Novel peptide and cosmetic composition comprising same

Abstract

The present invention relates to a novel peptide and derivative thereof that exhibit wrinkle reduction activity and a cosmetic composition comprising same and having an excellent wrinkle reduction effect. The novel peptide and derivative thereof of the present invention exhibit an excellent wrinkle reduction effect, are easily absorbed due to the short amino acid sequence length thereof, and have few side effects, and therefore can be widely used in a cosmetic composition for improving skin condition.

Description

Novel peptide and cosmetic composition containing the same

[0001] The present invention relates to a novel peptide and its derivative exhibiting wrinkle improvement activity, and a cosmetic composition containing the same having an excellent wrinkle improvement effect.

[0002] The skin, as the body's primary defense barrier, protects internal organs from external environmental stimuli such as temperature and humidity changes, ultraviolet rays, and pollutants, and plays a crucial role in maintaining homeostasis, including body temperature regulation. However, excessive physical and chemical stimuli and stress applied from outside the skin cells, nutritional deficiencies, ultraviolet rays, environmental pollutants, and rapid temperature changes can induce various biochemical changes within the cells, accelerating skin aging, such as freckles, blemishes, loss of elasticity, keratinization, and wrinkles.

[0003] Aging causes wrinkles on the skin, and major factors contributing to wrinkle formation include UV exposure and decreased collagen synthesis. Skin aging can be broadly divided into intrinsic aging, driven by genetic factors, and extrinsic aging, driven by external environmental factors like sunlight. Among these, extrinsic aging is known to be preventable, treatable, or even delayed by eliminating free radicals, promoting fibroblast proliferation, and stimulating collagen synthesis.

[0004] Against this technological backdrop, multifaceted research is being conducted on peptides and other substances that can improve skin condition.

[0005] Accordingly, the inventors of the present invention have conducted research and efforts to develop a peptide material that can be used in a cosmetic composition, and as a result, have discovered that a novel synthetic peptide and its derivative exhibit excellent wrinkle improvement effects, thereby completing the present invention.

[0006] Accordingly, the purpose of the present invention is to provide a novel peptide and its derivatives and a cosmetic composition for improving wrinkles comprising the same.

[0007] The present invention,

[0008] A peptide represented by the amino acid sequence of the following chemical formula 1, a stereoisomer thereof, a solvate thereof, a salt thereof, or a co-former thereof is characterized.

[0009] [Chemical Formula 1]

[0010] R1-W n -X m -AAQP-Y p -Z q -R2

[0011] W, X, Y, Z are independently amino acids,

[0012] n, m, p, q are independently 0 or 1,

[0013] R1 is H, C1-C 24 Alkyl, C1-C 24 alkenyl, C1-C 24 alkynyl or R3-CO-, wherein R3 is H or C1-C 24 alkyl, C1-C 24 alkenyl or C1-C 24 It is an alkynyl,

[0014] R2 is -NR4R5 or -OR6, where R4, R5 and R6 are independently H or C1-C 24 Alkyl, C1-C 24 Alkenyl or C1-C 24 It's alkynyl.

[0015]

[0016] In the above chemical formula 1,

[0017] W is glutamic acid (E) or arginine (R),

[0018] X is glutamic acid (E), glycine (G), arginine (R), glutamine (Q), or alanine (A),

[0019] Y is serine (S), proline (P), arginine (R), or glycine (G),

[0020] Z can be glutamine (Q), threonine (T), or proline (P).

[0021]

[0022] In addition, the present invention is characterized by a peptide represented by the following chemical formulas 2 to 10, a stereoisomer thereof, a solvate thereof, a salt thereof, or a co-former thereof.

[0023] [Chemical Formula 2]

[0024] AAQP

[0025]

[0026] [Chemical Formula 3]

[0027] CH3CO-AAQP

[0028]

[0029] [Chemical Formula 4]

[0030] AAQP-NHC2H5

[0031]

[0032] [Chemical Formula 5]

[0033] EAAQP

[0034]

[0035] [Chemical Formula 6]

[0036] AAQPS

[0037]

[0038] [Chemical Formula 7]

[0039] RGAAQP-NH2

[0040]

[0041] [Chemical Formula 8]

[0042] CH3CO-RAAQPPQ

[0043]

[0044] [Chemical Formula 9]

[0045]

[0046] QAAQPRT

[0047]

[0048] [Chemical Formula 10]

[0049] EAAAQPGP

[0050]

[0051] Meanwhile, the present invention is characterized by a cosmetic composition comprising the peptide, a stereoisomer thereof, a solvate thereof, a salt thereof, or a co-former thereof.

