WO2013077574A1

WO2013077574A1 - Topical composition containing theanine derivatives for use on skin

Info

Publication number: WO2013077574A1
Application number: PCT/KR2012/009377
Authority: WO
Inventors: 이진영; 유재원; 최향태; 김한별; 이성훈; 김지성; 배지현
Original assignee: 주식회사 아모레퍼시픽
Priority date: 2011-11-23
Filing date: 2012-11-08
Publication date: 2013-05-30

Abstract

The present invention relates to a topical composition for use on skin containing theanine derivatives as active components and, more particularly, to a topical composition for use on skin having anti-aging, skin wrinkle repair, skin barrier enhancement, skin immunity enhancement, skin moisturizing, and atopy alleviation effects by containing theanine derivatives as active components.

Description

External skin composition containing deanine derivative

The present invention relates to an external composition for skin containing a deanine derivative as an active ingredient, and more specifically, to an anti-aging, skin wrinkle improvement, skin barrier enhancement, skin immunity, skin moisturizing and The present invention relates to a topical skin composition having an atopic improvement effect.

Theanine is an amino acid contained in green tea (Camellia sinesis), which is produced by the enzymatic reaction of glutamine (L-glutamine) and ethylamine (ethylamine). Occupy and are mainly present in the form of free amino acids. Deanine inhibits caffeine-induced excitability (Toxicol Lett. 2001 123 (2-3), 159-167, Biosci. Biotechnol. Biochem. 2000 Feb; 64 (2): 287-93). alpha-waves) to relieve tension (Nippon Nogeikagaku Kaishi 1998; 72: 153-157) and antistress actions such as suppressing stress-induced heart rate increases and reducing stress response factors (Biol Psychol. 2007 74 (1): 39-45). It also lowers glutathione levels in cancer cells (Cancer Lett. 2004 212 (2), 177-184), lowers blood pressure (Biosci Biotechnol Biochem 1995 59 (4) 615-618), and low density lipoprotein cholesterol ( LDL-cholesterol) also has some antioxidant effects (Exp Toxicol Pathol 1997; 49: 329-335), and recently has anti-obesity effects (In Vivo 2004, 18 (1) 55-62) and alcohol degrading enzymes. Biological activities such as increase (Biol Pharm Bull 2005 28 (9) 1702, 1706) are also being studied.

The possibility of deanin having various physiological activities and specific studies have been made, but in the skin area, the lipolytic effect of deanine and green tea component catechins alone or in combination (Korean Patent Publication No. 2004-0092538) Except for) and skin aging improvement (Korean Patent Publication No. 2007-0028901), and the proline recycling promoting effect (Korean Patent Publication No. 2009-0064743), to date, known research cases and actions on the skin efficacy of deanine There is no report on the mechanism, and US patent application US2006 / 0134095 A1 describes the moisturizing effect of deanine and describes the deanine derivative as a cell activator, but the excellent skin efficacy of deanine derivative, not deanine itself. Is not known to date.

Accordingly, the present inventors have found that anti-aging, skin wrinkle improvement, skin barrier enhancement, skin immunity enhancement, skin moisturizing and atopy improvement as a superior and discriminating physiological effect beyond the imitation of the effect of deanine nucleus due to the physiological activity of the deanine derivative. It has been found to have an effect and the present invention has been completed.

Accordingly, an object of the present invention is to provide an external composition for skin having excellent anti-aging, skin wrinkle improvement, skin barrier enhancement, skin immunity, skin moisturizing and atopic improvement effect by containing a deanine derivative as an active ingredient.

The present invention relates to an anti-aging, skin wrinkle improvement, skin barrier reinforcement, skin immunity enhancement, skin moisturizing and atopy improvement composition containing a deanine derivative as an active ingredient.

The external preparation composition for skin containing the deanine derivative of the present invention is more excellent and differentiates the physiological effect of the deanine nucleus due to the physiological activity of the deanine derivative as anti-aging, skin wrinkle improvement, skin barrier strengthening, It has the effect of strengthening skin immunity, moisturizing skin and improving atopic dermatitis.

1 is a graph showing the expression rate of keratin 1 gene by deanine and the deanine derivative of the present invention.

Figure 2 is a graph showing the transglutaminase 1 gene expression rate by the deanine and the dean derivative of the present invention.

