KR100721144B1 - Cosmetic composition for caring wrinkle containing peptide combination - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin wrinkles, the tripeptide of SEQ ID NO: 1 having a palmitoyl group bonded to the N-terminus, a tetrapeptide of SEQ ID NO: 2 having a palmitoyl group bonded to the N-terminus, N- Provided is a cosmetic composition comprising a peptide mixture consisting of a tetrapeptide of SEQ ID NO: 3 having an acetyl group bonded to its end and a pentapeptide of SEQ ID NO: 4 having a palmitoyl group bonded to its N-terminus. The cosmetic composition according to the present invention exhibits a synergistic effect in improving skin wrinkles by mixing four peptides of different amino acid number and arrangement, and improves the effect of improving skin wrinkles, safety and storage stability by preparing such peptide mixtures in liposome form. Can be.

Peptide Mixture, Improves Wrinkles, Synergies, Liposomes

Description

Cosmetic composition for improving wrinkles comprising a peptide mixture {Cosmetic composition for caring wrinkle containing peptide combination}

1 is a particle size analysis result of the peptide mixture liposome which is an embodiment according to the present invention.

2 is a Zeta potential measurement result of the peptide mixture liposomes according to an embodiment of the present invention.

The present invention relates to a cosmetic composition for improving wrinkles containing peptide excellent in skin safety, storage stability and skin wrinkle improvement.

Skin, which is composed of epidermis, dermis and subcutaneous fat, is the largest organ that makes up the human body, and is a very important organ that plays various roles such as temperature control function, excretory function, respiratory function, human protection function, and barrier function. However, as the skin ages, the function of the skin decreases drastically, and as a result, various changes occur and aging progresses. In particular, active oxygen and free radicals generated due to increased UV and air pollution, excessive fatigue and stress of modern people are known to act as a major cause of skin aging by oxidizing or degenerating biological components due to their high reactivity. To date, no clear cause has been identified.

The physiological changes of the skin by aging include the thinning of epidermis, dermis, and subcutaneous tissue, and the lipid composition and content of the lipid barrier, which functions as a skin barrier, and its function rapidly increases. There is a variety of changes, such as lowering the moisture content of the skin, the skin is dried, blemishes, freckles and pigmentation.

Recently, various studies on the change of collagen, which is a major component of ECM and extracellular matrix (ECM), which are closely related to wrinkles and elasticity of the skin, are being conducted. Collagen accounts for 70-80% of the ECM component, which is known to cause wrinkles as its age decreases rapidly. In addition, functional degradation due to oxidation of elastin, proteoglycan, glucosaminoglycan, laminin, fibronectin, etc., which form skin connective tissues in addition to collagen, has been known as a major cause of skin elasticity and wrinkle formation.

The connective tissue fibers of ECM include glue fibers (collagen fibers, collagen fibers), reticular fibers and elastic fibers (elastic fibers, elastic fibers). Collagen, which accounts for about 70% of skin connective tissue, Is formed in most fibroblasts. As we age, collagen content in the skin bonds decreases due to a decrease in collagen synthesis and accelerated degradation. The biosynthesis process of collagen is regulated by many factors involved in transcription level and post-transcriptional level, and collagen degradation is induced by matrix metalloprotease (MMP) such as collagenase, which breaks down collagen by UV light. This is achieved by promoting collagen degradation by promoting expression. In addition to promoting such degradation and degradation, the deformation of collagen is accelerated by the external environment, and the skin becomes wrinkled and deepened. As a result, skin aging phenomenon such as sagging skin, loss of elasticity, and wrinkled skin is caused by inhomogeneity of cells, loss of elastin, destruction of collagen, decrease and delay in collagen synthesis. Thus, skin aging occurs in the skin epidermis, but rather in the dermis.

