from cells without test sample

The results are shown in Table 6 below.

	NHE1 protein production promotion rate (%)
control	100%
Example 1	140%
Example 2	170%
Example 3	182%
Comparative Example 1	139%
Comparative Example 2	119%

As a result, it was confirmed that the NHE1 protein production promotion rate was significantly higher when rosmarinic acid and lemon balm leaf extract were mixed compared to Comparative Example 1 containing only rosmarinic acid and Comparative Example 2 containing only lemon balm leaf extract. It was confirmed that the efficacy was excellent when rosmarinic acid and lemon balm leaf extract were mixed compared to the case of using alone and the case of using only lemon balm leaf extract.

Experimental Example 3. Confirmation of skin pH and skin barrier improvement effect

For the cosmetic formulations prepared in Preparation Example 2 and Comparative Preparation Examples 1 to 3, the skin pH and skin barrier improvement efficacy was confirmed.

After 20 subjects applied the cosmetic formulations prepared in Preparation Example 2 and Comparative Preparation Examples 1 to 3 to the forearms of both arms twice a day, twice a day, IQ of 1.2% Sodium Lauryl Sulfate (SLS) solution was applied to the forearms. It was dropped into the chamber and applied to the test site for 24 hours. After artificially increasing skin pH and TEWL, the pH and transdermal water loss of the test site were measured on the 3rd and 1st weeks using a pH meter and a vapometer, respectively.

As a result, the skin pH is shown in Table 7 below, and the amount of transdermal water loss is shown in Table 8 below.

	skin pH
	before damage	immediately after damage		Day 3	Week 1
Preparation Example 2	4.98	5.47	5.21	5.06
Comparative Preparation Example 1	4.97	5.42	5.34	5.27
Comparative Preparation Example 2	4.96	5.45	5.32	5.25
Comparative Preparation Example 3	4.95	5.47	5.43	5.38

	transdermal water loss
	before damage	immediately after damage		Day 3	Week 1
Preparation Example 2	7.60	23.56	16.78	8.27
Comparative Preparation Example 1	7.62	23.54	18.54	9.58
Comparative Preparation Example 2	7.65	23.52	18.78	9.68
Comparative Preparation Example 3	7.64	23.51	19.60	9.89

Compared to Comparative Preparation Example 1 containing only rosmarinic acid, Comparative Preparation Example 2 containing only lemon balm leaf extract, and Comparative Preparation Example 3 containing nothing, when rosmarinic acid and lemon balm leaf extract were mixed, the pH and transdermal moisture increased by external stimulation It was confirmed that the loss was further reduced. Accordingly, it was confirmed that the efficacy was excellent when rosmarinic acid and lemon balm leaf extract were mixed, compared to the case where only rosmarinic acid was used alone and when only lemon balm leaf extract was used.

Experimental Example 4. Confirmation of anti-pollution effect

The anti-pollution effect was confirmed for the cosmetic formulations prepared in Examples 1 to 3 and Comparative Examples 1 to 2.

In general, the degree of cellular damage caused by pollutants was measured using skin keratinocytes that come in direct contact with pollutants, and Examples 1 to 3 and Comparative Examples 1 to 2 were treated, and the effect of inhibiting skin cell damage was confirmed. . The experiment was carried out according to a known method.

200 μl of human keratinocyte HaCaT at a concentration of 1×10 ⁴ cells/ml was dispensed into a 96-well plate and cultured at 37° C., CO ₂ in an incubator for 24 hours. In this case, IMDM medium containing 10% bovine serum and 1% antibiotic was used as the medium. After culturing, the medium was replaced with an IMDM medium excluding bovine serum and antibiotics, and then treated with benzopyrene (2.5 μg/mL), ammonium nitrate (500 ppm), and formaldehyde (1.5 μg/mL) in Examples 1 to 3 and Comparative Examples 1 to 2 were diluted 1000 times and further cultured for 21 hours. Thereafter, MTT solution of 5 mg/ml concentration was added and cultured for 3 hours. Then, the medium was removed and 150 μl of dimethyl sulfoxide was added to each well, followed by shaking for 10 minutes, and absorbance was measured at 570 nm. Cell viability of the experimental group not treated with contaminants was used as a control group, and the cell proliferation rate of the control group was 100%, and the cell proliferation rates of Examples 1 to 3 and Comparative Examples 1 and 2 were compared. In the comparison group, only purified water was added instead of the experimental group.