3 is a graph showing the expression rate of filaggrin gene by deanine and the deanine derivative of the present invention.

Figure 4 is a result showing the intracellular calcium migration pattern when treated with deanine and the dean derivative of the present invention.

Hereinafter, the present invention will be described in more detail.

The deanine derivative used in the present invention has a structure represented by the following general formula (1) or (2).

Formula 1

Wherein R ₁ is selected from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl and tert-butyl, and R2 is hydrogen, methyl, ethyl, propyl, isopropyl, n- Butyl, isobutyl and tert-butyl.)

Formula 2

The deanine derivative of the present invention is preferably contained in 0.0001 to 20% by weight based on the total weight of the composition. If it is less than 0.0001% by weight can not expect the desired effect, if it exceeds 20% by weight because it is not easy to control the viscosity of the cosmetic formulation.

The epidermal layer produces a complete stratum corneum through proliferation and differentiation and has homeostasis that drops off at the right time. In this process, keratinocytes that divide in the basal layer are transformed into keratinocytes by synthesizing various proteins and lipids to form a keratinous layer. Differentiation in the basal cells of the epidermis leads to the disappearance of the nucleus and intracellular organs, flattening and increasing the keratin intermediate fibers. In this differentiation, intracellular calcium concentration is important. In the lower epidermis with low calcium concentration, the proliferation of keratinocytes is promoted. In the upper epidermis with high calcium concentration, proliferation is suppressed and differentiation occurs. The keratin protein constituting the keratin intermediate microfibers is expressed in keratin5, keratin 14 in the basal layer and keratin 1, keratin 10 in the granule layer. Thus keratin 1 is used as an indicator for epidermal differentiation.

Aging of the skin is divided into photo-aging and natural aging, all of which are different from the epidermis of young skin. In photoaging, the thickness of the epidermis is abnormally thickened. In natural aging, the thickness of the epidermis decreases and the formed keratinocytes do not fall off well. All of this is due to lack of normal turn-over of the epidermis and inadequate differentiation control. These changes in epidermis are also involved in the formation of wrinkles in photoaging, according to a paper (JDS, 2001, Kambaytashi) that studies the mechanism of wrinkle formation by irradiating chronic low-dose UV radiation on hairless mice. According to the study, UV-irradiated skin produces wrinkles, and the amount of keratin 1 and filaggrin is significantly reduced.

The deanine derivative of the present invention has the effect of increasing the expression of keratin 1, this effect may result in the improvement of the homeostasis of the epidermal proliferation and differentiation, and through such homeostasis not only improve the natural aging skin but also ultraviolet light It can prevent or alleviate the formation of wrinkles.

One of the most important functions that skin performs is its function as a barrier against the free movement of various substances inside and outside the skin. As the skin barrier rapidly transitions from the granular layer to the stratum corneum, the keratin intermediate fine fibers are entangled to form a bundle of fibers, forming a stratum corneum that surrounds the inside of the cell membrane, cross-linking with protein components and other intracellular proteins in the stratum corneum The cell membrane is replaced. The keratin protein membrane and the keratin lipid membrane are combined to form a cornified envelope that forms the boundary of keratinocytes and completes the complete skin barrier structure. Filaggrin promotes the binding between keratin microfibers in the stratum corneum and granules to strengthen the barrier. Transglutaminase 1 crosslinks the various proteins necessary for the formation of the thick film. The long chain hydroxyceramide is linked to proteins to contribute to the formation of keratin lipid membranes.

The deanine derivative of the present invention is excellent in increasing the expression of Filaggrin (Filaggrin), Transglutaminase 1 (Transglutaminase 1). Therefore, the deanine derivative of the present invention has the effect of strengthening the barrier function, which is the main function of the skin, by helping to form the keratin thickening membrane, thereby strengthening the physical protective film of the skin, thereby enhancing the innate immunity of the skin.

Natural moisturizing factor (NMF) is one of the important factors for skin moisturizing and consists of amino acids and its derivatives. It contains moisture and keeps skin moisturized. In addition, the strengthening effect of the skin barrier is also a factor contributing to the skin moisturizing effect. Filaggrin not only functions to strengthen the barrier by promoting the binding between keratin microfibers in the stratum corneum and granules, but also breaks down into amino acids to form natural moisturizing factors. There are also periodic reports that the Filaggrin gene abnormality may be associated with the outbreak of atopic dermatitis. Thus, filaggrin not only enhances epidermal barrier function but also acts as a natural moisturizing factor, and is also involved in atopic dermatitis.