Various cosmetic compositions have been researched to solve the problems of skin aging, and the wrinkle improvement effect of the skin has shown visible results in some. There are various reports of clinical results of skin wrinkle removal for retinoids, especially retinol (vitamin A), which are widely used to improve skin wrinkles, blemishes, and pigmentation. Skin sagging, elasticity reduction, etc. are effectively improved, US Patent Nos. 4,603,146, 4,877,805, etc. There are cases of wrinkle improvement using retinol effective for collagen synthesis and degradation inhibition. In addition, cosmetic compositions containing L-ascorbic acid (vitamin C), which are essential for the collagen synthesis process, have been reported to be effective for skin wrinkle improvement, sun protection, pigmentation such as blemishes, freckles, and black mushrooms (US Patent No. 4,938,969). No. 4,772,591 and EP 533,667).

However, the above-described skin improvement active ingredients have many problems in safety and stability to be used as a raw material of cosmetics, and cases of solving them have also been reported. Retinol, which promotes collagen synthesis and improves skin wrinkles, has been difficult to oxidize in light, moisture, air, heat, etc., and is very difficult to be used as a raw material for cosmetics.However, US Pat. No. 4,720,353, BHA (butylated hydroxy anisole) as an antioxidant, vitamin Stabilized using C, propyl gallate and tocopherol linoleate, European Patent No. 440,398 discloses water-in-oil containing one or more water soluble antioxidants, one or more useful antioxidants, and chelates (W / O). The retinol was stabilized with an emulsion type). In addition, various liposomes and capsule formulations for stabilizing retinol have been studied and used in cosmetics, but even if the stability aspect as a cosmetic problem, skin irritation problems still remain in use.

Therefore, the technical problem to be achieved by the present invention is to provide a cosmetic composition that is safe in use, has a high storage stability, and has an excellent wrinkle improvement effect.

In order to achieve the above technical problem, the present invention is a tripeptide (Palmitoyl-Glycyl-Histidyl-Lysine) of SEQ ID NO: 1 is a palmitoyl group bonded to the N-terminal, SEQ ID NO: : Tetrapeptide (Palmitoyl-Glycyl-Glutaminyl-Prolyl-Arginine) of 2, tetrapeptide (Acetyl-Lysine-Aspartic Acid-Valine-Tyrosine) having the acetyl group bonded to the N-terminus and Palmi at the N-terminus It provides a cosmetic composition for improving skin wrinkles, comprising a peptide mixture consisting of pentapeptide (Palmitoyl-Lysyl-Threonyl-Threonyl-Lysyl-Serine) having a toyl group bound thereto.

More preferably, the present invention provides a cosmetic composition for improving skin wrinkles, characterized in that the peptide mixture is prepared in the form of liposomes.

The present inventors used a mixture of peptides having different functions with different amino acid numbers while studying a cosmetic composition having an excellent skin wrinkle improvement effect. As a result, the synergy effect not only increases the skin wrinkle improvement effect. It is confirmed that the skin is safe, and based on this, the present invention was completed.

Hereinafter, the cosmetic composition for improving skin wrinkles containing the peptide mixture of the present invention will be described in more detail.

The cosmetic composition of the present invention comprises a tripeptide of SEQ ID NO: 1 having a palmitoyl group bonded to the N-terminus. The palmitoyl tripeptide is a biosimilar protein, which is relatively smaller in molecular weight than other peptides, and thus easily penetrates the epidermal layer and has excellent skin absorption in the dermal layer. This effectively works on fibroblasts in the dermal layer, effectively enhancing collagen synthesis and synthesis of glycosaminoglycans, which not only improves skin moisturization but also inhibits wrinkle formation of the skin.

The cosmetic composition of the present invention also comprises a tetrapeptide of SEQ ID NO: 2 having a palmitoyl group bonded to the N-terminus. The palmitoyl tetrapeptide has a function similar to that of DHEA (dehydroepiandrosterone), which inhibits cytokine production such as IL6 in keratinocytes and fibroblasts, and has an excellent effect on delaying aging and promotes the formation of keratinocytes and fibroblasts. The fine wrinkle effect on the skin is excellent.

