KR20210150839A - Cosmetic composition effective for skin improvement containing palmitoyl tripeptide compound - Google Patents

Abstract

The present invention relates to a cosmetic composition effective in skin improvement, containing a palmitoyl tripeptide compound. The cosmetic composition according to the present invention contains the compound, thereby having a skin aging inhibitory effect based on excellent wrinkle alleviation and skin elasticity improvement. The compound also provides an excellent effect in skin regeneration by promoting collagen production in damaged skin tissues.

Description

Cosmetic composition effective for skin improvement containing palmitoyl tripeptide compound

The present invention relates to a novel peptide derivative compound and a cosmetic composition for inhibiting skin aging containing the compound.

The skin functions to protect the human body from various external threats and is prone to aging because it is always in contact with various external stimuli. As the skin tissue ages, the ability to produce fibers and matrix decreases, and accordingly, when the amount of matrix decreases, the thickness of the skin becomes thinner and wrinkles are formed by lowering the elasticity of the skin. Various compounds are known to be useful for improving skin appearance and physiological function, including fine lines, wrinkles, aging, or other symptoms associated with photodamaged skin. For example, skin improvement effects such as vitamin C and retinoids have been well known, and cosmetic compositions containing them are being sold in the global market and are in the spotlight. However, the compositions for skin improvement according to the prior art have been reported to have a side effect equal to the skin improvement effect, that is, problems causing stability problems during storage and skin irritation.

Peptides are examples of compounds having a skin improvement effect while having relatively few side effects of skin irritation. As a major component of all proteins, peptides have the advantage of activating skin regeneration without adverse effects on the skin. According to a recently published study, cosmetics containing peptides or peptide derivatives have a very fast skin improvement effect compared to retinol, which is generally used as a conventional wrinkle improvement agent, and have no side effects even when used in large amounts, so it is safe even for sensitive areas such as the eyes. and can be used.

Currently, various peptide derivatives used in wrinkle-improving cosmetics are known. Examples of such peptide derivatives include tripeptide derivatives such as Palm-Gly-His-Lys, tetrapeptide derivatives such as Palm-Gly-Gln-Pro-Arg, pentapeptides, and hexapeptides, and cosmetic products containing them as main components Some cosmetic products made of the composition are commercially available. The peptide derivative components play a role in ultimately removing skin wrinkles and regenerating the skin by performing a collagen activation function in the skin soft tissue.

Currently, the mainstream of research on cosmetic compositions for skin improvement is focused on finding novel compounds that activate collagen. The main amino acids constituting collagen are glycine, proline, and hydroxyproline, and it is characterized by a high content of hydroxyproline, which is not present in other proteins. Accordingly, Gly-Pro-Hyp composed of the three amino acids is known as a peptide representing collagen.

According to several papers (Acta Neurobiol. Exp. 1994, 54:33-38, Acta Neurobiol. Exp. 1992, 52:223-232), the peptide (Gly-Pro-Hyp) representing collagen is applied to the skin It is confirmed that it helps a lot in wrinkle improvement. Currently, Gly-Pro-Hyp is used as a component for collagen activation and skin wrinkle improvement in various products in Japan and the United States. However, since Gly-Pro-Hyp is made of pure hydrophilic ingredients, it is difficult to pass through the skin barrier (ceramide layer).

As described above, various peptides having the same skin improvement effect and cosmetic compositions containing derivatives of the peptides have been reported, but cosmetic compositions that satisfy various needs such as wrinkle improvement, moisturizing, anti-aging, and skin regeneration with few side effects are reported. The composition has not yet been reported.

The present invention is to overcome the problems of the prior art as described above. In order to overcome the difficulty of penetrating the skin barrier of Gly-Pro-Hyp, which is a hydrophilic peptide, a hydrophobic palmitoyl group is introduced into the Gly-Pro-Hyp. To provide a novel palmitoyl tripeptide derivative having a hydrophilic-lipophilic structure. In addition, to provide a method for preparing the novel palmitoyl tripeptide derivative and a cosmetic composition containing the same.

According to one aspect of the present invention, a novel palmitoyl represented by the following structural formula I or palmitoyl-glycinyl-prolinyl-hydroxyproline (Palmitoyl-Glycinyl-Prolinyl-Hydroxyproline; Palm-Gly-Pro-Hyp) Tripeptide compounds are provided.

