KR101326026B1 - Cosmetic Composition Containing Peptide Derivatives - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin itch and / or skin whitening, and more specifically, the cosmetic composition is a peptide derivative (palmitoyl-Lys-Thr-Thr-Lys-Ser) having an inhibitory effect on PAR-2 activity. ) As an active ingredient is characterized by having an excellent skin itch improvement and thereby atopic dermatitis improvement and / or skin whitening effect.

Description

Cosmetic composition containing peptide derivatives {Cosmetic Composition Containing Peptide Derivatives}

The present invention relates to a cosmetic composition for improving skin itch and / or skin whitening, and more specifically, the cosmetic composition is a peptide derivative (palmitoyl-Lys-Thr-Thr-Lys-Ser) having an inhibitory effect on PAR-2 activity. ) As an active ingredient is characterized by having an excellent skin itch improvement and thereby atopic dermatitis improvement and / or skin whitening effect.

Atopic dermatitis is a recurrent chronic dermatitis with severe itching (pruritus). It is estimated that 10 to 20% of the world's population suffers from atopic dermatitis, and the recent increase in airborne allergens caused by environmental pollution The incidence rate continues to rise due to factors such as the following. The cause of atopic dermatitis is not clear to date, but it is estimated to be caused by a combination of genetic predisposition, immune system abnormalities, itching characteristics easily compared to normal people, infection caused by bacterial virus fungus, and emotional and environmental factors. It is becoming.

Conventionally, in order to improve such atopic dermatitis, a lipid complex of ceramide to moisturize and restore the skin barrier is applied (Korean Patent Laid-Open No. 2001-0044622), oral administration of anti-stamine to treat itching, anti-inflammatory For example, a method of applying a steroidal drug for the use has been generally used, and a method of applying oligosaccharides to improve the skin immune status has been proposed (Korean Patent Publication No. 2000-0029754). There are problems such as drowsiness due to oral administration of steroids, antihistamines, immunosuppressants, and the like, lowering immunity and safety against skin due to topical administration, and stability when formulating the composition for external application of the skin.

Atopic dermatitis is especially characterized by severe itching, which can cause scratches, which can lead to bacterial infections and secondary infections caused by external allergens, which in turn activate immune cells such as T-cells and macrophages. Cytokines and histamine and neuropeptides that cause itching are secreted, causing inflammation and itching to scratch again, leading to amplified symptoms of atopic dermatitis.

Therefore, when itching is one of the characteristics of atopic dermatitis, wounds are generated by scratching, and atopic dermatitis can be improved through blocking of atopic dermatitis amplification cycles such as secondary infection, inflammation, and itching caused by these. Therefore, there is an urgent need for the development of substances that have no side effects on the human body and have an itching effect.

Human skin color is also genetically determined by the concentration and distribution of melanin in the skin, but is also influenced by environmental or physiological conditions such as sun ultraviolet rays, fatigue and stress. Melanin is a type of amino acid tyrosine (tyrosine) enzymes called tyrosinase (tyrosinase) acts to convert to dopa (DOPA), dopaquinone (dopaquinone) is produced through a non-enzymatic oxidation reaction. However, although the pathway by which melanin is made is known, it is still unknown in detail the mechanism by which tyrosinase acts, the mechanism that induces melanin synthesis.

On the other hand, by inhibiting the synthesis of melanin pigment, the skin tone can be brightened and the skin whitening can be realized, and the skin hyperpigmentation such as blemishes caused by ultraviolet rays, hormones and heredity, ultraviolet rays, and freckles caused by heredity can be improved.