[0052] The above cosmetic composition is characterized by an excellent wrinkle improvement effect.

[0053] The novel peptide of the present invention and its derivatives have excellent wrinkle improvement effects, are easy to absorb due to their short amino acid sequence length, and have few side effects, so they can be widely used in cosmetic compositions for improving skin conditions.

[0054] Figure 1 is a graph showing the results of cytotoxicity evaluation of the example peptide.

[0055] Figure 2 is a graph showing the amount of procollagen type 1 peptide produced by the example peptide.

[0056] Hereinafter, the present invention will be described in detail. Prior to this, it should be noted that the terms and words used in this specification and claims should not be interpreted as limited to their conventional or dictionary meanings. Rather, they should be interpreted with meanings and concepts that conform to the technical spirit of the present invention, based on the principle that the inventor can appropriately define the concept of the term to best explain his or her invention. Accordingly, the configurations described in the embodiments described in this specification are merely the most preferred embodiments of the present invention and do not represent the entire technical spirit of the present invention. Therefore, it should be understood that various equivalents and modified examples may exist as of the time of this application.

[0057]

[0058] The novel peptide of the present invention is represented by the following chemical formula 1, a stereoisomer thereof, a solvate thereof, a salt thereof, or a co-former thereof.

[0059] [Chemical Formula 1]

[0060] R1-W n -X m -AAQP-Y p -Z q -R2

[0061]

[0062] W, X, Y, Z are independently amino acids,

[0063] n, m, p, q are independently 0 or 1,

[0064] R1 is H, C1-C 24 Alkyl, C1-C 24 alkenyl, C1-C 24 alkynyl or R3-CO-, wherein R3 is H or C1-C 24 Alkyl, C1-C 24 Alkenyl or C1-C 24 It is alkynyl,

[0065] R2 is -NR4R5 or -OR6, where R4, R5 and R6 are independently H or C1-C 24 Alkyl, C1-C 24 Alkenyl or C1-C 24 It's alkynyl.

[0066]

[0067] The amino acids mentioned in this specification are abbreviated as follows according to the IUPAC-IUB nomenclature.

[0068] A: Alanine (Ala)

[0069] Q: Glutamine (Gln)

[0070] P: Proline (Pro)

[0071] (Sequence number 1: AAQP)

[0072]

[0073] In the present invention, the term “peptide or fragment thereof” means a polymer composed of two or more amino acids linked by amide bonds (or peptide bonds).

[0074] In the above chemical formula 1, the hyphen (-) indicating a peptide bond removes the OH of the 1-carboxyl group of the amino acid (represented herein in its conventional non-ionized form) when it is located to the right of the symbol, and removes the H of the 2-amino group of the amino acid when it is located to the left of the symbol, and both of these modifications can be applied to the same symbol.

[0075] The above peptides may have a protecting group attached to the N- or C-terminus of the peptide to obtain chemical stability, enhanced pharmacological properties (half-life, absorbability, potency, efficacy, etc.), altered specificity (e.g., broad biological activity spectrum), and reduced antigenicity. In one specific example, the N-terminus of the peptide (wherein the hyphen is located on the left side of the symbol) may be bonded to any one protecting group selected from the group consisting of a formyl group, an acetyl group, a benzoyl group, a fluoreonylmethoxycarbonyl group, a palmitoyl group, a butoxycarbonyl group, an aryloxycarbonyl group, a methyl group, an ethyl group, a benzyl group, a myristyl group, and a stearyl group, and the C-terminus of the peptide (wherein the hyphen is located on the right side of the symbol) may be bonded to any one protecting group selected from the group consisting of an amino group (-NH2), an alkyl amino group, an azide (-NHNH2), a methoxy, an ethoxy, a propoxy, and a polyethylene glycol. It can be combined with any one protecting group selected from the group consisting of glycol; PEG).

[0076] C1-C above 24 Alkyl may be composed of, but is not limited to, methyl, ethyl, isopropyl, isobutyl, tert-butyl, heptyl, octyl, decyl, dodecyl, lauryl, hexadecyl, octadecyl, amyl, 2-ethylhexyl, 2-methylbutyl, 5-methylhexyl, and the like.

