KR20230078893A - Preparing method of peptide with improved heat and enzymatic hydrolysis stability and cosmetic composition using the same - Google Patents

Abstract

An improved peptide that has improved thermal and enzymatic decomposition stability and can be used as a cosmetic composition for manufacturing functional cosmetics is disclosed. The present invention is to provide the improved peptide in which a peptide consisting of an amino acid sequence of Glycine (Gly)-Glutamine-Valine (Val)-Serine (Ser) and a saturated or unsaturated fatty acid of 10 to 20 carbon atoms are amide-linked.

Description

Preparation method of peptide with improved heat and enzymatic hydrolysis stability and cosmetic composition using the same}

The present invention relates to a method for preparing a peptide and a cosmetic composition using the same, and more particularly, to a method for preparing a peptide with improved thermal and enzymatic degradation stability and a cosmetic composition using the same.

The skin is the largest tissue in the human body and functions to protect the inside of the body from sunlight, physical and chemical stimuli, and is absolutely necessary for human life maintenance and is constantly regenerated to maintain homeostasis. The skin is composed of epidermis, dermis, and subcutaneous fat in order from the outside. there is. Skin is a tissue that covers the entire body and has various functions. Since it has a barrier function between the inside and outside of the body, it has an immune system by the islets of Langerhans and has a moisture control function by the epidermis. In addition, it synthesizes vitamin D with appropriate sunlight and performs the function of lipid storage and excretion through sweat glands. The stratum corneum produced by the normal differentiation process in healthy skin has the function of maintaining moisture in the skin and protecting it from external environmental stimuli. This function is called the skin barrier function and can be said to be the most important role of the epidermis.

One of the areas of greatest interest in cosmetics is aging related to aging, pigmentation, loss of elasticity, skin dryness, hair loss, and lack of luster in hair. The skin undergoes various changes through aging. First, the thickness of the epidermis, dermis, and subcutaneous tissue, which are components of the skin, becomes thinner, and the extracellular matrix (ECM) component that gives skin elasticity changes. As collagen production rapidly decreases with age, it has a close relationship with aging generation, and collagen, elastin, proteoglycans, and glucosaminoglycans ( Glucosaminoglycan), laminin, fibronectin, etc. are oxidized and lose their functions, so the skin loses its elasticity and aging is excessively formed, resulting in aging skin.

Connective tissue fibers of the extracellular matrix include glial fibers (collagen fibers), reticular fibers, and elastic fibers (elastic fibers), of which collagen accounts for about 70% of skin connective tissue. is mostly formed in fibroblasts of the skin. With age, the collagen content in skin connective tissue is reduced, which is due to the decrease in collagen synthesis and the promotion of decomposition. Therefore, the decrease in collagen biosynthesis and the acceleration of collagen degradation can be said to be the biggest causes of skin aging.

The process of collagen biosynthesis is regulated by many factors involved in the transcription level and the post-translation level, resulting in changes, and collagen degradation is caused by collagenase, which degrades collagen by ultraviolet rays, etc. By promoting the expression of matrix metalloproteases (MMPs) such as collagenase, collagen degradation is promoted, and the collagen content is reduced. In addition, as the transformation of collagen is accelerated by the external environment, the aging of the skin increases and deepens. As a result, the skin aging phenomenon appears due to non-homogenization of cells, loss of elastin, destruction of collagen, decrease and delay of collagen synthesis, and the like. Therefore, although skin aging occurs in the epidermis, it can be seen as a phenomenon that occurs in the dermis rather than in the epidermis.

Various cosmetic compositions are being studied to solve the problem of skin aging, and some of the skin aging reduction effects are showing visible results. For example, retinoids widely used to improve skin aging, melasma, and pigmentation, and in particular, clinical results for removing skin aging for retinol have been reported in various ways, and cosmetics containing retinol can reduce skin aging caused by sunlight. Effectively improved weight, skin sagging, and loss of elasticity.