Thus, conventionally, a substance having an inhibitory activity on tyrosinase such as hydroquinone, ascorbic acid, kojic acid, glutathione, and the like is added to the ointment or cosmetics, so that the skin whitening, blemishes, Although it has been used for the purpose of improving freckles, the whitening effect of hydroquinone and the like is recognized, but the amount of use is extremely limited due to severe skin irritation. In addition, ascorbic acid is easily oxidized, and the cosmetics containing the same cause problems such as discoloration and deodorization, and thiol compounds such as glutathione and cysteine have a characteristic unpleasant odor and have problems with transdermal absorption. In addition, these glycosides and derivatives have a high polarity, so there is a problem to use them as a component of a cosmetic. Also, placenta extracts and the like have a problem that the whitening effect is insufficient. Therefore, there is an urgent need for the development of a material having no side effects and excellent whitening effect.

In order to meet the above requirements, the present invention by developing a substance that inhibits the activity of PAR-2 known to affect the skin itch and skin pigmentation, by applying it as a cosmetic, it has no side effects and safe and itching / Or to provide a cosmetic composition excellent in the skin lightening effect.

An object of the present invention is to provide a PAR-2 activity inhibitor containing palmitoyl-Lys-Thr-Thr-Lys-Ser as an active ingredient.

Another object of the present invention is to provide a cosmetic composition for improving the itch containing palmitoyl-Lys-Thr-Thr-Lys-Ser as an active ingredient.

Another object of the present invention is to provide a cosmetic composition for skin whitening containing palmitoyl-Lys-Thr-Thr-Lys-Ser as an active ingredient.

The cosmetic composition according to the present invention has no side effects on the human body and has an excellent effect to relieve itching, and has excellent effect on improving contact dermatitis, in particular, atopic dermatitis, and also excellent skin whitening effect. Very useful for

Recently, neurophysiological and immunological studies on itching in atopic dermatitis have been actively conducted. In particular, in the itch of atopy patients, protease-activated receptor 2 (PAR-2) is excessively expressed in the lesion area of the atopy patient compared to the non-lesion area, and Trita which activates PAR-2 is also expressed. The amount of trytase has been increased by four times, and atopic dermatitis has been shown to be more sensitive to itching than a normal person when treated with a substance that activates PAR-2. Research has been reported that PAR-2 is deeply involved (J. Neurosci., 2003, 23 (15) 6176-6180). In addition, mice knocked out of the PAR-2 gene have been reported to greatly reduce type 4 contact allergens caused by triggers such as flavors, preservatives, surfactants, etc. [Jpn.J. Pharmacol. 88, 77-84 (2002)]. These findings indicate that PAR-2 is involved in not only atopic dermatitis, but also itch symptoms of contact dermatitis.

PAR-2 is a membrane-bounded receptor present in keratinocytes, mast-cells and neurons that is activated by serine proteases to induce NO and TNF-α secretion, leading to inflammation. It is known to cause itching by promoting the secretion of histamine, substance P (Substance-P) and calcitonin gene-related peptide (CGRP) (J. Invest. Dermatol., 2006 (126), 746- 754). More specifically, activating PAR-2 with trypsin in cultured keratinocytes temporarily increases the amount of intracellular calcium (Cell Growth Differne., 8 (1997), 743-749). PAR-2 activation in sensory neurons increases the intracellular calcium concentration and promotes itching by promoting the secretion of CGRP, substance-P (TRENDS in Pharmacological Sciences, 22 (3), (2001), 146- 152).

In addition, PAR-2 has been reported to be involved in the delivery of mature melanosomes in melanocytes to keratinocytes (Pigment Cell Res. 2001, 14 (4), 236-42). It is expected to block the delivery of melanin and show excellent whitening effect.

As described above, PAR-2 is deeply involved in itching and skin whitening of type 4 contact dermatitis due to fragrances, preservatives, surfactants, etc. as well as itching of atopic dermatitis.