The cosmetic composition of the present invention also comprises a tetrapeptide of SEQ ID NO: 3 having an acetyl group bonded to the N-terminus. These acetyltetrapeptides are bio-like tetrapeptides reconstituted only with their active sites to have a stronger activity of a polypeptide specifically present only in young people. Acetyl tetrapeptides increase skin immunity and regenerate components of the skin by compensating for the loss of thymus factors associated with the degeneration of the thymus with aging. This component functions to promote the production of GM-CSF by keratinocytes. This cytokine promotes the growth of immune cells, such as Langerhans cells, which improves the skin's immune defenses. In addition, it promotes the synthesis of fibronectin, laminin, elastin and collagen to strengthen dermal cell tissues, thereby improving skin elasticity and wrinkle improvement.

The cosmetic composition of the present invention also includes a pentapeptide of SEQ ID NO: 4 having a palmitoyl group bonded to the N-terminus. Such palmitoyl pentapeptides can easily penetrate the epidermal layer of the skin by palmitoyl group and easily penetrate the dermal layer by the hydrophilic amino acid group to effectively act on fibroblasts present in the dermal layer. Therefore, even at very low concentrations (10 ^-9 -10 ^-12 M), it promotes the synthesis of fibronectin and β-TGF, a growth factor that facilitates the intercellular communication of information about collagen synthesis and cell adhesion. . In addition, as a major component of the extracellular matrix of the dermis, it effectively promotes the synthesis of glycosaminoglycans, which play an important role in moisturizing the skin.

The present invention is based on the surprising finding that cosmetic compositions prepared using peptide mixtures comprising all four polypeptides have about 20 to 40% more wrinkle improvement than cosmetic compositions comprising each polypeptide alone. .

Preferably, the present invention provides a cosmetic composition for improving skin wrinkles, characterized in that the peptide mixture is prepared in a liposome form. The peptide mixture may be encapsulated in liposomes to effectively penetrate the skin, thereby maximizing the effect of the peptides on the skin, and improving storage stability. Such liposomes are well known to those skilled in the art to which the present invention pertains, and can be prepared using, for example, unsaturated lecithin, saturated lecithin, but are not limited thereto. Liposomes formed based on such unsaturated lecithin and saturated lecithin are not only excellent in affinity for the skin but also are manufactured in a very uniform and fine size, so that the penetration into the skin can be greatly improved.

The present invention provides a cosmetic composition containing 0.1 to 90% by weight of the peptide mixture-containing liposomes relative to the total weight of the cosmetic composition. Also preferably, the present invention is the palmitoyl tripeptide, palmitoyl tetrapeptide, acetyl tetrapeptide and palmitoylpentapeptide are 5-30% by weight, 1-20% by weight, and 1-10, respectively, relative to the total weight of liposomes containing the peptide mixture. It provides a cosmetic composition for improving skin wrinkles, characterized in that it contains by weight and 1-20% by weight, more preferably, the present invention is palmitoyl tripeptide, palmitoyl tetrapeptide, acetyl tetrapeptide and palmitoyl penta peptide It provides a cosmetic composition for improving skin wrinkles, characterized in that it comprises 10-15% by weight, 3-7% by weight, 1.5-3.5% by weight and 3-7% by weight relative to the total weight of the liposome-containing peptide mixture. Cosmetic composition according to the present invention having such a composition ratio is excellent in the improvement effect of the skin wrinkles, excellent formulation stability and can be used safely without side effects on the skin.

The formulation of the cosmetic composition to be applied is not particularly limited, for example, supple cosmetics, astringent cosmetics, nourishing cosmetics, eye cream, nutrition cream, massage cream, cleansing cream, cleansing foam, cleansing water, powder, essence, pack, etc. It may be prepared in a dosage form.

Hereinafter, the present invention will be described in detail with reference to Examples. However, embodiments according to the present invention may be modified in many different forms, and the scope of the present invention should not be construed as being limited to the embodiments described below. The embodiments of the present invention are provided to more completely explain the present invention to those having ordinary skill in the art.