[Structural Formula I]

According to another aspect of the present invention, using 2-(4-nitrophenyl)sulfonylethoxycarbonyl-amino acid (Nsc-amino acid) as a reactant, glycinyl-pe Synthesis of rollinyl-hydroxyprolinyl-resin (Gly-Pro-Hyp-resin), palmitoyl-glycin by introducing a palmitoyl group into the glycinyl-prolinyl-hydroxyprolinyl-resin Synthesis of cinyl-prolinyl-hydroxyproline-resin (Palm-Gly-Pro-Hyp-resin), and a resin from the palmitoyl-glycinyl-prolinyl-hydroxyprolinyl-resin And it provides a method for preparing a palmitoyl tripeptide compound (Palm-Gly-Pro-Hyp) comprising the step of removing the side chain protecting group.

According to another aspect of the present invention, there is provided a cosmetic composition for wrinkle improvement and anti-aging containing the palmitoyl tripeptide compound.

Alternatively, the cosmetic composition may be a softening lotion (skin), a nourishing lotion, or a nourishing cream.

In another method, the content of the palmitoyl tripeptide compound in the cosmetic composition may be 0.0001 to 0.1% by weight.

The cosmetic composition effective for skin improvement containing the palmitoyl tripeptide compound according to the present invention has excellent skin aging, wrinkle improvement, skin elasticity and skin penetration effects, as well as excellent long-term storage stability, skin safety, and ease of use. provide an improvement effect.

1 shows the results of the MTT assay for the palmitoyl tripeptide compound (Palm-Gly-Pro-Hyp) according to the present invention.
Figure 2 shows the measurement result of the collagen synthesis improvement effect of the palmitoyl tripeptide compound (Palm-Gly-Pro-Hyp) according to the present invention.
3 shows the measurement results for each week of the skin wrinkle improvement evaluation test of the cosmetic composition.
Figure 4 shows the strain-time curve of the skin elasticity improvement evaluation test of the cosmetic composition.
5 shows the measurement results for each week of the skin elasticity improvement evaluation test of the cosmetic composition.
6 is a skin lifting measurement photograph of the skin lifting improvement evaluation test of the cosmetic composition.
7 shows the measurement results for each week of the skin lifting improvement evaluation test of the cosmetic composition.

The present invention provides a cosmetic composition effective for skin improvement containing a palmitoyl tripeptide compound.

The novel peptide preparation method of the present invention is 2-(4-nitrophenyl)sulfonylethoxycarbonyl

[2-(4-nitrophenyl)sulfonylethoxycarbonyl]-amino acid (hereinafter referred to as Nsc-amino acid) to prepare palmitoyl tripeptide. Step, 2) a step of introducing a palmitoyl group, 3) a step of removing the resin and the side chain protecting group.

The Nsc-amino acid used in the synthesis method according to the present application is more stable than the conventional 9-fluorenyl methoxycarbonyl (Fmoc)-amino acid, and thus has the advantage of being easier to handle at room temperature. In addition, Nsc-amino acids have an advantage in that the rate of decomposition in DMF, which is a solvent generally used for peptide synthesis, is considerably slower than that of Fmoc, and the Nsc group can be removed under much more relaxed conditions. Accordingly, the synthesis method using Nsc-amino acids can be very advantageously applied to a conventional peptide preparation method in which an amino acid mixture is dissolved in a solvent in advance and a peptide is synthesized using a synthesis reactor, and can also occur due to damage to amino acids before the reaction. Various side reactions can also be reduced. In addition, since Nsc-amino acid is cheaper than Fmoc-amino acid, it is more economical than the conventional manufacturing method using the Fmoc.

The present invention will be described in more detail as follows. (1) A novel palmitoyl tripeptide, Palm-Gly-Pro-Hyp, is synthesized using a peptide synthesis method using Nsc-amino acids. (2) An aqueous solution of the palmitoyl tripeptide is added to the cosmetic composition in an amount of 0.0001 to 0.1% by dry weight, preferably about 0.01%, and the effect is measured.

The examples described below are for explaining the content of the present invention in detail, but the content of the present invention is not limited thereto.

<Example>

The following examples are intended to further describe and explain the embodiments within the scope of the present invention. This is merely illustrative of the present invention and not intended to be limiting, and various modifications may be made without departing from the spirit and scope of the present invention. Abbreviations used in the present specification and examples are as follows.