The deanine derivative of the present invention increases the expression of filaggrin as described above, and due to this effect, it can show skin moisturizing effect through increasing the natural moisturizing factor along with enhancing epidermal barrier function, and atopy due to the decrease of filaggrin. It has the effect of alleviating the symptoms of dermatitis.

The cosmetic composition is not particularly limited in formulation, and may be appropriately selected as desired. For example, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisturizing cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing It may be prepared in any one or more formulations selected from the group consisting of foam, cleansing lotion, cleansing cream, body lotion and body cleanser, but is not limited thereto.

When the formulation of the present invention is a paste, cream or gel, animal carriers, vegetable fibers, waxes, paraffins, starches, tracantes, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide, etc. may be used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, and especially in the case of spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspension agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

The cosmetic composition may further include functional additives and components included in the general cosmetic composition in addition to the added deanine derivative. For example, the functional additive may include a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extracts.

In addition to the said functional additive, you may mix | blend the component contained in a general cosmetic composition with the cosmetic composition of this invention as needed. In addition to the other components included, oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, blood circulation Accelerators, cooling agents, limiting agents, purified water and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples, but the scope of the present invention is not limited thereto.

Test Example 1 Confirmation of Keratin1 Gene Expression in Human Neonatal Epithelial Keratinocytes (HEK)

Human neonatal epithelial keratinocytes (HEK; Lonza, NHEK-Neo-Neonatal Normal Human Epidermal Keratinocytes, Pooled) were dispensed in 6-well medium (KBM-gold, Lonza) each at 1 × 10 ⁵ for 24 hours at 37 ° C. The culture was discarded, and the deanine derivatives were treated with a medium containing 100 μM and 500 μM (KBM-gold, Lonza, respectively) and cultured at 37 ° C. for 5 days.

RNA was extracted from the keratinocytes cultured for 5 days using RNeasy mini kit (qiagen), and cDNA was synthesized by RT-PCR with Superscript III kit (invitrogen). In order to confirm the expression level of Keratin 1, quantitative real-time PCR was performed using a probe (TaqManTM fluorogenic probe, Hs00196158_m1), and the expression patterns of Keratin 1 were observed, and the results are shown in FIG. 1. As a control, 100 μM and 500 μM deanine (Sigma, T6576) were used instead of the untreated group and the deanine derivative, respectively.

In the results of FIG. 1, it was confirmed that the deanine derivative increased the expression of Keratin 1 as compared to the untreated group, and the deanine derivative had a very excellent effect at the same concentration treatment as compared to the case where the deanine nucleus itself was treated. It can be seen that.

[Test Example 2] Transglutaminase 1 gene expression was confirmed in human neonatal epithelial keratinocytes (HEK)

RNA was extracted from the keratinocytes cultured for 5 days using RNeasy mini kit (qiagen), and cDNA was synthesized by RT-PCR with Superscript III kit (invitrogen). In order to confirm the expression level of transglutaminase 1, the expression pattern of transglutaminase was observed by performing quantitative real-time PCR using a probe (TaqManTM fluorogenic probe, Hs00165929_m1). Shown in As a control, 100 μM and 500 μM deanine (Sigma, T6576) were used instead of the untreated group and the deanine derivative, respectively.

From the results of FIG. 2, it was confirmed that the deanine derivatives increased the expression of transglutaminase 1 as compared to the untreated group, and the deanine derivatives were highly treated at the same concentration as compared to the case where the deanine nucleus itself was treated. It can be seen that the excellent effect.

Test Example 3 Confirmation of Filaggrin Gene Expression in Human Neonatal Epithelial Keratinocytes (HEK)

RNA was extracted from the keratinocytes cultured for 5 days using RNeasy mini kit (qiagen), and cDNA was synthesized by RT-PCR with Superscript III kit (invitrogen). In order to confirm the expression level of filaggrin, a quantitative real-time PCR was performed using a probe (TaqManTM fluorogenic probe, Hs00856927_g1), and the expression patterns of transglutaminase were observed. The results are shown in FIG. 3. As a control, 100 μM and 500 μM deanine (Sigma, T6576) were used instead of the untreated group and the deanine derivative, respectively.