Preparation of Peptide Mixture Liposomes

Liposome preparation containing the peptide of the present invention and the preparation was prepared in the composition and content shown in Table 1 below. Unsaturated lecithin, saturated lecithin, ethanol, oleic acid, cholesterol, and palmitic acid are heated to about 70 ° C to completely dissolve, then mixed with the remaining dissolved aqueous phase to give the mixture a 20 minute wet time. Was stirred. The homogenized mixture was passed three times at 50 ° C. and 1000 bar with a microfluidizer to prepare a peptide mixture in liposome form.

In Table 1, the unit is% by weight, and the tripeptide is palmitoyl-glycile-histidyl-lysine (Biopeptide CL ™ (100 ppm), Sederma, France), and the tetrapeptide ① is palmitoyl-glysil-glutami. Nyl-prolyl-arginine (RISIN ™ (500 ppm), Sederma, France), tetrapeptide ② is acetyl-lysine-aspartic acid-valine-tyrosine (Thymulen 4 ™ (100 ppm), Atrium, Canada), pentapeptide Palmitoyl-lysyl-threonyl-threonyl-lysyl-serine (Matrxyl ™ (100 ppm), Sederma, France).

<Measurement of particle size of peptide mixture liposomes>

The size of liposomes was measured using a light scattering device, Malvern system (Malvern PCS 4700, Malvern Instruments, UK). Malvern systems feature He-Ne lasers (Siemens, LGK 7626), PMT (PCS100), step-moto regulators (PCS7), correlators (K7032ES), pump / filter units (PR98) and thermostats with a wavelength of 633 nm. (PCS8) and light scattering data were analyzed with Malvern 4700 software. Temperature control in the vat was maintained constant at 25 ± 0.01 ℃ using PCS8. Dust removal in the vat was considered clean if the scattering intensity value measured by PMT was less than 0.2 Kcps. The average value of each sample was measured 10 times at 90 ° and the average value is shown in Table 2 below. 1 shows a particle size analysis picture of the preparation example.

Zeta potential measurement of peptide mixture liposomes

Zeta potentiometer (Malvern, Zetasizer 2000, Malvern Instrument, UK) was used to examine the stability of the particles.The DTS0050 standard electrolyte was measured to confirm the value within the range of ± 5㎷. It was. The average value was measured five times for one sample and the average value is shown in Table 2. 2 shows the Zeta potential measurement results of Preparation.

As shown in Table 2, the average particle size and Zeta potential of the peptide mixture liposome particles prepared based on unsaturated and saturated lecithin were 77.8 to 86.2 nm and -48.3 to -55.8 각각, respectively. It can be seen that liposomes have been prepared.

<Production of Examples and Comparative Examples>

To prepare a cosmetic composition comprising a peptide as shown in Table 3. Comparative Examples 1 to 7 are cosmetic compositions in which the constituents of the peptide as an active ingredient are added with different peptides or peptide mixtures, and Comparative Examples 8 and 9 do not contain peptides and are cosmetics containing retinol and vitamin C which are different active ingredients, respectively. Composition.

In Table 3, the unit is weight percent, Retinol was used as Retinol Wax ™ (Mibelle AG, Switzerland), and Vitamin C was used as Vitamin C ™ (ROCHE, France).

The following test was done about the cosmetic composition of the said Example and a comparative example.

<Measurement of collagen synthesis promotion effect ( in vitro )>

Human normal fibroblasts were seeded in 48-well microplates containing DMEM medium containing 10% fetal calf serum (1 × 10 ⁵ cells / well) and incubated at 37 ° C. for 24 hours in a 5% CO ₂ incubator. It was. Normal human fibroblasts were isolated from skin tissues collected after circumcision of newborn in hospital. Samples of Examples and Comparative Examples were added to DMEM medium without serum to a final concentration of 20 μg / ml, and then cultured for 24 hours after replacement with the medium, followed by cell culture. The collagen content of the collected cell culture solution was measured using a collagen protein measurement kit (Takara Shuzo Co., Ltd. Japan). First, the cell culture fluid was collected into 96-well microplates uniformly coated with primary collagen antibodies, and antigen-antibody reactions were caused at 37 ° C. for 3 hours. After the reaction, the reaction solution in the well was removed and washed four times with PBS buffer. Subsequently, the color-causing substance was added to each well and reacted at room temperature for 15 minutes. 1 N sulfuric acid solution was added to each well containing the reaction solution to stop the reaction, and the absorbance at 450 nm was measured with a spectrophotometer.