Nsc: 2- (4-nitrophenyl) sulfonylethoxycarbonyl

Fmoc: 9-fluorenylmethoxycarbonyl

Gly: glycine

Pro: proline

Hyp: hydroxyproline

Palm: Palmitoyl

DIPEA: diisopropylethylamine

DMF: N,N-dimethylformamide

MC: methylene chloride

HOBT: 1-hydroxybenzotriazole

HBTU: 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate

TFA: trifluoroacetic acid

TIS: triisopropylsilane

tBu: tert-butyl

<Example 1> Synthesis of Palm-Gly-Pro-Hyp

Palm-Gly-Pro-Hyp, a novel peptide derivative according to the present invention, was synthesized by a solid-phase peptide synthesis method, and the synthesized peptide derivative was analyzed using HPLC and Maldi-Tof mass.

(a) Introduction of Hyp: 2-Chlorotrityl chloride resin (CTL resin, Bead TechTM, 1% DVB crosslinked, 100-200 mesh; 1.3 mmol/g) 7.7 g (10 mmol) in a reaction vessel was added, and 110 ml of MC was added thereto, followed by stirring for 3 minutes. After removing the solution, 110 ml of DMF was added, stirred for 3 minutes, and the solvent was removed again. Then, 5.33 g (1.2eq) of Nsc-Hyp(tBu)-OH, 50 mL of DMF, and 7 mL (4eq) of DIPEA were added and reacted for 1 hour, washed, and methanol and DIPEA (2:1) were dissolved in MC and reacted for 10 minutes. After washing, it was washed with an excess of MC/DMF (1:1). Hyp-resin was obtained by deprotecting the Nsc group with 50 mL of DMF solution containing 20% piperidine and washing with DMF 3 times. At this time, the dose of bound amino acid per g of resin was 0.64 mmol.

(b) Introduction of Pro: 7.5 g (2eq) of Nsc-Pro-OH, 2.7 g (2eq) of HOBt, and 7.6 g (2eq) of HBTU were dissolved in DMF to Hyp-resin, and then 7 mL (4eq) of DIPEA was added. and reacted. The completion of the reaction was confirmed by the Kaiser test, and the deprotection of the Nsc group after amino acid condensation was reacted twice with 50 mL of DMF solution containing 20% piperidine for 20 minutes each, followed by washing to obtain Pro-Hyp-resin.

(c) Introduction of Gly: 6.6 g (2eq) of Nsc-Gly-OH, 2.7 g (2eq) of HOBt, and 7.6 g (2eq) of HBTU were dissolved in DMF to the Pro-Hyp-resin, respectively, and then DIPEA 7mL ( 4eq) ) was added to react. The completion of the reaction was confirmed by the Kaiser test, and the deprotection of the Nsc group after amino acid condensation was reacted twice with 50 mL of DMF solution containing 20% piperidine for 20 minutes each, followed by washing to obtain Gly-Pro-Hyp-resin.

(d) Introduction of Palmitoyl group: After adding DMF 60mL to the Gly-Pro-Hyp-resin, 6.4mL (2eq) of Palmitoyl chloride and 5.2mL (3eq) of DIPEA were added to the reaction. The completion of the reaction was confirmed by the Kaiser test, and when the reaction was completed, it was washed thoroughly with DMF and MC and dried.

(e) Emission: 70 mL of TFA:TIS:H2O (95:2.5:2.5) was added to the Palm-Gly-Pro-Hyp-resin, and dispersed in 200 mL of diethyl ether, filtered and dried to obtain a purity of 74.4% and a yield of 82 % crude Palm-Gly-Pro-Hyp was obtained. HPLC results are as follows: Rt 31.2 (0-100% 0.1% TFA in ACN for 40min, XTerra ^TM RP ₁₈ , 5㎛, 4.6×250mm)

f) Purification: The crude Palm-Gly-Pro-Hyp was purified by ID 5cm preparative LC and freeze-dried to obtain 2.69 g of a powder of a novel peptide derivative (Palm-Gly-Pro-Hyp). Its purity was 99.1%, and it was found that the molecular weight was 524.40 (theoretical value: 523.71) using a molecular weight meter.

<Example 2> Preparation of aqueous solution of Palm-Gly-Pro-Hyp

In order to add the peptide derivative synthesized in Example 1 to the cosmetic formulation, it was dissolved in an aqueous solution at a concentration of 1000 ppm by the following method.

