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WO2012128464A1 - Dérivés dipeptidiques d'undécénoyle et composition cosmétique de blanchiment de la peau en contenant - Google Patents

Dérivés dipeptidiques d'undécénoyle et composition cosmétique de blanchiment de la peau en contenant Download PDF

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WO2012128464A1
WO2012128464A1 PCT/KR2012/000697 KR2012000697W WO2012128464A1 WO 2012128464 A1 WO2012128464 A1 WO 2012128464A1 KR 2012000697 W KR2012000697 W KR 2012000697W WO 2012128464 A1 WO2012128464 A1 WO 2012128464A1
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undecenoyl
ome
examples
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melanin
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Korean (ko)
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이윤섭
최하영
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MIWON COMMERCIAL CO Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06026—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
    • C07K5/06043—Leu-amino acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06078—Dipeptides with the first amino acid being neutral and aromatic or cycloaliphatic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06139—Dipeptides with the first amino acid being heterocyclic
    • C07K5/06156—Dipeptides with the first amino acid being heterocyclic and Trp-amino acid; Derivatives thereof

Definitions

  • the present invention relates to an undecenoyl dipeptide derivative and a cosmetic composition for skin whitening comprising the same, and more particularly, acts as an antagonist of MC1R, an alpha melanocyte stimulating hormone ( ⁇ -MSH) receptor, leading to skin blackening. It binds to its receptor, MC1R, through competitive binding with its alpha melanocyte stimulating hormone, which blocks melanin synthesis pathways and effectively blocks inflammation-related signaling pathways, resulting in undecenoyl depeptides with excellent skin whitening and anti-inflammatory effects. It relates to a derivative and a cosmetic composition for skin whitening comprising the same.
  • Melanin is produced in the melanocytes (Melanosome), which are organelles that exist in cells called melanocytes (Melanocytes) in the basement layer of the skin. The amount is determined. The production of melanin is determined by genetic factors such as congenital factors, female hormones, drugs, pregnancy, ultraviolet rays, stress, etc., which are most influenced by UV rays. Melanin absorbs the light energy of ultraviolet rays and protects cells from damage caused by ultraviolet rays. When abnormally low melanin is produced, skin lesions such as vitiligo are caused, and excessive production causes damage to the skin as well as the formation of blemishes and freckles, and even skin cancer.
  • melanocytes melanocytes
  • UV rays ultraviolet rays
  • the skin of modern man is damaged by exposure to environmental pollution, external stimuli such as ultraviolet rays and stress.
  • external stimuli such as ultraviolet rays and stress.
  • the damaged skin has a slow recovery rate, wrinkles, decreased elasticity and skin pigmentation.
  • the expectation for the whitening effect is very high, and various cosmetics have been developed to reduce the color of melanin already produced or to suppress the production of new melanin.
  • melanocyte stimulating hormone ⁇ -MSH
  • endothelin-1 endothelin-1
  • ⁇ -MSH When the ⁇ -MSH binds to the melanocortin-1 receptor (MC1R), which is an ⁇ -MSH receptor present in the melanocyte membranes, by stimulating melanin forming cells such as ultraviolet rays, hormones, and drugs, the stimulated MC1R Activates an enzyme called adenylate cyclase, which increases cyclic adenosine monophosphate (cAMP) in the cell and causes a series of processes of melanin biosynthesis to occur continuously by increased cAMP. do. cAMP sequentially activates protein kinase A (PKA) and CREB transcription factors to promote expression of MITF (Microphthalmia associated transcription factor), a precursor of ⁇ -MSH.
  • M1R melanocortin-1 receptor
  • MITF causes transcriptional activity of melanin synthesis-related genes such as tyrosinase, tyrosinase related protein 1 (TRP1) and dopachrome tautomerase (DCT), tyrosine in melanocytes (tyrosine) in the melanocytes (tyrosinase; Important enzyme) is converted into dopa (DOPA; 3,4-dihydroxyphenylalanine) and dopaquinone.
  • TRP1 tyrosinase related protein 1
  • DCT dopachrome tautomerase
  • Endothelin-1 is also increased by stimulation of ultraviolet rays, hormones, drugs, and the like, and binds to endothelin B type receptor (ETbR) present in melanocyte membranes.
  • Phospholipase C (PLC) stimulated by ETbR is used to convert phosphatidylinositol 4,5-bisphosphate (PIP 2 ) present in the cell membrane into diacyl glycerol (DAG) and inositol. Decompose with 1,4,5-trisphosphate (inositol 1,4,5-trisphosphate: IP 3 ).
  • IP 3 binds to the IP 3 receptors of the endoplasmic reticulum, opening up the calcium channel of the endoplasmic reticulum, and increasing the calcium concentration in the cytoplasm is involved in melanin synthesis through several chain reactions in the cell.
  • PKC Protein kinase C
  • Endoderin activates PKA, as well as PKC, through cAMP stimulation. It is known that melanogenesis occurs through the cross-talk of these two kinase enzymes.
  • Endothelin-1 / ETbR signaling is also synergistic with SCF / c-KIT, another pathway involved in melanin synthesis, increasing tyrosinase expression, leading to skin blackening.
  • pheomelanin phemelanin (red melanin) is formed), or through enzymatic reactions such as TRP1, DCT, eumelanin (eumelanin (melanin, black or brown melanin) is formed.