In the results of FIG. 3, it was confirmed that the deanine derivative increased the expression of filaggrin compared to the untreated group, and the deanine derivative had a very excellent effect at the same concentration treatment as compared to the case of treating the deanine nucleus itself. It can be seen that.

Test Example 4 Evaluation of Calcium Influx in Human Neonatal Epithelial Keratinocytes (HEK)

Stabilized human neonatal epithelial keratinocytes (HEK; Lonza, NHEK-Neo-Neonatal Normal Human Epidermal Keratinocytes, Pooled) were obtained by 2x10 ⁴ in confocal dishes, 2.5 mM deanine and deanine derivative 0.5. After 5 minutes of mM treatment, 1 μm fluo 3-AM was treated for 15 minutes to detect intracellular calcium fluoride by confocal microscopy (Nikon, Japan). It evaluated, and the result is shown in FIG.

In the results of FIG. 4, it can be seen that the deanine and the deanine derivatives clearly show different results in intracellular calcium migration. These differences indicate that the deanine derivatives of the present invention do not merely mimic the effects of the deanine nucleus, but have new physiological activity effects different from that of the deanine nucleus.

Example 1 and Comparative Example 1

To prepare Example 1 and Comparative Example 1 according to the composition of Table 1.

Table 1

Compounding ingredients (unit: wt%)	Example 1	Comparative Example 1
Purified water	69	70
Propylene glycol	30	30
Deanine derivatives	One	-

Test Example 5 Measurement of Skin Barrier Function Restoration Effect

In order to measure the effect of the deanine derivative on the recovery of damaged skin barrier function due to skin damage, the following experiment was performed. Ten adult men and women's upper arms were damaged by the skin stripping method using a tape stripping method, and Comparative Example 1 and Example 1 were applied to a 3 cm × ³ cm area of 2 mg / cm ² daily for 7 days. The recovery of transdermal moisture loss (TWEL) was measured once a day using a Vapometer (Delfin, Finland). Percutaneous moisture loss was calculated using 100% transdermal moisture before tape stripping, and the results are shown in Table 2 below.

TABLE 2

Test group	TWEL change (%)
Test group	Before treatment	1 day	2 days	3 days	4 days	5 days	6 days
Example 1	100	86	73	54	31	31	31
Comparative Example 1	100	120	113	97	70	61	43

In the results of Table 2, when treating Example 1 containing a deanine derivative, it can be seen that the transdermal moisture loss returns to normal at a rapid rate and the barrier damage is recovered.

Example 2 and Comparative Example 2

To prepare a formulation of the nutrition cream of Example 2 and Comparative Example 2 according to the composition ratio of Table 3.

TABLE 3

Compounding Ingredients (wt%)	Example 2	Comparative Example 2
Purified water	To 100	To 100
Deanine derivatives	One	-
Vegetable Cured Oil	1.50	1.50
Stearic acid	0.60	0.60
Glycerol Stearate	1.00	1.00
Stearyl alcohol	2.00	2.00
Polyglyceryl-10 Pentastearate & Behenyl Alcohol & Sodium Stearoyl Lactylate	1.00	1.00
Arakidil Behenyl Alcohol & Arakidil Glucoside	1.00	1.00
Cetylaryl Alcohol & Cetearyl Glucoside	2.00	2.00
PEG-100 Stearate & Glycerolate & Propylene Glycol	1.50	1.50
Caprylic / Carric Triglycerides	11.00	11.00
Cyclomedicon	6.00	6.00
Preservative, incense	Quantity	Quantity
Triethanol amine	0.1	0.1

Test Example 6 Measurement of Effect of Increased Skin Moisturizing Power

In order to determine the effect of the deanine derivative on the skin moisturizing power, the following experiment was performed.

Sixty men and women in their 40s and 50s classified as dry skin were divided into two groups of 30 people each, and the nutrition creams of the two groups according to Example 2 and Comparative Example 2 were applied to the face twice daily for four weeks. Skin moisture meter at constant temperature and humidity conditions (24 ° C, 40% relative humidity) at the start of application, at 1 week, 2 weeks, 4 weeks after application, and 2 weeks after application termination (total 6 weeks). Skin moisture content was measured by (Corneometer CM825, C + K Electronic Co., Germany). The increase rate of the skin moisture content at each time point was calculated as a percentage based on the skin moisture content before the start of application, and the results are shown in Table 4 below.