After dissolving 0.1 g of the Palm-Gly-Pro-Hyp in 12 g of butylene glycol at room temperature, add 1 g of solubilizant LRI, 5 g of ethanol, and 0.4 g of Phenonip, add 81.4 g of purified water, stir well, and use triethanolamine to pH was set to 5.3.

<Example 3> Formulation of flexible lotion (skin)

Table 1 describes a formulation example of a softening lotion (skin) containing palmitoyl tripeptide.

Table 1

<Example 4> Formulation of nutritional lotion

Table 2 describes a formulation example of a nutritional lotion containing palmitoyl tripeptide.

Table 2

<Example 5> Formulation of nourishing cream

Table 3 describes a formulation example of a nutritional cream containing palmitoyl tripeptide.

Table 3

Common cell culture includes monolayer culture, in which cells are attached to the bottom of a culture flask, and suspended culture, in which cells are suspended and grown in a cell culture medium. Among them, the culture method frequently used for skin-related effect testing is the monolayer culture method. This is because the momolayer culture method deals with cells in which fibroblasts, keratinocytes, or endocrine cells related to skin tests are attached and grown. The efficacy evaluation test using the monolayer culture method can know the test result relatively quickly and can be used relatively easily for screening a large amount of sample because the amount of sample required is small. In addition, the burden on the test cost is relatively small. The disadvantages of the monolayer culture method are that only water-soluble substances soluble in the medium can be tested, the difference from the test results in actual skin tissue is relatively large, and since it is directly applied to cells, a buffer for toxicity that there is no

Therefore, recently, in order to compensate for the shortcomings of the monolayer culture method and to obtain a result closer to the actual skin, a 3D artificial culture method in which several related cells constituting the actual skin are cultured together rather than a monolayer culture of specific target cells have been developed. and is being used. The artificial 3D cell culture is also widely applied to study interactions between cells (signal transduction between cells, material transfer). Examples of the artificial 3D cell culture include a method of culturing by additionally planting keratinocytes in a D.E (Dermal Equivalent) culture system, which is an artificial dermis.

According to the 3D experiment, after culturing the artificial dermis for a certain period of time, a culture system is made by dispensing keratinocytes on a collagen sheet. The dispensed keratinocytes make a keratinized barrier on the collagen sheet layer, and some of them migrate to the collagen sheet layer. In this case, thanks to the keratinized barrier, the damping ability against toxicity is high in the efficacy test using the finished product as it is or using high-concentration additives. Therefore, the culture system according to the 3D experiment can be effectively used for testing protein synthesis ability, cell proliferation ability, cell metabolism ability, and analysis of other immune substances. As the media required for culture, a medium such as DMEM media for fibroblast and keratinocyte SFM for keratinocytes is used.

The manufacturing and evaluation method of the 3D culture system is as follows;

1. Artificial dermal D.E (Dermal Equivalent) culture system: Create artificial dermal fluid using collagen matrix and fibroblast and put it in Transwell to make artificial dermis.

2. The keratinocytes are cultured on the culture system cultured for about 1 week to form a stratum corneum.

3. After applying the raw material or cosmetic composition prepared according to the present invention on the artificial skin in which the stratum corneum is sufficiently formed, stimulation (metabolic activity, collagen biosynthesis, anti-inflammatory, etc.) is applied or the sample is processed according to various experimental purposes Measure the results.

<Example 6> Fibroblast proliferation effect of Palm-Gly-Pro-Hyp

In order to examine the fibroblast proliferation effect of the peptide derivative according to the present invention, MTT assay was performed on the Palm-Gly-Pro-Hyp (Sample A) prepared by the method according to Example 1. As a comparative experiment, Gly-Pro-Hyp (Sample B), a peptide into which a palmitoyl group was not introduced, was used.

In order to measure 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), MTT was treated on the artificial skin, and MTT was sufficiently permeated into the cells in the artificial skin, and then applied to the artificial skin. DMSO was administered at 2ml each and after MTT extraction, absorbance was measured at 560 nm using an ELISA reader.

Table 4 shows the MTT assay results for the peptide compound (Palm-Gly-Pro-Hyp) according to the present invention.

Table 4

1 shows the MTT assay results for the peptide compound (Palm-Gly-Pro-Hyp) according to the present invention.