In recent years, as human lifespan has increased and the standard of living has improved, people's interest in becoming healthy and beautiful has increased. It can be well known that

Skin aging has been divided into extrinsic aging (aging caused by external stimuli such as sunlight) and intrinsic aging (natural aging due to age). Aged skin has increased wrinkles and decreased elasticity. However, understanding of the causes of these skin aging symptoms, prevention and treatment techniques have not yet been developed. As a result, skin aging occurs due to non-homogenization of cells, loss of elastin, destruction of collagen, reduction and delay of collagen synthesis, etc. Therefore, skin aging occurs in the skin epidermis, but rather in the dermis.

Various cosmetic compositions are being studied to solve the problem of skin aging, and among them, peptides are biomolecules produced through the polymerization of amino acids, which are harmless to the human body and are known as substances that regulate various physiological activities in vivo. . In particular, peptides are drugs widely used in medicine and pharmacies because they are very stable against heat and pH and do not easily lose their activity compared to proteins, which are large combinations of amino acids. . However, various attempts have been made to develop various anti-wrinkle agents using peptides, but the effect was not so great, and vitamin C-modified peptides and hydroquinone-modified peptides to maximize the effect are also difficult to manufacture and expensive. Because of this, its use is limited, so there are not many applications in the market. In particular, since peptides are substances with good solubility in water and insoluble in oil, they have physical properties that are difficult to penetrate into the skin through the skin layer and exhibit physiologically active functions.

In order to smoothly synthesize a peptide, a protecting group is required. Until now, protecting groups with various characteristics have been developed and used. As a representative protecting group, Fmoc/trt-amino acid derivatives have been developed and reported to have excellent effects when used in some peptide synthesis. However, even though Fmoc/trt-amino acid derivatives show excellent effects in solid-phase peptide synthesis, Nevertheless, low condensation yield was shown in the synthesis of egg-structured peptides due to low solubility due to hydrophobicity of some Fmoc. On the other hand, 2-(4-nitrophenyl)sulfonylethoxycarbonyl (Nsc), due to the structural characteristics of the nitro group and the sulfonyl group, increases the solubility of amino acids and suppresses the hydrophobic interaction between peptide chains. play a role In addition, Nsc has excellent stability compared to Fmoc under an automated system, and thus has a very advantageous advantage in synthesizing long-chain peptides.

On the other hand, peptides with many advantages are generally stable to heat and pH, but have the disadvantage of being weak against high temperatures of 60 ° C or higher and proteases. In the field of general medicines or cosmetics, it is rarely stored or distributed at high temperatures, but in some manufacturing processes, high temperature conditions of 80 ° C or higher are required, or peptides are decomposed due to various proteolytic enzymes outside and inside the skin.

The present invention aims to provide an improved peptide with improved stability through binding with various substituents to the highly functional peptide filed by the present applicant on November 25, 2020 to overcome the above-mentioned disadvantages and a cosmetic composition using the same .

As a first aspect to solve the above problem, the present invention provides a peptide consisting of an amino acid sequence of glycine (Gly)-glutamin-valine (Val)-serine (Serine, Ser) and carbon atoms of 10 to 20 Provides an improved peptide in which saturated or unsaturated fatty acids of amide are bonded.

In addition, it provides an improved peptide, characterized in that the fatty acid is palmitic acid.

In addition, as a first aspect to solve the above problem, the present invention, glycine (Glycine, Gly) - glutamine (Glutamin) - valine (Valine, Val) - serine (Serine, Ser) peptide consisting of an amino acid sequence and an acetyl group amide Conjugated improved peptides are provided.

In addition, the improved peptide is synthesized using 2-(4-nitrophenyl)sulfonylethoxycarbonyl-amino acid (Nsc-amino acid) as an intermediate.

In addition, the improved peptide provides an improved peptide characterized in that the thermal stability is improved compared to the peptide.

In addition, the improved peptide provides an improved peptide characterized in that protease resistance is improved compared to the peptide.

In order to solve the above another problem, the present invention provides a cosmetic composition containing the improved peptide.

The improved peptide according to the present invention has improved thermal and enzymatic decomposition stability, so it can be used as a cosmetic composition for producing functional cosmetics.

