KR101373715B1 - Cosmetic composition promoting cornified envelope formation - Google Patents

Abstract

The present invention relates to a composition for promoting keratin film formation comprising citronellinic acid, derivatives thereof or salts thereof as an active ingredient. The present invention also relates to a composition for promoting keratin film formation comprising caffeic acid, its derivatives or salts thereof as an active ingredient.

Description

Cosmetic composition for promoting keratin film formation {COSMETIC COMPOSITION PROMOTING CORNIFIED ENVELOPE FORMATION}

The present invention relates to a cosmetic composition for promoting corneal film formation.

The skin is composed of the dermis and the epidermis, and the epidermis is located at the outermost part of the skin, and the stratum corneum composed of keratinocytes is normally formed and maintained, thus protecting against external physical, chemical and mechanical stimuli, and excessive body moisture through the skin. It protects against divergence. Keratinocytes formed through the morphological and functional changes in stages during the continuous transfer of basal cells from the stratum basale to the stratum corneum are removed from the skin after a certain period of time. New keratinocytes from the epidermal bottom layer repeat the process of epidermis differentiation or keratinization to take over their function. In this keratinization process, keratinocytes produce natural moisturizing factors and intercellular lipids such as ceramides, cholesterol and fatty acids, and the stratum corneum acts as a barrier to the outside, thus acting as a skin barrier. Will hold.

Dry skin, which is considered to be one of the major diseases in modern society, has been found to be the most important cause of abnormal skin barrier function. Such dry skin has recently increased due to various factors such as environmental pollution, increase of dry environment such as apartments and high-rise buildings, increase of social stress, excessive bathing culture unique to Korea, atopy and aging, etc. This is most often the case.

Therefore, a lot of researches have been conducted to supply moisture from the outside or to minimize moisture loss from the body in order to maintain proper skin moisture, and a moisturizing agent such as ceramide or a derivative thereof, which has a water retention ability, has been developed and the cosmetic field is developed. It is used a lot in. However, most of them do not show sufficient effect by temporary relief of symptoms rather than fundamental treatment. Therefore, it is urgent to develop a material that fundamentally regenerates a damaged barrier.

When PPAR (Peroxisome Proliferator Activated Receptor) present in keratinocytes is activated by binding to its ligand, it is known that it promotes differentiation of keratinocytes and reconstructs damaged skin barriers. PPAR alpha plays an important role in regulating cell proliferation / differentiation, regulation of inflammatory mediators, and has been reported to play an important role in regulating glucose, lipids, and hormone metabolism. In particular, it has been found to play an important role in maintaining homeostasis of skin function by promoting keratinocyte differentiation, inhibiting proliferation, promoting skin barrier formation through lipid metabolism and inhibiting inflammatory responses in the epidermis of the skin. In addition, the activity of PPAR alpha has been reported to play an important role in wound healing of skin. In fact, clofibrate, the known agonist of PPAR alpha [Feingold et al., J. Invest. Dermatol., 110, pp 368-375, 1998, WY14643 [Feingold et al., J. Invest. Dermatol., 115, pp353-360, 2000], when applied to the skin damaged skin barrier, it has been confirmed that the differentiation of keratinocytes is promoted, the recovery of the skin barrier is promoted.

The present inventors are studying the components that promote the activity of PPAR alpha to restore the skin barrier, while citronellinic acid and caffeic acid promote the activity of PPAR alpha to promote skin keratin formation to restore the skin barrier. After confirming that the present invention was completed.

An object of the present invention is to provide a composition for promoting keratin film formation.

In order to achieve the above object, the present invention provides a composition for promoting keratin film formation comprising citronellinic acid, its derivatives or salts thereof as an active ingredient.

The present invention also provides a composition for promoting keratin film formation comprising caffeic acid, its derivatives or salts thereof as an active ingredient.

The composition of the present invention has PPAR alpha activity promoting ability, epidermal cell differentiation promoting ability, thereby promoting the formation of keratinocytes to strengthen the skin barrier, moisturize the skin and relieve skin irritation.

1 shows the PPAR alpha activity enhancing ability of citronellic acid racemic mixture, caffeic acid, (S)-(-)-citronelic acid and (R)-(+)-citronellinic acid.
2 shows the concentration dependent PPAR alpha activity enhancing ability of the citronellinic acid racemic mixture.
Figure 3 shows the concentration-dependent PPAR alpha activity enhancing ability of caffeic acid.
Figure 4 shows the ability to promote the formation of keratin film of the citronellic acid racemic mixture, caffeic acid, (S)-(-)-citronellic acid and (R)-(+)-citronellinic acid.
5 shows the concentration dependent keratin formation promoting ability of the citronellinic acid racemic mixture.
Figure 6 shows the concentration-dependent keratin formation promoting ability of caffeic acid.
7 shows the ability to enhance involuclin expression of caffeic acid, and FIG. 8 shows the ability to enhance transglutaminase-1 expression of caffeic acid.
9 shows the filaggrin expression enhancing ability of caffeic acid.
10 shows the antioxidant capacity of caffeic acid.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention, and the manner of achieving them, will be apparent from and elucidated with reference to the embodiments described hereinafter in conjunction with the accompanying drawings. It should be understood, however, that the invention is not limited to the disclosed embodiments, but is capable of many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, To fully disclose the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.