Referring to Table 4 and FIG. 1, it can be seen that the peptide derivative (Palm-Gly-Pro-Hyp) prepared by the method according to Example 1 exhibits an excellent fibroblast proliferation effect compared to the control compound Gly-Pro-Hyp. can

<Example 7> Collagen synthesis improvement effect of Palm-Gly-Pro-Hyp

In order to examine the collagen synthesis improvement effect of the peptide derivative according to the present invention, a Pro-collagen Type I C-peptide (PIP) quantification kit was used for Palm-Gly-Pro-Hyp prepared by the method according to Example 1. Thus, the collagen synthesis improvement effect was measured.

Collagen is the largest structure constituting the ECM (Extra Cellular Matrix; extracellular matrix). Inhibition of collagen biosynthesis and degradation is closely related to the health of ECM. Various types of collagens (Types I, II, III, IV, and V) are synthesized from precursor molecules called procollagens.

These procollagens additionally contain additional peptide sequences called pro-peptides at each of the C-terminal and N-terminal ends of the amino acid chain. The pro-peptides allow pro-collagen molecules to be well wound into a tertiary helix structure and form well into the endoplasmic reticulum. The pro-peptides are cut out when collagen is secreted, and then collagen of the tertiary helix structure is present in the ECM. Therefore, quantification of the amount of pro-peptides can be a reliable standard for quantifying the amount of collagen we are targeting.

The pro-collagen Type I C-peptide (PIP) quantification kit used in the test was manufactured by TAKARA, and the content of pro-peptides in the artificial skin culture medium was evaluated according to the following method.

(a) Put 100 microliters of antibody-POD conjugate into the micro-well. Immediately thereafter, 20 microliters of sample or standard material are added as appropriate for each test purpose.

(b) Cover the microplate with a seal and leave it at 37°C for 3 hours.

(c) After incubation, discard all liquid in each well and wash 4 times using a plate washer.

(d) Substrate treatment: 100 microliters of the substrate solution is put into each well, left at room temperature for 15 minutes, and then 100 microliters of the reaction stop solution is added to each well to stop the substrate reaction.

(e) Measure absorbance at 450 nm.

Table 5 shows the measurement results of the collagen synthesis improvement effect of the peptide derivative compound (Palm-Gly-Pro-Hyp) according to the present invention.

Table 5

2 shows the measurement results of the collagen synthesis improvement effect of the peptide derivative compound (Palm-Gly-Pro-Hyp) according to the present invention.

Referring to Table 5 and FIG. 2, it can be seen that Palm-Gly-Pro-Hyp, a peptide derivative compound prepared by the method according to Example 1, exhibits superior collagen synthesis effect compared to the control compound, Gly-Pro-Hyp. have.

<Example 8> Safety test of cosmetic composition (human patch test)

In order to determine the degree of skin irritation of the cosmetic composition containing the peptide derivative according to the present invention, a skin patch test was performed on the cosmetic composition prepared according to Examples 3 to 5. A total of 15 subjects were selected as subjects aged 20 to 50 years without skin disease at the test site. First, the upper arm, which is the patch site, is wiped with 70% ethanol and dried, and then vaseline is used as a blank using a fin chamber, the softening lotion of Example 3, the nourishing lotion of Example 4, or the Example Each of the nourishing creams of 5 was added in an amount of 0.02 g. Each of the above cosmetic compositions was applied to the upper arm of the human body for 24 hours, and skin reactions were tested within 1 hour after the patch was removed, and then tested again the next day (48 hours). The skin reaction was determined according to the test standards in Table 6, and the evaluation of the skin irritation induction potential was quantified by the average reaction rate calculated from the following formula.

Average response rate (%) = [Σ (number of response subjects after 24 hours × score) + Σ (number of response subjects after 48 hours × score)] × 100 / (total number of tests × maximum score × total number of subjects)

Table 6 describes the inspection standards for the human patch test.

Table 6

The average reaction rate was calculated as follows.

[(15 persons × 0 points + 0 persons × 1 point + 0 persons × 2 points + 0 persons × 3 points + 0 persons × 4 points) + (15 persons × 0 points + 0 persons × 1 point + 0 persons × 2) Points+0 people×3 points+0 people×4 points)]×100/(2 times×4 points×15 people)= 0%

Table 7 shows the skin patch test results for the cosmetic compositions according to Examples 3 to 5.