1 is a view showing the peptide synthesized in Example 1 of the present invention.
Figure 2 is a graph showing the MS (Mass spectrometry) analysis results of the peptide in Example 2 of the present invention.
Figure 3 is a graph showing the thermal stability test results of the peptide in Example 3 of the present invention.
Figure 4 is a graph showing the test results of the degradation enzyme stability of the peptide in Example 4 of the present invention.
Figure 5 is a graph showing the results of the cytotoxicity test of the peptide in Example 5 of the present invention.

Hereinafter, the present invention will be described in detail through preferred embodiments. Prior to this, the terms or words used in this specification and claims should not be construed as being limited to the usual or dictionary meaning, and the inventor appropriately uses the concept of the term in order to explain his/her invention in the best way. Based on the principle that it can be defined, it should be interpreted as meaning and concept consistent with the technical spirit of the present invention. Therefore, since the configurations of the embodiments described in this specification are merely the most preferred embodiments of the present invention and do not represent all of the technical spirit of the present invention, various equivalents and modifications that can replace them at the time of the present application It should be understood that there may be Also, throughout the specification, when a part "includes" a certain component, it means that it may further include other components, not excluding other components, unless otherwise stated.

The present invention is an improved peptide in which a peptide consisting of an amino acid sequence of glycine (Gly)-glutamin-valine (Val)-serine (Ser) and a saturated or unsaturated fatty acid having 10 to 20 carbon atoms are amide-bonded. And, a peptide consisting of an amino acid sequence of Glycine (Gly)-Glutamine-Valine-Valine-Serine (Ser) and an improved peptide in which an acetyl group is amide-bonded.

In addition, the present invention discloses a cosmetic composition containing the improved peptide.

The improved peptide may be contained in an amount of 0.001 to 10% by weight, preferably 0.01 to 5% by weight, more preferably 0.05 to 1% by weight, based on the total cosmetic composition.

The cosmetic composition according to the present invention may be applied in a gel type, skin type, cream type, ointment type, etc., but is not limited thereto. The above composition can be appropriately prepared by a known method by adding suitable conventional softeners, emulsifiers, thickeners or other materials known in the art according to its type.

The gel-type composition may be prepared by adding a softening agent such as trimethylolpropane, polyethylene glycol or glycerin, a solvent such as propylene glycol, ethanol, or isocetic alcohol, purified alcohol, or the like.

The skin type composition contains fatty alcohols such as stearyl alcohol, myristyl alcohol, behenyl alcohol, arachidyl alcohol, isostearyl alcohol and isocetyl alcohol, butylene glycol, glycerin, allantoin, methyl paraben, EDTA- It can be prepared by adding 2-sodium, xanthan gum, dimethicone, polyethylene glycol-60 hydrogenate, castol oil, polysorbate 60, and purified water.

The cream-type composition includes fatty alcohols such as stearyl alcohol, myristyl alcohol, behenyl alcohol, arachidyl alcohol, isostearyl alcohol, and isocetyl alcohol, lecithin, phosphatidylcholine, phosphatidylethanolamine, sofphatidylserine, sofphatidyl Lipids such as inositol, derivatives thereof, emulsifiers such as glyceryl stearate, sorbitan palmitate, and soribitan stearate, natural fats or oils such as avocado oil, apricot oil, babassu oil, vitreous borage oil, camellia oil, It can be prepared by adding a solvent such as propylene glycol and purified water.

The ointment-type composition may be prepared by adding a softener, an emulsifier, and a wax such as microcrystalline wax, paraffin, ceresin, beeswax, spermaceti, and vaseline.

The cosmetic composition of the present invention may further contain at least one skin wrinkle improvement component exhibiting the same or similar function in addition to the active ingredient. The skin wrinkle improvement component may be at least one selected from the group consisting of vitamin C, retinoic acid, TGF, animal placenta-derived protein, betulinic acid, and chlorella extract, but is not limited thereto. In addition, the cosmetic composition of the present invention may further contain excipients including fluorescent substances, fungicides, hydrotropes inducing substances, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, plant extracts, and the like. can

In the present invention, the cosmetic composition is softening lotion, astringent lotion, nutrient lotion, eye cream, serum, nutrient cream, massage cream, cleansing cream, cleansing lotion, cleansing foam, cleansing water, powder, essence, pack, hair tonic, hair treatment It can be formulated as a ment, shampoo or rinse.