 Therefore, the present inventors conducted a study to develop PAR-2 activity inhibitors for extracts and synthetic compounds of various plants and animals to develop substances having an effect on itching and skin whitening of atopic dermatitis and contact dermatitis. Toil-Lys-Thr-Thr-Lys-Ser was found to have excellent PAR-2 activity inhibitory effect and thereby itching inhibitory effect, atopic dermatitis improvement effect and skin whitening effect was completed the present invention. Peptide derivative palmitoyl-Lys-Thr-Thr-Lys-Ser used in the present invention has already been demonstrated to have an anti-aging effect by promoting collagen synthesis (US6,620,419 B1), which is the same as in the present invention. It is distinguished from the inhibition of PAR-2 activity and the itching inhibition and skin lightening effect.

In the present invention, the peptide derivative palmitoyl-Lys-Thr-Thr-Lys-Ser showed an excellent inhibitory effect of PAR-2 activity inhibiting the effect of trypsin-induced intracellular calcium concentration when treated with trypsin in HaCaT cells. As a result of the preparation of skin external ointment or cosmetic containing the peptide derivative palmitoyl-Lys-Thr-Thr-Lys-Ser and applying it to the affected area of atopic patients, it was found that the effect of suppressing itching and atopic dermatitis was very excellent. appear. That is, the cosmetic composition containing the peptide derivative palmitoyl-Lys-Thr-Thr-Lys-Ser of the present invention has an excellent effect on improving the itch, and thus may have an excellent effect on improving contact dermatitis, in particular, atopic dermatitis. .

In addition, when the peptide derivative palmitoyl-Lys-Thr-Thr-Lys-Ser was applied to the skin of normal humans, the skin whitening effect was very excellent and no adverse effects were observed, which is safe and improves blemishes and freckles. Able to know. Therefore, the cosmetic composition comprising the peptide derivative palmitoyl-Lys-Thr-Thr-Lys-Ser according to the present invention may have an excellent skin whitening effect and thereby a blemish and freckling improvement effect.

In another embodiment, the cosmetic composition comprising the derivative palmitoyl-Lys-Thr-Thr-Lys-Ser of the present invention may be one having an itch improvement effect and a skin whitening effect.

The cosmetic composition according to the present invention may be formulated as an external skin preparation or cosmetics. The external preparation for the skin may include an ointment, a warning agent, a spray, a suspension, an emulsion, a cream, a gel, and the like. The cosmetics may include basic cosmetics, makeup cosmetics, body cosmetics, shaving cosmetics, and the like. Examples of the basic cosmetics include cream, lotion, pack, massage cream, latex, etc. Examples of the makeup cosmetics include foundation, makeup base, lipstick, eyeshadow, eyeliner, mascara, eyebrow pencil, etc. Examples of cosmetics include soaps, liquid detergents, baths, sunscreen creams, sun oils, and the like. Examples of shaving cosmetics include aftershave and shaving creams.

The content of palmitoyl-Lys-Thr-Thr-Lys-Ser contained in the cosmetic composition of the present invention is 0.00001 to 1.0% by weight, preferably 0.0001 to 0.1% by weight based on the total dry weight. At a concentration below 0.00001% by weight, no obvious effect could be expected. At concentrations above 1.0% by weight, there was no apparent effect increase with increasing content.

The cosmetic composition of the present invention, in addition to the active ingredient, all kinds of ingredients usable in ordinary cosmetics, such as perfumes, pigments, fungicides, antioxidants, preservatives, humectants, thickeners, excipients, diluents, inorganic salts and synthetic polymer materials And the like, and the content thereof may be appropriately adjusted according to the purpose and purpose of use of the final product.

Hereinafter, the present invention will be described in detail with reference to Examples. However, the following examples may be modified in many different forms, which are intended to illustrate the present invention, but the scope of protection of the present invention is not limited to the following examples. Examples are provided, but the following examples are merely illustrative of the present invention, and the scope of the present invention is not limited to the following examples.

[ Example ]

Test Example  One: Peptides  derivative Palmy toil - Lys - Thr - Thr - Lys - Ser of PAR -2 inhibitory effect test

Palmitoyl-Lys-Thr-Thr-Lys-Ser (Peptron) was added to the culture medium of HaCaT cells at different concentrations to determine the inhibition of PAR-2 activity by measuring intracellular calcium concentration changes at the cellular level (W. Luo et al. Brain Research 1047 (2005) 159-167).