Table 7

Referring to Table 7, it was confirmed that all of the compositions according to Examples 3 to 5 fell within the non-irritating range and were safe materials for the human body.

<Example 9> Stability test of cosmetic composition

In order to investigate the degree of stability of the cosmetic composition containing the peptide derivative according to the present invention, stability tests were performed on the formulations according to Examples 3 to 5. Specifically, after standing in a thermostat at room temperature (25°C), refrigeration (4°C), or high temperature (40°C) for 3 months, respectively, the change in appearance was observed, and depending on the degree of separation of the formulation, '0' (stable), '±' (slightly unstable) and '+' (unstable) were indicated.

Table 8 shows the stability test results for the formulation examples according to Examples 3 to 5.

Table 8

Referring to Table 8, it was confirmed that the cosmetic compositions of Examples 3 to 5 existed as stable materials at room temperature, refrigeration, and high temperature.

<Example 10> Skin wrinkle improvement evaluation test of cosmetic composition

In order to find out the degree of improvement in skin wrinkles of the composition containing the peptide derivative according to the present invention, clinical efficacy evaluation was performed on 15 healthy women over 30 years of age for the cosmetic compositions of Examples 4 and 5.

The wrinkle improvement effect according to the use of the product was analyzed using a visiometer. To this end, at 0 weeks, 4 weeks, and 8 weeks, imitation plates of the designated eye area were prepared. The simulation plate was manufactured under constant temperature and humidity (22±2° C., 40-60% humidity) conditions. The subjects washed their face with the face wash provided after the visit and were allowed to rest in a constant temperature and humidity (22±2℃, 40% to 60% humidity) condition for 30 minutes. Then, a simulated plate was prepared using silicone solutions B (base) and C (catalyst).

The prepared replica was analyzed using the software of the Visiometer (Skin-Visiometer SV 600, Courage & Khazaha, Germany). The analysis of the replica plate is performed by spectroscopy, and specifically, the intensity of light generated while light emitted from an artificial light source passes through a silicone material is analyzed by Lambert & Beer's Law to measure the improvement of skin wrinkles.

The parameter measured by the visiometer is denoted by R2, which represents Maximum Roughness. As the value of the above variable decreases, skin wrinkles are improved and the depth of wrinkles is lowered, and the unit is an arbitrary unit.

Table 9 shows the skin wrinkle improvement rate (%) in the skin wrinkle improvement evaluation test of the cosmetic composition according to the present invention.

Table 9

*; p<0.05

3 shows the measurement results for each week of the skin wrinkle improvement evaluation test of the cosmetic composition.

Referring to Table 9 and FIG. 3 , it was confirmed that the cosmetic composition helped improve skin wrinkles at a statistically significant level (p<0.05) in the evaluation after 8 weeks of use.

<Example 11> Skin elasticity improvement evaluation test of cosmetic composition

In order to find out the degree of improvement in skin elasticity of the cosmetic composition according to the present invention, clinical efficacy evaluation was performed on 15 healthy women over 30 years of age for the cosmetic compositions of Examples 4 and 5.

Skin elasticity was measured using the Cutometer MPA580 (Courage and Khazaka, Koln, Germany) to measure the elasticity of the eyelid area of the face. The entire subject's face was washed with water and then waited for 30 minutes in a constant temperature and humidity room. At this time, constant temperature and humidity conditions were maintained. The measuring device uses the principle that the skin returns to its original shape when the negative pressure is removed after the skin is sucked into the probe by applying a continuous negative pressure during the measurement time.

As a specific measurement method, a 2 mm diameter probe connected to the measuring device was attached to the subject's hair-removed skin and measured in a non-invasive manner. When negative pressure is applied to the skin surface through the inlet of the probe, the inside thereof is changed to a vacuum state. At this time, the skin surface is pulled into the entrance of the probe. (Maximum 3.0 mm) The length of the pulled skin is measured by an optical system that can pass through the skin. The light used at this time is infrared, and it measures how much the skin is pulled into the probe through the ratio of the intensity of the light that is reduced while passing through the skin and the intensity of the light that does not pass through the skin. The position of the skin close to the pulled skin is fixed by the outer protective ring of the probe. At this time, the pressure applied to the adjacent skin was 50 g/cm ² or 2×10 ³ N/m ² . As the measurement condition, mode 1 was used. Mode 1 condition consisted of a constant sound pressure of 450 mbar, suction time of 2 seconds, and relaxation time of 2 seconds. After three consecutive measurements, the arithmetic average was obtained to obtain the measurement results.