	Cell proliferation rate (%) for benzopyrene (2.5 μg/mL)	Cell proliferation rate (%) for ammonium nitrate (500 ppm)	Cell proliferation rate (%) for formaldehyde (1.5 μg/mL)
Control group (untreated contaminant group)	100	100	100
Comparative group (experimental group, untreated group)	42	43	41
Example 1	69	72	76
Example 2	87	89	82
Example 3	85	86	79
Comparative Example 1	66	67	65
Comparative Example 2	68	66	68

As a result, it was confirmed that the cell proliferation rate against contaminants was better when rosmarinic acid and lemon balm leaf extract were mixed compared to Comparative Example 1 containing only rosmarinic acid and Comparative Example 2 containing only lemon balm leaf extract. Accordingly, Compared to the case where only rosmarinic acid was used and the case where only the lemon balm leaf extract was used alone, it was confirmed that the efficacy was excellent when the rosmarinic acid and the lemon balm leaf extract were mixed when the rosmarinic acid and the lemon balm leaf extract were mixed.

Experimental Example 5. Confirmation of blackhead removal effect

The blackhead removal effect was confirmed for the cosmetic formulations of Preparation Example 2 and Comparative Preparation Examples 1 to 3.

The test was conducted according to a known method. The blackhead removal effect was evaluated when the cosmetic formulations prepared in Preparation Example 2 and Comparative Preparation Examples 1 to 3 were used for 20 subjects suffering from blackhead problems. 20 subjects were divided into each group and the cosmetic formulations of Preparation Example 2 and Comparative Preparation Examples 1 to 3 were applied for 2 weeks.

After 4 weeks, a makeup expert compared the skin condition images of the subjects before the experiment with the skin condition images of the subjects 4 weeks after the experiment, and evaluated the degree of blackhead removal using a five-point scale method. 10 is shown.

	Degree of blackhead removal (average)
Preparation Example 2	4.8
Comparative Preparation Example 1	3.6
Comparative Preparation Example 2	3.2
Comparative Preparation Example 3	2.4

As a result, compared to Comparative Preparation Example 1 containing only rosmarinic acid, Comparative Preparation Example 2 containing only lemon balm leaf extract, and Comparative Preparation Example 3 containing nothing, when rosmarinic acid and lemon balm leaf extract were mixed, the blackhead removal effect was better Accordingly, it was confirmed that when rosmarinic acid and lemon balm leaf extract were mixed, the efficacy was excellent when rosmarinic acid and lemon balm leaf extract were mixed, compared to when only rosmarinic acid was used alone and when only lemon balm leaf extract was used alone did

Experimental Example 6. Measurement of collagen synthesis rate

For Examples 1 to 3 and Comparative Examples 1 to 2, the collagen synthesis effect was confirmed.

The experiment was performed according to a known method. Examples 1 to 3 and Comparative Examples 1 to 2 were treated in human dermal fibroblasts to measure the collagen synthesis effect.

Human dermal fibroblasts were inoculated into each well of a 96-well microplate so as to become 1×10 ⁴ cells, and cultured in DMEM for 24 hours.

Human dermal fibroblasts (human dermal fibroblasts) were inoculated into a 96-well plate containing serum-containing DMEM medium (3 X 10 ³ cells/well) and incubated for 24 hours at 37° C. in a 5% concentration CO ₂ incubator. After removing the medium from each well, Examples 1 to 3 and Comparative Examples 1 to 2 were diluted 1000-fold, treated with a serum-free medium, and cultured again for 48 hours After 48 hours, the cell culture medium was collected and samples were collected. For the amount of collagen in the sample, the amount of Procollagen Type I C-peptide (PICP) was measured using a collagen measurement kit (Takara, Japan). The experiment was performed according to the method described in the manual. carried out.

The control group is that Examples 1 to 3 and Comparative Examples 1 and 2 are not treated, and the comparative group is Examples 1 to 3 and Comparative Examples 1 and 2 are not treated, and ultraviolet rays are irradiated. The results are as shown in Table 11 below, and the control group was 100%, and the remaining experimental groups were compared thereto.