The present invention relates to a composition for promoting keratin film formation comprising citronellinic acid, derivatives thereof or salts thereof as an active ingredient.

The present invention also relates to a composition for promoting keratin film formation comprising caffeic acid, its derivatives or salts thereof as an active ingredient.

Hereinafter, the present invention will be described in detail.

Citronellinic acid

Citronellic acid (Citronellic acid, 3,7-Dimethyloct-6-enoic acid) of the present invention is represented by the following formula (1). The citronellinic acids include their individual optical isomers, that is, (R)-(+)-citronellinic acid or S)-(-)-citronellinic acid, non-racemic mixtures thereof, or racemic mixtures. do. Formula 2 below represents (R)-(+)-citronelic acid, and Formula 3 below represents (S)-(-)-citronelic acid.

≪ Formula 1 >

(2)

(3)

Caffeic acid

Caffeic acid and derivatives thereof of the present invention are represented by the following formula (4). At this time, R1 or R2 is independently of each other hydrogen, a hydroxyl group or alkoxy having 5 or less carbon atoms. The alkoxy is preferably alkoxy having 3 or less carbon atoms, more preferably methoxy or ethoxy. R1 and R2 are preferably hydroxyl groups.

≪ Formula 4 >

Composition for promoting keratin film formation

The composition for promoting corneal film formation of the present invention has epidermal cell differentiation promoting ability, PPAR alpha activity promoting ability, skin stimulating alleviation ability, antioxidant activity, skin moisturizing ability or skin barrier strengthening ability. The composition of the present invention promotes the differentiation of epidermal cells to form keratinocytes, thereby promoting skin moisturization and preventing, improving and treating diseases caused by dry skin such as itching. The composition for promoting keratin film formation of the present invention is a cosmetic composition or a pharmaceutical composition.

In particular, the composition comprising the citronellinic acid, derivatives or salts thereof of the present invention as an active ingredient has the ability to alleviate skin irritation, promote epidermal cell differentiation, enhance the activity of PPAR alpha or enhance the skin barrier. In addition, the composition comprising the caffeic acid, derivatives or salts thereof of the present invention as an active ingredient, the ability to promote epidermal cell differentiation, PPAR alpha activity, antioxidant, skin irritation, skin moisturizing or skin barrier strengthening Has

Cosmetic composition

The cosmetic composition of the present invention may be prepared in any formulations conventionally produced in the art and may be formulated into a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant- containing cleansing oil, powder foundation , An emulsion foundation, a wax foundation and a spray, but is not limited thereto. The cosmetic composition of the present invention may also be formulated as a softening agent, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

Pharmaceutical composition

The pharmaceutical composition of the present invention has the effect of preventing and treating dry skin, itching, skin irritation and atopy. Therefore, the pharmaceutical composition of the present invention can be administered to patients with sensitive skin, dry skin, atopic skin, and the like, and can also be administered for the purpose of preventing such skin troubles.

The pharmaceutical composition of the present invention may comprise 0.01 to 80 parts by weight of citronellinic acid or caffeic acid, derivatives thereof or acceptable salts thereof, based on 100 parts by weight of the composition, preferably 0.02 to 65 parts by weight. It may include. However, this may be increased or decreased according to the needs of the administer, and may be appropriately increased or decreased depending on the condition of the skin and the progress of the itch.

The pharmaceutical composition of the present invention can be administered orally or parenterally, but is preferably applied to the skin and administered for transdermal absorption. The pharmaceutical composition of the present invention may be used in the form of a general pharmaceutical preparation or in the form of an emulsion for skin application. The pharmaceutical compositions of the present invention can be prepared using conventional carriers that are acceptable for pharmaceutical or cosmetic use. For example, in the case of preparations for oral administration, excipients, binders, disintegrants, glidants, solubilizers, suspending agents, preservatives or extenders may be used .

The dosage of the pharmaceutical composition comprising the citronellinic acid or caffeic acid, derivatives thereof or acceptable salts thereof of the present invention may be determined by a specialist depending on various factors such as the skin condition and age of the patient, but generally It may be administered at a dose of 0.01 g to 200 g, preferably 0.01 g to 50 g per cm ^{2 of} skin. It is also intended to contain a daily dose of 1/2 or 1/3 or 1/4 of the pharmaceutical composition, and may be administered 1 to 6 times a day. However, in the case of long-term administration for the purpose of maintaining skin health and hygiene, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

<Material>

In the following experimental example, a citronellic acid racemic mixture, a (S)-(-)-citronellic acid, a (R)-(+)-citronellinic acid treatment group, and a caffeic acid of the following Chemical Formula 5 were used.