The cosmetic composition may be formulated by conventional methods. In the formulation of external skin preparations, reference may be made to the contents disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA, and in the formulation of cosmetic compositions, International Cosmetic Ingredient Dictionary, 6th ed (The cosmetic, Toiletry and Fragrance Association , Inc, Washington, 1995) may be referred to.

Specifically, the cosmetic composition may be prepared in general emulsified and solubilized formulations. For example, cosmetic lotion, such as softening lotion or nourishing lotion; emulsions such as facial lotion and body lotion; creams such as nourishing creams, moisture creams, and eye creams; essence; cosmetic ointment; spray; gel; pack; sunscreen; makeup base; foundations such as liquid type, solid type or spray type; powder; makeup removers such as cleansing creams, cleansing lotions, and cleansing oils; Alternatively, it may be formulated as a cleanser such as a cleansing foam, soap, body wash, etc., but is not limited thereto. In addition, the skin external preparation may be formulated as an ointment, patch, gel, cream or spray, but is not limited thereto.

In addition to the essential components in each formulation, the cosmetic composition may be appropriately mixed with other components within a range that does not impair the purpose of the present invention depending on the type of formulation or purpose of use.

The cosmetic composition may include a conventionally acceptable carrier, and for example, oil, water, a surfactant, a moisturizer, a lower alcohol, a thickener, a chelating agent, a colorant, a preservative, a flavoring agent, and the like may be appropriately blended, but are not limited thereto. no.

The acceptable carrier may vary depending on the formulation. For example, when formulated into an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanthate, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or any of these as a carrier component Mixtures may be used.

When the cosmetic composition is formulated as a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or a mixture thereof may be used as a carrier component, and in the case of a spray, chlorofluoro It may further contain a propellant such as hydrocarbon, propane, butane or dimethyl ether.

When the cosmetic composition is formulated as a solution or emulsion, a solvent, solubilizing agent, or emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil may be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, fatty acid esters of glycerol, polyethylene glycol or sorbitan may be used.

When the cosmetic composition is formulated as a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like can be used.

The cosmetic composition includes fatty substances, organic solvents, solubilizers, thickeners, gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, and fragrances commonly used in the industry depending on the quality or function of the final product. , surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives commonly used in cosmetics It may additionally contain adjuvants commonly used in cosmetology or dermatology, such as any other ingredient. However, the adjuvant and its mixing ratio may be appropriately selected so as not to affect desirable properties of the cosmetic composition according to the present invention.

Hereinafter, specific embodiments according to the present invention will be described.

Example 1: Synthesis of improved peptides

19 nsc-amino acids (nsc-Ala, nsc-Arg (pbf), nsc-Asp (OtBu), nsc-Asn (trt), nsc-Gly, nsc-Glu) to construct a peptide library for synthesizing improved peptides (OtBu), nsc-Gln(trt), nsc-His(trt), nsc-Ser(tBu), nsc-Thr(tBu), nsc-Tyr(tBu), nsc-Trp(Boc), nsc-Leu, Chloro trityl chloride resin (CTL resin, Novabiochem Cat No. 01- 64-0021) and 19 Fmoc-amino acids (Fmoc-Ala, Fmoc-Arg (pbf), Fmoc-Asp (OtBu), Fmoc-Asn (trt), Fmoc-Gly, Fmoc-Glu (OtBu), Fmoc- Gln(trt), Fmoc-His(trt), Fmoc-Ser(tBu), Fmoc-Thr(tBu), Fmoc-Tyr(tBu), Fmoc-Trp(Boc), Fmoc-Leu, Fmoc-Ile, Fmoc- Val, Fmoc-Phe, Fmoc-Met, Fmoc-Lys (Boc), Fmoc-Pro) attached to chloro trityl chloride resin (Chloro trityl chloride resin: CTL resin, Novabiochem Cat No. 01-64-0021), Lauroyl chloride, oleoyl chloride, linoleoyl chloride, Myristoyl chloride, Palmitoyl chloride, Stearoyl chloride, a total of 6 fatty acid chlorides, were put into a 96-well Teflon reactor in a 96-well Teflon reactor, and 50 mg was added to each of 19 lines, and 1 ml of methylene chloride (MC) was added to obtain 3 Stir for 1 minute. The solution was removed, 1 ml of dimethylformamide (DMF) was added, stirred for 3 minutes, and then the solvent was removed again. The prepared peptide-resin was washed three times each with DMF, MC, and methanol, dried by slowly flowing nitrogen air, and completely dried under reduced pressure in vacuum under P ₂ O ₅ . To the prepared resin, a fugitive solution (trifluroacetic acid (TFA) 81.5%, distilled water 5%, thioanisole 5%, phenol 5%, EDT (1,2-Ethanedithiol) 2.5% and TIS (Triisopropylsilane 1% included) 30 ml was added, and the reaction was maintained for 1 hour in an ice bath while shaking occasionally at room temperature. The resin was filtered, washed with a small amount of TFA solution and combined with the mother liquor. Afterwards, the peptides shown in FIG. 1 were obtained.