HaCaT cells are incubated in a thermohygrostat with DMEM growth medium (Dulbecoo's modified eagles medium, Cambrex). In a culture dish containing HaCaT cells, fura-2 / AM was treated at a concentration of 5 μM and placed in cells for 30 to 60 minutes with light blocked at room temperature. At the end of the load, the cells were washed twice with extracellular perfusate and then placed on a microscope. 10 nM Trypsin was perfused at a rate of 1 to 2 ml / min by gravity and loaded for 50 to 100 seconds, followed by gravity with extracellular perfusate. It was washed by perfusion at a rate of 1-2 ml / min. Then perfuse each sample containing 10 nM trypsin and palmitoyl-Lys-Thr-Thr-Lys-Ser at a final concentration of 0.0001% to 0.1% by weight at a rate of 1-2 ml / min by gravity. It was loaded for 50 to 100 seconds.

The changes in fluorescence intensity appearing at this time were analyzed and the ratio of fluorescence intensity emitted at 510 nm when excitation alternately with 340 nm and 380 nm light due to the fluorescence characteristics of fura-2 (F340 / F380) This intracellular ion concentration is reflected. In one experiment, changes in calcium ion concentrations in at least 10 single cells were measured. Final concentrations of palmitoyl-Lys-Thr-Thr-Lys-Ser containing 10nM trypsin using control of intracellular calcium changes (a ratio of fluorescence intensity (F340 / F380)) when 10nM Trypsin alone was used as a control Intracellular calcium concentration change (fluorescence intensity ratio (F340 / F380)) was calculated as the inhibition rate for PAR-2 (%), and the results are summarized in Table 1 below.

Inhibitory Effect of PAR-2 Activity at the Cell Level According to Palmitoyl-Lys-Thr-Thr-Lys-Ser Concentration (Repeat Number: 3) sample Application concentration (%)
Calcium Concentration Change
(F340 / F380) % Inhibition of PAR-2 activity Control group (no addition)
- 0.073 ± 0.002 - Palmitoyl-Lys-Thr-Thr-Lys-Ser Final concentration (0.0001%) 0.174 ± 0.007 62% Final concentration (0.001%) 0.078 ± 0.003 98% Final concentration (0.01%) 0.077 ± 0.004 99% Final concentration (0.1%) 0.075 ± 0.006 100% Trypsin Final concentration (10nM)
0.337 ± 0.004
-

As can be seen from the results of Table 1, it can be seen that palmitoyl-Lys-Thr-Thr-Lys-Ser strongly inhibits PAR-2 activation by trypsin in HaCaT cells. In addition, palmitoyl-Lys-Thr-Thr-Lys-Se exhibits a very potent inhibitory effect on PAR-2 activity at 0.0001% by weight without showing cytotoxicity at 0.1% by weight, resulting in atopic dermatitis by inhibiting itching of atopic dermatitis. It has been confirmed that it can be very usefully used as a dermatitis improving agent and skin lightening agent.

The preparation example of the cosmetic containing the peptide derivative, palmitoyl-Lys-Thr-Thr-Lys-Ser is shown by the preparation example below.

Manufacturing example 1 and Comparative Example  1: Preparation of skin external ointment

The composition was prepared as shown in Table 2 below to prepare an external skin ointment containing palmitoyl-Lys-Thr-Thr-Lys-Ser.

Furtherance Production Example 1 (% by weight) Comparative Example 1 (% by weight) Palmitoyl-Lys-Thr-Thr-Lys-Ser
-Diethyl sebacate
-Payment
Polyoxyethylene oleyl ether
Phosphate
Sodium benzoate
-vaseline 0.1
8
5
6

Suitable amount
to 100 -
8
5
6

Suitable amount
to 100

Manufacturing example 2 and Comparative Example  2: preparation of cream

The composition was prepared as shown in Table 3 below to prepare a cosmetic cream containing palmitoyl-Lys-Thr-Thr-Lys-Ser.