Figure 4 shows the strain-time curve of the skin elasticity improvement evaluation test of the cosmetic composition.

The measured variable used is gross elasticity R2 = Ua/Uf, which means that the closer to 1.0, the more elastic curve is formed.

Table 10 shows the skin elasticity improvement rate (%).

Table 10

*; p<0.05

5 shows the measurement results for each week of the skin elasticity improvement evaluation test of the cosmetic composition.

Referring to Table 10 and Figure 5, it was confirmed that the cosmetic composition according to the present invention helps to improve skin elasticity at a statistically significant level (p<0.05) after 8 weeks of application to the subjects.

<Example 12> Skin lifting improvement evaluation test of cosmetic composition

In order to find out the degree of skin lifting improvement of the cosmetic composition according to the present invention, clinical efficacy evaluation was performed on 15 healthy women over 30 years of age for the cosmetic compositions of Examples 4 and 5.

In order to measure the skin lifting effect of the test product after 30 minutes in a constant temperature and humidity room, an image is obtained using a measuring device on the subject's face. The measuring device is a measuring device that takes a picture under the same conditions as the brightness of indoor lighting using a measuring device and lighting set to which the Moire method is applied and a high-resolution digital camera (Olympus E-330, Japan). In this experiment, for standardization, measurement conditions (measurement direction and location) were fixed.

In particular, among the test areas of the subject, the area around the mouth where the sagging of the skin can be observed was selected as the test site.

As a specific analysis method, in the image taken through the measuring device, a horizontal line is drawn at the end of the lips, a vertical intersection (tangent) of the contour line that meets the end of the lips is drawn, and the angle (R) they make is calculated using an image analysis program (ImagePro, USA) for analysis. The Moire image criterion indicates that the more the contour line is parallel to the corner of the mouth, the better the skin elasticity, and the lower the contour line, the worse the skin elasticity. In this case, the more severe the sagging of the skin around the mouth, the greater the R value.

6 is a skin lifting measurement photograph of the skin lifting improvement evaluation test of the cosmetic composition.

Table 11 shows the skin lifting improvement rate (%).

Table 11

*; p<0.05

7 shows the measurement results for each week of the skin lifting improvement evaluation test of the cosmetic composition.

Referring to Table 11 and FIG. 7 , it was confirmed that the cosmetic composition according to the present invention helped improve skin lifting at a statistically significant level (p<0.05) after 8 weeks.

In addition, the cosmetic composition according to the present invention is a safe composition that does not irritate the skin when used as a cosmetic, as no symptoms related to skin adverse reactions were reported in skin safety evaluation by a specialist and questionnaire evaluation for subjects. Confirmed.

Claims (5)

A novel palmitoyl tripeptide compound represented by the following structural formula I or palmitoyl-glycinyl-prolinyl-hydroxyproline (Palmitoyl-Glycinyl-Prolinyl-Hydroxyproline; Palm-Gly-Pro-Hyp).
[Structural Formula I]

Using 2-(4-nitrophenyl)sulfonylethoxycarbonyl-amino acid (Nsc-amino acid) as a reactant, glycinyl-prolinyl-hydroxypropyl was obtained by sequentially bonding amino acids starting from a resin three times. synthesizing rollinyl-resin (Gly-Pro-Hyp-resin);
Palmitoyl-glycinyl-prolinyl-hydroxyproline-resin by introducing a palmitoyl group into the glycinyl-prolinyl-hydroxyprolinyl-resin (Palm-Gly-Pro-Hyp-resin) synthesizing; and
Comprising the step of removing the resin and the side chain protecting group from the palmitoyl-glycinyl-prolinyl-hydroxyprolinyl-resin,
Method for preparing the palmitoyl tripeptide compound of claim 1 (Palm-Gly-Pro-Hyp) A cosmetic composition for wrinkle improvement and anti-aging containing the palmitoyl tripeptide compound of claim 1. 4. The method of claim 3,
The cosmetic composition is a cosmetic composition, characterized in that one selected from the group consisting of a flexible lotion (skin), a nourishing lotion, and a nourishing cream. 5. The method of claim 3 or 4,
The content of the palmitoyl tripeptide compound is a cosmetic composition, characterized in that 0.0001 to 0.1% by weight.

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