As a result of the above synthesis and purification, glycine (Glycine, Gly)-glutamin-valine (Valine, Val)-serine (Serine, Ser) amino acid sequence (GQVS) peptides, acetyl group bond, palmitate An improved peptide with a palmitoyl group bond and a disulfide bond was obtained (see FIG. 1). The yields of the synthesized peptides and improved peptides differed greatly depending on the three-dimensional structure and physical properties of the peptides, and the yields also showed great differences. It is predicted that the yield during synthesis of the peptide is determined by steric hindrance due to the length of the peptide and the three-dimensional structure of the substituent.

Example 2: MS (Mass spectrometry) analysis of peptides

MS analysis of the peptides and improved peptides in Example 1 was performed using JEOL's JMS-700 High Resolution Mass Spectrometer (MS). At this time, the MS analysis method was measured under the probe bias +4.5 kv condition using the ESI mode, and the MS analysis results are shown in FIG. 2 . Detailed conditions of MS analysis are shown in Table 1 below.

Ion Polarity Positive Nebulizer Gas Flow 1.5 L/min Detector 1.2kv CDL -20.0v CDL Temp 250℃ Block Temp 400℃ T. Flow 0.2ml/min B. conc 50% H ₂ 0 / 50% ACN Probe ESI probe bias +4.5 kv

Referring to FIG. 2 , MS analysis of the peptide and the improved peptide synthesized in Example 1 confirmed that the synthesized peptides were synthesized in the correct sequence through molecular weight. In subsequent examples, the peptide synthesized under the conditions of Example 1 was used.

Example 3: Confirmation of improved thermal stability of modified peptides

In order to confirm the thermal stability of the improved peptide in the present invention, the composition was dissolved in a 50 mM Tris-HCl (pH 8.0) buffer solution so that the concentration of the improved peptide was 10 mg/ml, and then dispensed into glass vials and then incubated at 25 ° C and 25 ° C. It was stored at 70° C. for 1, 2, 4, 8, and 12 weeks, and the stability was measured by measuring the loss of the peptide component in the composition due to heat, and the results are shown in FIG. 3. Stability was determined through HPLC content analysis, and waters' HPLCwaters 2695) and waters' C18 (Waters Xterra MS C18 column (L: 250 mm, LD: 4.6 mm, 5 μm)) columns were used for peptides and improved peptides. and analyzed. At this time, the wavelength of the HPLC detector was 216 nm. Acetonitrile (ACN) containing 0.1% trifluoroacetic acid (TFA) and water containing 0.1% TFA were used as solvents for the mobile phase of HPLC. The purity assay of was observed. Conditions required for analyzing the peptide material are shown in Table 2 below.