Standard concentrations were obtained by substituting respective concentrations into absorbances of the standard solutions, and the amount of collagen produced was calculated by substituting the reaction absorbances of the respective cultures with the extracts into the standard curves. The collagen synthesis promotion rate was calculated according to the following Equation 1, and the results are shown in Table 4 below.

Collagen synthesis promotion rate (%) = (A × 100) / B-100

In Equation 1, A is the collagen production amount of the active ingredient-added culture, B is the collagen production amount of the culture medium without the sample.

As shown in Table 4, the embodiment prepared by mixing the four peptides of 17.1-35.2% compared to Comparative Examples 2 to 7 using one kind of peptide alone or other peptide mixtures other than the mixed composition according to the present invention. It can be seen that the collagen synthesis promotion rate is high. It can be seen that there is a synergistic effect on promoting collagen synthesis when using the peptide mixture as described above.

<Measurement of fibroblast synthesis efficacy ( in vitro )>

10,000 cells were inoculated in 96-well and incubated for 12 hours, and the samples were incubated for 1, 2, 3 and 4 days, followed by MTT (3- [4,5-dimethylthiaxol-2-yl] -2. 5 mg / ml of 5-diphenyltetrazolium bromide) was added to the wells and incubated for 4 hours. Thereafter, the medium was removed, 100 μl of DMSO was added thereto, the mixture was stirred for 5 minutes, and the absorbance was measured at 570 nm with an ELISA reader. The results are shown in Table 5 below.

As shown in Table 5, the cosmetic composition comprising the peptide mixture according to the present invention is 12.7-35.2 compared to Comparative Examples 2 to 7 using only one type of peptide alone or other peptide mixtures other than the mixed composition according to the present invention. It can be seen that the percentage of fibroblast synthesis promoting rate is high. It can be seen that there is a synergistic effect on promoting fibroblast synthesis when using the peptide mixture as described above.

Clinical Effect of Skin Wrinkle Improvement

Thirty women aged 30 years or older (mean age 34.3 years) were divided into three groups (A, B, and C), the sample of Example in group A, the sample of Comparative Example 5 in group B, and the sample of Comparative Example 6 in group C. After the sample was applied for 12 weeks (2 times / day) around each wrinkled eye area, the subject's subjective evaluation and subject's subjective evaluation were classified into the following six levels to evaluate the improvement of skin wrinkles (△ W) was measured. The results are shown in Tables 6 and 7, respectively.

-Skin wrinkle improvement evaluation criteria-

-3: extremely badly deteriorated

-2: worse

-1: slightly worse

0: no change

1: slightly improved

2: improve

3: highly improved

As shown in Tables 6 and 7, the embodiment using the peptide mixture according to the present invention can be seen that the objective evaluation of the expert and the subjective evaluation of the subject is 2.6 and 2.7, respectively, the wrinkle improvement effect is very excellent compared to the comparative examples have. Through this, it can be seen that the wrinkle improvement effect is very effectively increased when using the peptide mixture as in the present invention.

<Formulation stability test>

Examples and Comparative Examples 1-9 were stored at 4 ° C., room temperature (25 ° C.), cycling (-4 ° C. to 45 ° C., twice a day) and 45 ° C. for 4 weeks, and then the degree of separation and discoloration were measured. Evaluation was made using the naked eye and a colormeter, respectively. The criterion of comparison was 0 for no color change and 5 for severe case. The experimental results are shown in Table 8 below.

Evaluation criteria for visual separation are as follows.

○ = good, △ = start of separation, × = complete separation

As shown in Table 8, in Comparative Examples 8 and 9 showed a severe discoloration and formulation stability at high temperatures and cycles, while in the Example it was confirmed that there is almost no degree of separation and discoloration at high temperature and room temperature .