&Lt; Formula 5 >

Experimental Example 1 Skin Barrier Strengthening Capability

<1-1> PPAR alpha activity enhancing ability

CV-1 cells, which are monkey kidney epithelial cell lines, were passaged in DMEM medium containing 10% fetal calf serum. Specifically, it includes a Renilla-Luciferase construct for control use to standardize the degree of transfection, and a peroxisome proliferator-responsive element (PPRE) that is activated by binding PPARα to a universal promoter expressed under normal culture conditions. A plasmid in which a firefly luciferase DNA construct was bound was purchased and used.

Dispense CV-1 cells at a constant concentration in the plate incubator and mix the PPAR-α DNA and PPRE with the firefly-luciferase DNA construct and the renilla-luciferase construct in serum-free medium and transfect with lipofectamine. Sean. After transfection, the test substances were treated by diluting each medium to a predetermined concentration, and incubated in a CO 2 incubator at 37 ° C. for a predetermined time. After incubation, the cell eluate was prepared using a reporter lysis buffer, transferred to a micro tube, and centrifuged at 13,000 rpm for 5 minutes to separate the cell residue, thereby obtaining a pure cell eluate.

PPAR alpha activity was determined by the ratio of Renilla luciferase activity to firefly luciferase activity. Therefore, the activities of firefly and lenilla luciferase in the cell disruption solution were measured by luminometer, respectively, and then the firefly luciferase activity value was calculated by dividing by the Renilla luciferase activity value. The firefly / renilla luciferase activity value of the untreated group was converted to 1, and the firefly / renilla luciferase activity value of the test material was quantified by increased activity (fold activity) compared to the untreated group.

Dispense CV-1 cells into a plate incubator and incubate 90% under CO2 5% and 37 ° C. Transfect the plasmids into cells using lipofectamine, and after drug treatment, use the reporter lysis buffer for cell elution. Was prepared. This was transferred to a micro tube and centrifuged at 13,000 rpm for 5 minutes to separate cell residues to obtain pure cell eluate. The PPAR alpha activity of each test substance was mixed in a 1: 1 ratio of luciferase substrate to the cell eluate, the amount of light emitted by the luminometer was measured, and the PPAR alpha activity was compared based on the untreated group. At this time, a positive control group was treated with PPAR activator WY14643 (trade name, pirinixic acid).

As a result, the citronellinic racemic mixture, caffeic acid, (S)-(-)-citronellic acid and (R)-(+)-citronellinic acid treatment groups were all PPAR alpha compared to the untreated group. Activity increased (FIG. 1). In addition, it was confirmed that PPAR alpha activity was increased in a concentration-dependent manner in the case of citronellinic racemic mixture and caffeic acid (FIGS. 2 and 3, respectively). 1: 1: citronelic acid racemic mixture, 2: caffeic acid, 3: (S)-(-)-citronelic acid, 4: (R)-(+)-citronelic acid treatment group .

<1-2> Skin keratin formation promoting ability

Human normal keratinocytes were passaged in medium containing growth factors. Thereafter, the cells were plated in a plate incubator, cultured at 70% to 80% under conditions of 5% CO2 and 37 ° C, and the samples were treated to test epidermal cell differentiation. At this time, 1.2mM CaCl2 treatment group was used as a control group. These were incubated at 37 ° C. for 48 hours, washed with PBS, and the cells were recovered, boiled at 85 ° C. for 15 minutes to elute the cells. After centrifugation at 20,000rpm for 60 minutes to precipitate the cell residue, the protein was quantified using a portion of the supernatant and transferred to a 96-hole plate incubator to measure the absorbance at 430nm conditions. 100% of the sample and no CaCl2 treatment were compared, and the relative degree of epidermal cell differentiation was compared.

As a result, the citronellic racemic mixture, the caffeic acid, the (S)-(-)-citronellic acid and the (R)-(+)-citronellinic acid treatment groups were all keratinocytes, ie epidermis. Enhancement of cell differentiation was found to promote skin keratinocyte formation (FIG. 4). In particular, in the case of citronellic acid racemic mixture and caffeic acid, the formation of keratin was promoted in a concentration-dependent manner (FIGS. 5 and 6, respectively). 4: 1: citronellinic acid racemic mixture, 2: caffeic acid, 3: (S)-(-)-citronelic acid, 4: (R)-(+)-citronelic acid treated group .