Condition of HPLC analysis Column Aminex HPX-87H (300 x 7.8 nm) Detector UVD 170s DIONEX Detection wavelength 216 nm Flow rate 0.7ml/min Injection volume 10 μl Mobile phase
conditions for HPLC
gradient-elution Program order Time (min) 0.1% TFA1)
in DW (%) 0.1% TFAs
in 50% ACN(%) One 0 100 0 2 10 75 25 3 20 50 50 4 30 25 75 5 40 0 100 6 45 100 0

Referring to FIG. 3, as a result of measuring the loss of peptides due to heat, there was an improvement in stability compared to general peptides. In particular, improved peptides in which a palmitoyl group was substituted at the N-terminus had the highest stability improvement It was confirmed that it worked.

Example 5: Degradation enzyme stability confirmation of improved peptide

Experiments for measuring the protease activity of the improved peptides were performed by modifying Kunitz's method. Substrate was used by adding 0.6% casein to 50 mM sodium phosphate buffer, and 0.44 M trichloroacetic acid (Duksan, Korea) was added to the buffer to stop the enzyme reaction. Proteolytic enzyme activity was measured using absorbance change according to the principle of reducing color development. 0.25 ml of the diluted sample solution of each peptide was added to 0.6% casein, and the reaction was carried out at 37℃ for 15 minutes, 30 minutes, 1, 2, 4, and 8 hours, and then 0.44 M trichloroacetic acid was added and the The enzymatic reaction was stopped for a minute. The supernatant obtained by centrifuging the sample solution in which the reaction was stopped was analyzed by HPLC in the same manner as in Example 3 to confirm the stability improvement effect, and the results are shown in FIG. 4 .

Referring to FIG. 4, the obtained improved peptide in which palmitoyl group and acetic acid residue (acetyl group) were substituted at the N-terminus had a remarkable effect on improving stability, and in the case of disulfide-linked peptides, It was confirmed that the resistance was lowered.

Example 5: Cytotoxicity test

In order to confirm whether or not the improved peptide according to the present invention has keratinocyte endotoxicity, cytotoxicity and regenerative ability were evaluated using MTT assay. HaCat cells were seeded in a 24 well plate at a cell count of 4×10 ⁴ cells/24 well per well, and incubated for 24 h at 37° C. under 5% CO ₂ conditions. Thereafter, the cells were washed twice with PBS, cultured, and treated with peptides at different concentrations (50, 500, 1,000 μM) and incubated for 24 h. After incubation, MTT 0.5% in DPBS was mixed and added with the culture medium at a ratio of 1:9 (v/v), cultured in a CO ₂ incubator for 2 h, and the formazan produced was dissolved in DMSO and measured at 570 nm using ELISA. And the results are shown in Figure 5.

Referring to FIG. 5 , with respect to HaCat cells, a type of keratinocyte cell line, no decrease in the number of cells was observed by the composition administered from low to high concentrations, and no significant change was observed in microscopic observation of the cells.

Through the above results, it can be predicted that the improved peptide according to the present invention is not toxic to skin-related cells at all, and thus will not have a significant effect even when the cosmetic composition according to the present invention is treated on the skin.

Preparation Example: Preparation of cream-type composition containing improved peptides

A cream type composition containing the improved peptide was prepared by the following method. As a cream-type composition, 0.1 part by weight of Palmitoyl-GQVS obtained in Example 1 was included, and formulations were configured as shown in Table 3 below.

1) [Prime B] Manufacturing

While warming water to 60 to 70 ° C. while stirring, powdery raw materials are gradually added sequentially, stirred with a disper mixer until thickened, evenly dissolved, then cooled, and prepared in advance for ease of processing.

2) [Phase A] Manufacturing

After sufficiently impregnating the powdery raw material in glycerin, water is added and heated to 50 to 60° C. to dissolve evenly with a disper mixer.

3) [Phase A] + [Phase B] mixing and stirring preparation

Water phase B prepared in advance is added to water phase A and warmed while mixing evenly. Prepare so that 65 to 75 ° C can be maintained.

4) [Oil Phase] Mixing and Stirring Manufacturing

Weigh each raw material in an individual beaker and dissolve it by heating and stirring (at least 80 ° C). The melted phase is prepared so that the temperature can be maintained above 75 ℃.