<Safety test>

Patch tests were conducted to test the safety of the samples of Examples and Comparative Examples 1-9. Twenty subjects (M: 6 / F: 14) who participated in the test were treated with the appropriate concentrations of each test substance in the Finn chamber and affixed to the inner region of the upper arm. Observed and evaluated, and the number of subjects corresponding to each degree of stimulation is shown in Table 9.

As shown in Table 9, Examples and Comparative Examples 1-7 including the peptide according to the present invention can be seen that the safety for the skin is excellent. However, in Comparative Examples 8 and 9 containing retinol and vitamin C, erythema, swelling and irritation were observed to be maintained for a considerable time.

The present invention is a tripeptide of SEQ ID NO: 1 having a palmitoyl group bonded to the N-terminus, a tetrapeptide of SEQ ID NO: 2 having a palmitoyl group bonded to the N-terminal sequence, and an acetyl group having a acetyl group bound to the N-terminal: Provides a cosmetic composition for improving skin wrinkles comprising a peptide mixture consisting of a tetrapeptide of 3 and a pentapeptide of SEQ ID NO: 4 having a palmitoyl group bonded to the N-terminus. The cosmetic composition according to the present invention exhibits synergistic effect in improving skin wrinkles by mixing four peptides of different amino acid number and arrangement, and can improve skin wrinkle improvement and safety by stabilizing the peptide mixture in liposome form. .

Attach sequence list electronic file

Claims (6)

A tripeptide of Palmidyl number 1 (Palmitoyl-Glycyl-Histidyl-Lysine) having a palmitoyl group bonded to an N-terminus, and a tetrapeptide (Palmitoyl-Glycyl-Glutaminyl) of SEQ ID NO: 2 having a palmitoyl group bound to an N-terminal -Prolyl-Arginine), a tetrapeptide of SEQ ID NO: 3 having an acetyl group bound to the N-terminus (Acetyl-Lysine-Aspartic Acid-Valine-Tyrosine), and a penta of SEQ ID NO: 4 having a palmitoyl group bound to the N-terminus Peptides (Palmitoyl-Lysyl-Threonyl-Threonyl-Lysyl-Serine) Cosmetic composition for improving skin wrinkles comprising a peptide mixture consisting of. The cosmetic composition for improving skin wrinkles according to claim 1, wherein the peptide mixture is prepared in the form of liposomes. The cosmetic composition for improving skin wrinkles according to claim 2, wherein the liposome is prepared using an unsaturated lecithin and a saturated lecithin. The cosmetic composition for improving skin wrinkles according to claim 2, wherein the peptide mixture-containing liposome is contained in an amount of 0.1 to 90% by weight based on the total weight of the cosmetic composition. According to claim 2, wherein the tripeptide of SEQ ID NO: 1 is a palmitoyl group bonded to the N-terminal, the tetrapeptide of SEQ ID NO: 2 is a palmitoyl group bonded to the N-terminal, acetyl group is bonded to the N-terminal The tetrapeptide of SEQ ID NO: 3 and the pentapeptide of SEQ ID NO: 4 having a palmitoyl group bonded to the N-terminus are 5-30% by weight, 1-20% by weight, and 1-10% by weight of the total weight of liposomes containing the peptide mixture. Cosmetic composition for improving skin wrinkles, characterized in that contained by weight and 1-20% by weight. According to claim 5, wherein the tripeptide of SEQ ID NO: 1 is a palmitoyl group bonded to the N-terminal, the tetrapeptide of SEQ ID NO: 2 is a palmitoyl group bonded to the N-terminal, acetyl group is bonded to the N-terminal The tetrapeptide of SEQ ID NO: 3 and the pentapeptide of SEQ ID NO: 4 having a palmitoyl group bonded to the N-terminus are 10-15%, 3-7%, and 1.5-3.5, respectively, based on the total weight of liposomes containing the peptide mixture. Cosmetic composition for improving skin wrinkles, characterized in that contained by weight and 3-7% by weight.

Images