<1-3> Expression Enhancement of Involuclin and Transglutaminase-1

Human epidermal cells were passaged in medium containing growth factors. This was dispensed into a plate incubator and incubated at 70% to 80% under CO 2 5% and 37 ° C. and then treated with caffeic acid by concentration. In this case, 1.2 mM CaCl 2 treatment group was used as a control group, and the expression levels of involukrin and transglutaminase-1, which play a decisive role in skin keratin formation, were compared. After incubation for 48 hours at 37 ℃, washed with PBS, lysed cells with RIPA lysis buffer and centrifuged to obtain a cell eluate. Proteins were quantified by Bradford assay and proteins were isolated by electrophoresis. It was transferred to PVDF membrane and reacted with Involuclin and Transglutaminase-1 antibodies, respectively, and developed on the film.

As a result, the expression of involuclin was found to increase in a concentration-dependent manner for caffeic acid compared to the untreated group (FIG. 7), and the expression of transglutaminase-1 was also increased for caffeic acid compared to the untreated group. It was confirmed that the concentration-dependent increase (Fig. 8).

Experimental Example 2 Moisturizing Skin

Human epidermal cells were passaged in medium containing growth factors. This was dispensed into a plate incubator and incubated at 70% to 80% under CO2 5% and 37 ° C. and then treated with caffeic acid by concentration. At this time, 1.2mM CaCl2 was treated as a control to compare the skin moisturizing ability. After incubation for 48 hours at 37 ℃ washed with PBS and RNA was isolated using Trizol reagent. CDNA was synthesized using Reverse Transcription Premix followed by PCR using each primer.

As a result, propyllagrin, a precursor protein of natural moisturizing factor (NMF), responsible for skin moisturizing, It was confirmed that the expression of the gene increases in a concentration-dependent manner to caffeic acid compared to the untreated group (Fig. 9). Propyllagrin is a precursor of filaggrin, which is an important regulatory protein for epidermal homeostasis, which is a component of NMF, which is a moisturizer when it breaks down, and caffeic acid is thought to enhance skin moisturization.

Experimental Example 3 Antioxidant Activity

The antioxidant activity of caffeic acid was confirmed using DPPH free radical scavenging method. 0.1 mM DPPH methanol solution and concentration-specific samples were reacted 1: 1 at 37 ° C. for 30 minutes, and then absorbance at 516 nm was measured to quantify the concentration of DPPH. Free radical scavenging activity was calculated by the following <Formula 1>, and ascorbic acid of the same concentration was used as a positive control.

<Formula 1>

Free radical scavenging (%) = [1- (BC) / A] ㅧ 100

A: absorbance after reacting without adding a sample,

B: Absorbance after adding and reacting with a sample

C absorbance when methanol is substituted for DPPH

As a result, the IC 50 of caffeic acid was 21.6 μM, and it was confirmed that it had an excellent antioxidant power similar to ascorbic acid (IC 50 = 23.5 μM) (FIG. 10).

Experimental Example 4 Skin Irritation Relief

The forearm of both arms of the subject was designated as the test site, and the SLS (Sodium Lauryl sulfate, Sigma) 1.5% solution was closed and closed for 24 hours using an IQ chamber. After removing the patch and waiting in a constant temperature and humidity conditions, the transdermal moisture loss (TEWL: Transepidermal water loss, measuring unit: g / h ㎡) was measured by using a TEWAMETER TM210 (C + K, Germany) at the test site. Transdermal moisture loss is a useful method for measuring skin barrier function and is directly related to the measurement of the water content of keratin worms. Lower TEWL value means better skin condition, and the improvement rate was calculated according to <Formula 2> using the measured values before and after using the sample. At this time, the sample was prepared according to the following Table 1, and after 1, 2, 7 days after using the sample to evaluate the skin irritation relief. At this time, in order to measure the degree of natural healing after damage to the barrier was used as a control group.

<Formula 2>

TEWL improvement (%) = (Measurement of transdermal moisture loss before sample use) Measured value of transdermal moisture loss after sample use / Measured transdermal moisture loss before sample use) x 100

As a result, the skin irritation improvement rate of the experimental groups to which the sample was applied was better than that of the non-coated group, and the skin barrier improvement was improved through the improvement of the skin barrier (Table 2).

Claims (4)

Cosmetic composition for promoting keratin film formation comprising citronellinic acid or a salt thereof as an active ingredient
The method of claim 1,
A cosmetic composition characterized by having epidermal cell differentiation promoting ability, PPAR alpha activity promoting ability, skin barrier strengthening ability or skin stimulating ability.
The method of claim 1,
The citronellinic acid is a cosmetic composition, characterized in that their optical isomers, non-racemic mixtures thereof, or racemic mixtures.
A pharmaceutical composition for promoting keratin film formation comprising citronellinic acid or a pharmaceutically acceptable salt thereof as an active ingredient.

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