5) Whole Cream Preparation

The oil phase is gradually added to the prepared water phase mixture, and the emulsification process is performed by vigorously stirring (6,000 rpm) for about 10 minutes. At this time, the temperature is maintained at 65 ~ 75 ℃.

After completion, additive A is prepared to proceed with pH adjustment and neutralization process. At this time, since the viscosity of the contents rises, stir vigorously (6,000 rpm) evenly for 5 minutes or more.

When the contents cool down below 50℃, add additive B and mix slowly for 2 minutes (3,000~4,000 rpm). After stirring, the contents are collected through a degassing/filtration process.

After completion, the pH adjustment and neutralization process proceeds. At this time, since the viscosity of the contents rises, stir vigorously (6,000 rpm) evenly for more than 5 minutes.

When the contents cool down below 50℃, add a preservative and slowly mix for 2 minutes (3,000~4,000rpm). When the temperature is below 40℃, add tetra peptide and flavoring, stir slowly for 2 minutes (3,000~4,000rpm), and degas /Collect the contents through the filtration process.

The prepared cream was stored at low temperature (4℃), high temperature (50℃), room temperature (25℃), and sunlight conditions to check the stability of the cream. Discoloration, odor change, phase change, and pH change of the cream for 4 weeks under all conditions. The same phenomenon was not found, and it was confirmed that the stability of the cream was maintained through this. In particular, the stability of the improved peptide was maintained, and as a result of HPLC analysis, it was confirmed that the content was maintained at 90% or more for 4 weeks under all conditions.

division Raw material name / ingredient name Ingredients / INCI Name parts by weight Award A DI WATER Water 39.03 GLYCERIN Glycerin 10.00 Keltrol-F Xanthan Gum 0.10 Adenosine Adenosine 0.04 Award B DI Water Water 30.00 beta-glucan beta-glucan 0.20 Carbopol 940 Carbomer 0.02 Paid Kalcol 6870 Cetearyl Alcohol 2.50 GMS-105 Glyceryl Stearates 1.50 Arlacel 165V Glyceryl Stearate, Peg-100 Stearate 1.20 Tween 80 Polysorbate 80 1.00 Arlacel 83 Sorbitan Sesquioleate 0.70 Phytosqualane Squalane 5.00 DUB MCTs Caprylic/Capric Triglycerides 3.00 Additive A DI Water Water 3.00 L-Arginine (pH 4.50 ~ 5.50) Arginine 0.50 Additive B 1,2-Hexanediol 1,2-Hexanediol 2.00 Additive C glucosyl rutin glucosyl rutin 0.01 Additive D (Fragrance) Perfume Fragrance 0.10 functional ingredient Palmitoyl-GQVS Palmitoyl-GQVS 0.1

Above, preferred embodiments of the present invention have been described in detail. The description of the present invention is for illustrative purposes, and those skilled in the art will understand that other specific forms can be easily modified without changing the technical spirit or essential features of the present invention.

Therefore, the scope of the present invention is indicated by the following claims rather than the above detailed description, and all changes or modifications derived from the meaning, scope and equivalent concept of the claims are included in the scope of the present invention. should be interpreted

Claims (7)

An improved peptide in which a peptide consisting of an amino acid sequence of Glycine (Gly)-Glutamine-Valine (Val)-Serine (Ser) and a saturated or unsaturated fatty acid having 10 to 20 carbon atoms are amide bonded. According to claim 1
The improved peptide, characterized in that the fatty acid is palmitic acid. An improved peptide comprising a peptide consisting of an amino acid sequence of Glycine (Gly)-Glutamine-Valine (Val)-Serine (Ser) and an acetyl group bonded to an amide bond. According to claim 1 or 3,
The improved peptide is an improved peptide, characterized in that synthesized using 2- (4-nitrophenyl) sulfonylethoxycarbonyl-amino acid (Nsc-amino acid) as an intermediate. According to claim 1 or 3,
The improved peptide is an improved peptide, characterized in that the thermal stability is improved compared to the peptide. According to claim 1 or 3,
The improved peptide is an improved peptide, characterized in that the protease resistance is improved compared to the peptide. A cosmetic composition comprising the improved peptide of claim 1 or 2.

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