Furtherance Preparation Example 2 (% by weight) Comparative Example 2 (% by weight) Palmitoyl-Lys-Thr-Thr-Lys-Ser
Stearic acid
Cetanol
Potassium hydroxide
-glycerin
Propylene glycol
-antiseptic
-incense
-Purified water 0.01
15.0
1.0
0.7
5.0
3.0
Suitable amount
Suitable amount
to 100 -
15.0
1.0
0.7
5.0
3.0
Suitable amount
Suitable amount
to 100

Manufacturing example 3 and Comparative Example  3: Manufacture of Soft Cosmetics

The flexible cosmetics containing palmitoyl-Lys-Thr-Thr-Lys-Ser were prepared as shown in Table 4 below.

Furtherance Production Example 3 (% by weight) Comparative Example 3 (wt%) Palmitoyl-Lys-Thr-Thr-Lys-Ser-
-ethanol
Polylauric acid polyoxyethylene sorbent
Paraoxybenzoic acid methyl
-glycerin
-1,3-butylene glycol
-incense
Pigment
-Purified water 0.0001
10.0
1.0
0.2
5.0
6.0
Suitable amount
Suitable amount
to 100 -
10.0
1.0
0.2
5.0
6.0
Suitable amount
Suitable amount
to 100

Manufacturing example 4 and Comparative Example  4: Preparation of Nutrients

The nutritional longevity containing Palmitoyl-Lys-Thr-Thr-Lys-Ser was prepared as shown in Table 5 below.

Furtherance Production Example 4 (% by weight) Comparative Example 4 (wt%) Palmitoyl-Lys-Thr-Thr-Lys-Ser
Waselin
Sesquioleate sorbitan
Polyoxyethylene oleylethyl
Paraoxybenzoic acid methyl
Propylene glycol
-ethanol
Carboxy vinyl polymer
- potassium hydroxide potassium
Pigment
-incense
-Purified water 0.0005
2.0
0.8
1.2
Suitable amount
5.0
3.2
18.0
0.1
Suitable amount
Suitable amount
to 100 -
2.0
0.8
1.2
Suitable amount
5.0
3.2
18.0
0.1
Suitable amount
Suitable amount
to 100

Manufacturing example 5 and Comparative Example  5: Manufacture of pack

A pack containing palmitoyl-Lys-Thr-Thr-Lys-Ser was prepared as shown in Table 6 below.

Furtherance Preparation Example 5 (% by weight) Comparative Example 5 (wt%) Palmitoyl-Lys-Thr-Thr-Lys-Ser
-glycerin
Propylene glycol
Polyvinyl alcohol
-ethanol
-Polyoxyethylene oleyl ethyl
Paraoxybenzoic acid methyl
-incense
Pigment
-Purified water 0.001
5.0
4.0
15.0
8.0

1.0
0.2
Suitable amount
Suitable amount
to 100 -
5.0
4.0
15.0
0

1.0
0.2
Suitable amount
Suitable amount
to 100

Manufacturing example 6 and Comparative Example  6

The composition was prepared as shown in Table 7 below to prepare an essence containing palmitoyl-Lys-Thr-Thr-Lys-Ser.