Table 4

Test group	Moisture Growth Rate (%)
Test group	1 week past	2 weeks past	4 weeks past	After 6 weeks
Example 2	33	40	43	34
Comparative Example 2	30	32	31	22

In the results of Table 4, the application of Comparative Example 2 shows a water increase rate of about 30% up to 4 weeks after the application, but after stopping the application, the amount of skin moisture decreases, but it contains the deanine derivative. In the case of applying Example 2, it can be seen that the skin moisture increase rate is more than 30% even after the application is stopped. Through this it can be seen that the skin moisturizing effect of the composition according to the invention containing the deanine derivative is very excellent.

Test Example 7 Confirmation of Skin Wrinkle Improvement Efficacy

In order to confirm the skin wrinkle improvement effect of the composition according to the present invention was carried out the following experiment.

After dividing 80 healthy women in their 40s into two groups of 40 people, apply the nutrition cream according to the two groups of Example 2 and Comparative Example 2 once a day to the face for 12 weeks, and then use a silicone to float the replica The condition was measured by a skin meter (visiometer, SV600, Courage + Khazaka electronic GmbH, Germany) for image analysis. The results are shown in Table 5 below. The values in Table 5 below represent the average of each parameter value after 12 weeks of application minus the parameter value before application.

Table 5

Clinical results after 8 weeks of use	R1	R2	R3	R4	R5
Comparative Example 2	0.28	0.20	0.26	0.03	0.03
Example 2	-0.19	-0.10	-0.16	-0.03	-0.03
R1: The difference between the highest and lowest values of the wrinkled contours R2: The randomly divided wrinkles contours by 5 spaces Rn: Average of R1 values among them R3: The highest value of R1 values divided by 5 R4: Each at the baseline of the wrinkled contours R5: difference between the baseline of the wrinkle contour and the angle of each wrinkle

From the results of Table 5, it was found that the external preparation composition of Example 2 has a very good skin wrinkle improvement effect. Therefore, it can be confirmed that the cosmetic composition containing the deanine derivative of the present invention is very effective for improving skin wrinkles.

The anti-aging, skin wrinkle improvement, skin barrier strengthening, skin as a superior and discriminating physiological efficacy beyond the mimic the effect of the deanine nucleus due to the physiological activity of the deanine derivative by containing the deanine derivative of the present invention as an active ingredient The external preparation composition for skin which has an effect of strengthening immunity, moisturizing skin and improving atopic dermatitis can be provided.

Claims

A skin external composition comprising a deanine derivative having the structure of Formula 1 or Formula 2 as an active ingredient.

[Formula 1]

Wherein R 1 is selected from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl and tert-butyl, and R2 is hydrogen, methyl, ethyl, propyl, isopropyl, n- Butyl, isobutyl and tert-butyl.)

[Formula 2]

Wherein R 1 is selected from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl and tert-butyl, and R2 is hydrogen, methyl, ethyl, propyl, isopropyl, n- Butyl, isobutyl and tert-butyl.)

Partial external preparation composition.
The composition for external application for skin according to claim 1, wherein the deanine derivative is contained in an amount of 0.0001 to 20% by weight based on the total weight of the composition.
The composition for applying the external of the skin according to claim 1 or 2, wherein the composition promotes keratin 1 gene expression.
3. The composition for external application for skin according to claim 1 or 2, wherein the composition promotes the expression of Transglutaminase 1 gene.
3. The composition for external application for skin according to claim 1 or 2, wherein the composition promotes filaggrin gene expression.
The composition for external application for skin according to claim 1 or 2, wherein the composition regulates intracellular calcium migration.
The composition for external application for skin according to claim 1 or 2, wherein the composition is for anti-aging.
The composition for applying the external of the skin according to claim 1 or 2, wherein the composition is for improving skin wrinkles.
The composition for external application of skin according to claim 1 or 2, wherein the composition is for improving epidermal homeostasis maintenance.
The composition for external application for skin according to claim 1 or 2, wherein the composition is for strengthening a skin barrier.
The composition for external application for skin according to claim 1 or 2, wherein the composition is for strengthening skin immunity.
The composition for applying the external of the skin according to claim 1 or 2, wherein the composition is for moisturizing the skin.
The composition for applying the external of the skin according to claim 1 or 2, wherein the composition is for improving atopic dermatitis.