[0077] C1-C above 24Alkenyl may be composed of, but is not limited to, a vinyl group, an allyl group, a 1-propenyl group, an isopropenyl group, a 1-butenyl group, a 2-butenyl group, a 3-butenyl group, a 1-pentenyl group, a 2-pentenyl group, a 3-pentenyl group, etc.

[0078] C1-C above 24 Alkynyl may be composed of, but is not limited to, ethynyl, 2-propynyl, etc.

[0079] The above W, X, Y, and Z are independently amino acids, wherein W is glutamic acid (E) or arginine (R), X is glutamic acid (E), glycine (G), arginine (R), glutamine (Q), or alanine (A), Y is serine (S), proline (P), arginine (R), or glycine (G), and Z may be glutamine (Q), threonine (T), or proline (P).

[0080] However, the case where the above n, m, q are 0, p is 1, Y is arginine (R), R1 is H, and R2 is OH is excluded from the peptide of the present invention.

[0081] Preferably, the peptide of the present invention may be a compound represented by the following chemical formulas 2 to 10.

[0082]

[0083] [Chemical Formula 2]

[0084] AAQP

[0085]

[0086] [Chemical Formula 3]

[0087] CH3CO-AAQP

[0088]

[0089] [Chemical Formula 4]

[0090] AAQP-NHC2H5

[0091]

[0092] [Chemical Formula 5]

[0093] EAAQP

[0094] (Sequence number 2: EAAQP)

[0095]

[0096] [Chemical Formula 6]

[0097] AAQPS

[0098] (Sequence number 3: AAQPS)

[0099]

[0100] [Chemical Formula 7]

[0101] RGAAQP-NH2

[0102] (Sequence number 4: RGAAQP)

[0103]

[0104] [Chemical Formula 8]

[0105] CH3CO-RAAQPPQ

[0106] (Sequence number 5: RAAQPPQ)

[0107]

[0108] [Chemical Formula 9]

[0109] QAAQPRT

[0110] (Sequence number 6: QAAQPRT)

[0111]

[0112] [Chemical Formula 10]

[0113] EAAAQPGP

[0114] (SEQ ID NO: 7: EAAAQPGP)

[0115]

[0116] The peptides of the present invention may exist as stereoisomers or mixtures of stereoisomers, for example, the amino acids constituting them may independently be D-form, L-form, or D,L-form (racemate). Accordingly, depending on the number of asymmetric carbon atoms and which isomer or mixture of isomers is present, not only isomer mixtures but also racemic mixtures, diastereomeric mixtures, or pure diastereoisomers or enantiomers can be obtained. The preferred structure of the peptides of the present invention is a pure isomer, i.e., an enantiomer or diastereoisomer.

[0117] The above solvate means the peptide of the present invention or a salt thereof containing a stoichiometric or non-stoichiometric solvent bound by non-covalent intermolecular forces, and the solvent may be water, in which case the solvate is referred to as a hydrate.

[0118] Furthermore, the salt means a salt recognized for its use in animals, more specifically in humans, and may include base addition salts (e.g., inorganic salts such as lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc or aluminum salts, organic salts such as ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, arginine, lysine, histidine or piperazine salts), or acid addition salts (e.g., organic salts such as acetate, citrate, lactate, malonate, maleate, tartrate, fumarate, benzoate, aspartate, glutamate, succinate, oleate, trifluoroacetate, oxalate, pamoate or gluconate salts, inorganic salts such as chloride, sulfate, borate or carbonate salts).

[0119] In addition, the co-former is not particularly limited as long as it is pharmaceutically or cosmetically acceptable, and may be at least one selected from meglumine, histidine, arginine, nicotinamide, benzoate, formic acid, sorbic acid, citric acid, malic acid, caffeine, theophylline, and urea.

[0120] The peptide of the present invention can be synthesized by a method well known in the art, for example, an automatic peptide synthesizer, and can also be produced by genetic engineering techniques, but is not limited thereto.

[0121] Additionally, the present invention provides a polynucleotide encoding the peptide. By inserting the polynucleotide encoding the peptide into a vector and expressing it, the peptide can be obtained in large quantities.

[0122]

[0123] Meanwhile, the present invention provides a cosmetic composition comprising the peptide, a stereoisomer thereof, a solvate thereof, a salt thereof, or a co-former thereof.

[0124] The above cosmetic composition can be used particularly for wrinkle improvement purposes.

[0125] The cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions, and may include conventional auxiliary agents such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances, and carriers.