	Collagen synthesis rate (%)
control	100
control group	76
Example 1	135
Example 2	175
Example 3	184
Comparative Example 1	130
Comparative Example 2	129

As a result, it was confirmed that the collagen synthesis rate was better when rosmarinic acid and lemon balm leaf extract were mixed compared to Comparative Example 1 containing only rosmarinic acid and Comparative Example 2 containing only lemon balm leaf extract. Through the collagen synthesis rate, the present invention It could be inferred that the cosmetic composition containing a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient had an improved anti-wrinkle effect.

Accordingly, it was confirmed that the efficacy was excellent when rosmarinic acid and lemon balm leaf extract were mixed when rosmarinic acid and lemon balm leaf extract were mixed, compared to when only rosmarinic acid was used alone and when only lemon balm leaf extract was used alone.

conclusion.

As confirmed through the Examples, Comparative Examples, Preparation Examples, Comparative Preparation Examples and Experimental Examples, the cosmetic composition containing the mixture of rosmarinic acid and lemon balm leaf extract according to the present invention as an active ingredient increases NHE1 mRNA expression and increases NHE1 protein It was confirmed that it promotes the production.

Accordingly, the cosmetic composition containing the mixture of rosmarinic acid and lemon balm leaf extract according to the present invention as an active ingredient increases the expression of NHE1 (Sodium hydrogen exchanger1), thereby maintaining a weakly acidic skin effect, preventing transdermal moisture loss, and strengthening the skin barrier effect and the like.

In addition, it was confirmed that the cosmetic composition containing the mixture of rosmarinic acid and lemon balm leaf extract according to the present invention as an active ingredient additionally has anti-pollution effect, blackhead removal effect and wrinkle improvement effect.

However, it was confirmed that rosmarinic acid and lemon balm leaf extract should be mixed in a weight ratio of 0.01: 90 to 0.1: 90 in order to express the above effects at a satisfactory level.

Therefore, the present invention has the effect of maintaining weakly acidic skin and strengthening the skin barrier by increasing the expression of NHE1 (Sodium hydrogen exchanger1), an internal factor that regulates skin pH, in addition to the anti-pollution effect, the blackhead removal effect and the wrinkle improvement effect It specifies that a cosmetic composition containing a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient has been developed as a novel composition with

Although the present invention has been described in conjunction with the accompanying drawings, this is only one embodiment of various embodiments including the gist of the present invention, and is intended to be easily implemented by those of ordinary skill in the art. For the purpose, it is clear that the present invention is not limited to the embodiments described above. Accordingly, the protection scope of the present invention should be interpreted by the following claims, and all technical ideas within the scope equivalent to that by changes, substitutions, substitutions, etc. within the scope not departing from the gist of the present invention are in the right of the present invention. will be included In addition, it is clarified that some components of the drawings are provided in an exaggerated or reduced form than in reality for more clearly explaining the configuration.

Claims (5)

1. A mixture of rosmarinic acid and lemon balm leaf extract, which contains a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient, is effective in controlling skin pH by increasing the expression of NHE1 (Sodium hydrogen exchanger1), an internal factor that controls skin pH. A cosmetic composition containing as an ingredient.
2. According to claim 1,

   The mixture of rosmarinic acid and lemon balm leaf extract is dissolved in any one or more solvents selected from the group consisting of purified water, ethanol, methanol, butanol, propanol, acetone, butylene glycol and hexanediol, and stirred A cosmetic composition containing a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient, characterized in that the mixture is mixed.
3. The method of claim 1,

   The cosmetic composition comprising a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient, characterized in that the rosmarinic acid and lemon balm leaf extract are mixed in a weight ratio of 0.01 to 10: 0.01 to 90.
4. The method of claim 1,

   The lemon balm leaf extract is a mixture of rosmarinic acid and lemon balm leaf extract, characterized in that it is hot-water extracted with at least one extraction solvent selected from the group consisting of purified water, ethanol, methanol, butanol, propanol, acetone, butylene glycol and hexanediol A cosmetic composition containing as an active ingredient.
5. The method of claim 1,

   A cosmetic composition comprising a mixture of rosmarinic acid and lemon balm leaf extract as an active ingredient, characterized in that it contains 0.001 to 90.0% by weight of a mixture of rosmarinic acid and lemon balm leaf extract based on the total weight.

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