Composition Production Example 6 (% by weight) Comparative Example 6 (wt%) Palmitoyl-Lys-Thr-Thr-Lys-Ser
Propylene glycol
-glycerin
Sodium hyaluronate aqueous solution (1%)
-ethanol
Polyoxyethylene Cured Castor Oil
Paraoxybenzoic acid methyl
-incense
-Purified water 0.005
10.0
10.0
5.0
5.0
1.0
0.1
Suitable amount
to 100 -
10.0
10.0
5.0
5.0
1.0
0.1
Suitable amount
to 100

Experimental Example  2

A sample of palmitoyl-Lys-Thr-Thr-Lys-Ser and palmitoyl-Lys- prepared according to Preparation Example 2 and Comparative Example 2 were selected from 20 patients with severe itching among patients diagnosed with atopic dermatitis. Thr-Thr-Lys-Ser-free samples were applied to the affected areas of both arms twice daily for 4 weeks. Then, the degree of itching and the degree of atopic dermatitis were determined by questioning and visual observation, and the improvement of the itching and atopic dermatitis improvement of the sample prepared by the preparation example compared with the sample prepared by the comparative example was markedly effective, effective, and no difference. Was evaluated in three steps, and the results are shown in Table 8 below.

Improvement of Itching and Atopic Dermatitis in Cosmetics Containing Palmitoyl-Lys-Thr-Thr-Lys-Ser Test Items Distinct effect (n.) Effective (persons) No difference (persons) Itching Improvement 4 12 4 Atopic Dermatitis Improvement 2 12 6

As can be seen from the results of Table 8 above, the sample containing palmitoyl-Lys-Thr-Thr-Lys-Ser prepared according to Preparation Example 2 improved the itch and atopic dermatitis out of 20 subjects, respectively. It was excellent at 80% and 70%, and did not show any side effects in the skin, indicating that palmitoyl-Lys-Thr-Thr-Lys-Ser is a safe and effective cosmetic for improving itching and atopic dermatitis.

Test Example  3

Twenty healthy males and females were selected and aluminium foil with 6 holes of 7 mm diameter was duplexed into the lower forearm of both arms, and 60mJ / cm2 light was irradiated using ORIEL solar simulator 1000W at a distance of 10cm from the arm. The irradiation site was washed well with 70% ethanol aqueous solution before irradiation. Samples containing palmitoyl-Lys-Thr-Thr-Lys-Ser and palmitoyl-Lys prepared according to Preparation Examples 1 to 6 and Comparative Examples 1 to 6 twice a day for 3 weeks after irradiation from 3 days before irradiation. Samples containing no -Thr-Thr-Lys-Ser were applied in pairs on the same row (for the formulation of packs of Preparation 5 and Comparative Example 5 and removed 15 minutes after application). The pigmentation degree of the preparation example and the comparative example was judged visually about each, and the pigmentation inhibition degree of the sample manufactured by the preparation example compared with the sample manufactured by the comparative example was markedly effective, effective, and no difference 3 Evaluated in steps, the results are shown in Table 9 below.

Pigmentation Inhibitory Effects of Cosmetics Containing Palmitoyl-Lys-Thr-Thr-Lys-Ser Experimental material Distinct effect (n.) Effective (persons) No difference (persons) Production Example 1 5 9 6 Production Example 2 4 8 8 Production Example 3 2 9 9 Production Example 4 2 8 10 Production Example 5 2 9 9 Production Example 6 3 10 7

As can be seen from the results in Table 9, the palmitoyl-Lys-Thr-Thr-Lys-Ser-containing sample prepared according to Preparation Example 1-6 showed a whitening effect on at least 10 of 20 subjects. In particular, the comparative experiments of Preparation Example 1 and Comparative Example 1 to which a large amount of palmitoyl-Lys-Thr-Thr-Lys-Ser were applied showed that the pigmentation inhibitory effect was found in 70% of the subjects, and any side effects in the skin were observed. It does not appear, palmitoyl-Lys-Thr-Thr-Lys-Ser is a safe and effective very effective for improving skin or freckles or skin whitening cosmetics.

Claims (2)

A cosmetic composition for skin whitening containing palmitoyl-Lys-Thr-Thr-Lys-Ser as an active ingredient.
The cosmetic composition according to claim 1, wherein the content of palmitoyl-Lys-Thr-Thr-Lys-Ser is 0.00001 to 1.0% by weight based on the dry weight of the total composition.