[0126] The cosmetic composition of the present invention can be prepared in any formulation commonly manufactured in the art, and can be formulated as, for example, a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation, and a spray, but is not limited thereto.

[0127] More specifically, it can be manufactured in the form of a flexible toner, a nourishing toner, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder.

[0128] When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component.

[0129] When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether may be additionally included.

[0130] When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

[0131] When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, etc. can be used as a carrier component.

[0132] When the formulation of the present invention is a surfactant-containing cleansing agent, a fatty alcohol sulfate, a fatty alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyl taurate, a sarcosinate, a fatty acid amide ether sulfate, an alkyl amidobetaine, a fatty alcohol, a fatty acid glyceride, a fatty acid diethanolamide, a vegetable oil, a lanolin derivative, or an ethoxylated glycerol fatty acid ester may be used as a carrier component.

[0133] When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation, or a surfactant-free cleansing formulation, it can be applied to the skin and then wiped off, removed, or washed off with water. As specific examples, the soap is liquid soap, powder soap, solid soap, and oil soap, the surfactant-containing cleansing formulation is cleansing foam, cleansing water, cleansing towel, and cleansing pack, and the surfactant-free cleansing formulation is cleansing cream, cleansing lotion, cleansing water, and cleansing gel, but is not limited thereto.

[0134]

[0135] Hereinafter, the present invention will be described in detail using examples and experimental examples. However, the examples according to the present invention may be modified in various ways, and the scope of the present invention should not be construed as being limited to the examples described below. The examples of the present invention are provided to more fully explain the present invention to those of average skill in the art.

[0136]

[0137] Example: Production of peptides

[0138] Peptides having the amino acid sequences of Examples 1 to 9 below were synthesized using an automatic peptide synthesizer (PetiSyzer®, Samjeon Pure Chemical Industries, Korea). The purity of the synthesized peptides was confirmed using high-performance liquid chromatography (HPLC, SHIMADZU Prominence, Japan), and the molecular weight was confirmed through mass analysis (SHIMADZU LC-MS-8045). The results are shown in Table 1. The column used for the analysis was ACQUITY UPLC® BEH C18 (50 mm, 1.7 μm, Waters, USA). The mobile phase was distilled water containing 0.1% trifluoroacetic acid (TFA) and acetonitrile containing 0.1% trifluoroacetic acid (TFA). The flow rate was 1.0 ml / min, and an ultraviolet spectrophotometer (214 nm) was used as the detector. The elution conditions were measured in gradient mode by changing the ratio of acetonitrile containing 0.1% TFA (trifluoroacetic acid) for 15 minutes.

[0139] No. Peptide sequence purity (%) Molecular weight (g / mol) Example 1 AAQP 96.93 86.2 Example 2 Ac-AAQP 96.94 27.21 Example 3 AAQP-NHEt 99.02 412.24 Example 4 EAAQP 97.42 514.24 Example 5 AAQPS 98.03 472.23 Example 6 RGAAQP-NH 2 96 597.33 Example 7 Ac-RAAQPPQ 96.77 808.42 Example 8 QAAQPRT 98.84 770.4 Example 9 EAAAQPGP 97.98 668.31

[0140] Test Example 1: Cell Culture

[0141] Dermal fibroblast culture

[0142] Normal Human Dermal Fibroblasts (NHDF) cells, which are dermal fibroblasts, were obtained from Lonza (Lonza Walkersville, Inc.). They were cultured in Fibroblast Basal Medium (FBM) containing 0.1% hFGF-B, insulin, GA-1000, and 2% fetal bovine serum at 37°C in a 5% CO2 incubator, and passaged at 2-3 day intervals.

[0143]

[0144] Test Example 2: Cytotoxicity Evaluation

[0145] EZ-cytox assay

[0146] The EZ-cytox assay is a representative method for measuring cell viability that utilizes the principle that WST (water solution tetrazolium salt) reacts with the dehydrogenase of living cells to produce orange, water-soluble formazan.

[0147]

[0148] Cytotoxicity evaluation on skin fibroblasts

[0149] To determine cytotoxicity, NHDF cells were cultured in 1.0×10 4 Cells were seeded into a 96-well plate at 10 cells / well and cultured for 24 hours at 37°C and 5% CO2. The cultured cells were exchanged for serum-free FBM and treated with the samples to be evaluated. After that, EZ-cytox was added to each well and reacted for 30 minutes at 37°C and 5% CO2, and the absorbance was measured at 450 nm using a microplate reader. The average absorbance value for each sample group was obtained, and cell viability was evaluated by comparing this value with the absorbance value of the control group, and the results are shown in Table 2 and Fig. 1. No cytotoxicity was observed at the concentrations evaluated.

[0150] Concentration (μM) cell viability (%) SD (%) Control 100.0000.000 Example 1 100.96.48 5.013 Example 2 100.100.87 73.200 Example 3 100.89.02 05.148 Example 4 100.98.103 2.329 Example 5 100.98.75 52.976 Example 6 100.97.13 73.251 Example 7 100.90.75 70.468 Example 8 100.96.5 202.868 Example 9 100.97.09 20.509 Ascorbic acid (μg / mL) 1090.67 41.293

[0151] Test Example 3: Confirmation of the production of procollagen type I peptide (PIP), a wrinkle-improving factor.

[0152] NHDF cells were seeded at 2.0×10 in a 24-well plate. 4 The cells were seeded at 100 cells / well and cultured for 24 hours at 37°C and 5% CO2 conditions. The cultured cell medium was replaced with serum-free FBM medium, and the peptide of the above example was treated at a concentration of 100 μM and cultured for 24 hours. Afterwards, the supernatant of each experimental group was taken and the amount of free procollagen in the medium was measured. The control group was treated with ascorbic acid at a concentration of 0.001 v / v%. The measurement of the amount of procollagen was performed using a procollagen type I peptide (PIP) EIA kit (Takara Biomedical Co.) according to the method provided by the manufacturer. The quantitative results of the production of procollagen type I peptide according to the treatment of the above example are shown in Table 3 and Fig. 2 below.

[0153] Concentration (μM) PIP production (%) SD (%) Control 100.000 11.64 2 Example 1 100 139.53 9 10.41 2 Example 2 100 130.84 32.75 4 Example 3 100 153.79 84.80 3 Example 4 100 138.05 5 1.93 8 Example 5 100 136.34 6 2.90 6 Example 6 100 137.58 13.01 4 Example 7 100 139.47 8 0.57 5 Example 8 100 144.59 10.25 3 Example 9 100 144.86 40.59 1 Ascorbic acid (μg / mL) 101 72.88 42.52 3

[0154] As shown in Table 3 and Figure 2 above, when the peptide of the example was treated, the amount of pro-collagen production increased, confirming that the peptide of the present invention exhibits excellent wrinkle improvement and anti-aging effects.

Claims

1. A peptide represented by the amino acid sequence of the following chemical formula 1, a stereoisomer thereof, a solvate thereof, a salt thereof, or a co-former thereof: [Chemical Formula 1] R1-W n -X m -AAQP-Y p -Z q -R2 W, X, Y, Z are independently amino acids, n, m, p, q are independently 0 or 1, R1 is H, C1-C 24 Alkyl, C1-C 24 alkenyl, C1-C 24 alkynyl or R3-CO-, wherein R3 is H or C1-C 24 Alkyl, C1-C 24 Alkenyl or C1-C 24 It is alkynyl, R2 is -NR4R5 or -OR6, where R4, R5 and R6 are independently H or C1-C 24 Alkyl, C1-C 24 Alkenyl or C1-C 24 It's alkynyl.

2. In paragraph 1, W is glutamic acid (E) or arginine (R), X is glutamic acid (E), glycine (G), arginine (R), glutamine (Q), or alanine (A), Y is serine (S), proline (P), arginine (R), or glycine (G), A peptide, a stereoisomer thereof, a solvate thereof, a salt thereof or a copolymer thereof, wherein Z is glutamine (Q), threonine (T) or proline (P).

3. In paragraph 1, A peptide represented by the following chemical formulas 2 to 10, a stereoisomer thereof, a solvate thereof, a salt thereof, or a co-former thereof: [Chemical Formula 2] AAQP [Chemical Formula 3] CH3CO-AAQP [Chemical Formula 4] AAQP-NHC2H5 [Chemical Formula 5] EAAQP [Chemical Formula 6] AAQPS [Chemical Formula 7] RGAAQP-NH2 [Chemical Formula 8] CH3CO-RAAQPPQ [Chemical Formula 9] QAAQPRT [Chemical Formula 10] EAAAQPGP 4. A cosmetic composition comprising the peptide of paragraph 1, a stereoisomer thereof, a solvate thereof, a salt thereof, or a co-former thereof.

5. In paragraph 4, The above composition is a cosmetic composition characterized by having an excellent wrinkle improvement effect.

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