  • the produced melanin is transferred from the melanocytes to the keratinocytes (keratinocytes), which form the epidermis, through the melanosomes, and is deposited on the skin when overproduced according to the amount of melanin produced in this process. It will cause freckles and skin blackening.
  • a substance having a tyrosinase inhibitory activity such as arbutin, ascorbic acid, kojic acid, glutathione, and the like is added to the cosmetic to give a skin whitening effect, It has improved skin hyperpigmentation such as blemishes and freckles.
  • kojic acid effectively inhibits enzymatic activity by adsorbing copper ions present in the active site of tyrosinase, but it is unstable when blended into cosmetics and causes skin troubles.
  • Ascorbic acid and its derivatives have a poor skin penetration. It is difficult, and its efficacy is poor when used as a cosmetic raw material because of the oxidation instability.
  • Hydroquinone causes allergies, high cytotoxicity and irritation and is limited in each country.
  • Arbutin is a derivative in which glucopyranoside is bound to hydroquinone, which has been widely used because it has no side effects of hydroquinone, and has been spotlighted, but it is decomposed by skin enzymes. That is, the existing whitening raw materials are severe skin irritation or toxic, the product stability is not good, the skin penetration is not good, the whitening effect is insignificant, there is a limit in use.
  • ETbR stimulation Another cause of melanin biosynthesis is ETbR stimulation.
  • endodelin-1 When endodelin-1 is secreted by ultraviolet rays, it promotes the secretion of various inflammation-related cytokines (Cytokine) as well as skin blackening caused by ETbR stimulation.
  • the endothelin-1 / ETbR stimulatory signal stimulates the secretion of cytokines such as IL-1a, IL-1b, COX-1, COX-2, TNF- ⁇ , IFN- ⁇ , and PEG2, causing UV-inflammation.
  • N-cadherin (M-Cherin) MMP-1, MMP-2, MMP-9, Integrin (Integrin) to increase the expression of cancer cells to help metastasis.
  • the development of ETbR antagonists will have the function of inhibiting the skin inflammatory response caused by ultraviolet light and protect skin cells from skin cancer.
  • the whitening effect is excellent, as well as stabilizing the damaged skin due to ultraviolet rays, excellent skin permeability or stability, bio-friendly development of a non-toxic raw material has emerged as an important key.
  • the present inventors focused on MC1R, which plays the most important role in skin pigmentation process, and synthesized an antagonist of MC1R, and thus the undecylenoyl dipeptide derivatives according to the present invention was applied to skin blackening caused by MC1R.
  • the inhibitory effect was demonstrated to be very good, the signal transduction process was elucidated, and ETbR blackening stimulation was also demonstrated to be inhibited through MC1R-dependent cross-reaction.
  • ETbR blackening stimulation was also demonstrated to be inhibited through MC1R-dependent cross-reaction.
  • Undecenoyl dipeptide derivatives used in the present invention are arbutin, kojic acid, which is widely used as a whitening test material, and DermaPep TM UL (patent application number PCT / KR2010 / 006387, INCI name: Methyl Undecenoyl)
  • the whitening effect was better than that of Leucinate.
  • the present inventors have developed a functional whitening material with an anti-inflammatory effect and proved its efficacy.
  • an object of the present invention is to provide an undecenoyl dipeptide derivative which is harmless to the human body, has an excellent whitening effect and is accompanied by an anti-inflammatory effect, and has excellent skin permeability and stability, and a cosmetic composition for skin whitening containing the same.
  • the present invention provides an undecenoyl dipeptide derivative.
  • the undecenoyl dipeptide derivative according to the present invention is represented by the following Chemical Formula 1.
  • -C ( 0)-means carbonyl
  • a 1 , A 2 are amino acid residues selected from Leu, Phe, Trp and Gly, respectively, where Leu, Phe and Trp are L- Or a D-form or a mixture thereof
  • R is H, C 1 -C 18 alkyl, phenyl, or benzyl
  • the undecenoyl group is bonded to an amine group of A 1
  • OR is a carbon of A 2 To bind to a carbonyl group.
  • a 1 and A 2 are composed of L- or D-type or a mixed form of leucine, and R is preferably a methyl group, and more preferably, the undecenoyl dipeptide derivative is an undeceno.
  • L means l-type leucine
  • L means d-type leucine.
  • the present invention also provides a cosmetic composition for skin whitening comprising the above-described undecenoyl dipeptide derivative as an active ingredient.
  • the undecenoyl dipeptide derivative is preferably 0.0001 to 10% by weight based on the total weight of the cosmetic composition for skin whitening.
  • the cosmetic composition for skin whitening may be made of any one formulation selected from the group consisting of lotions, creams, foundations, essences, gels, packs, foam cleansing, and soap.
  • the present invention also provides a cosmetic composition for preventing inflammatory diseases associated with ultraviolet rays, comprising the above-described undecenoyl dipeptide derivative as an active ingredient.
  • the undecenoyl dipeptide derivative acts as an antagonist of MC1R, effectively inhibits the expression of genes in the lower stage, inhibits the activity of tyrosinase, and inhibits the major enzyme of melanogenesis. It has the effect of inhibiting melanin production by inhibiting expression.
  • the undecenoyl dipeptide derivative is an MC1R antagonist and shows inhibitory effect on ETbR stimulation.
  • the undecenoyl dipeptide derivatives use biocompatible amino acids, resulting in low cytotoxicity and no skin irritation.
  • the undecenoyl dipeptide derivative has a low molecular weight and high skin penetration due to the organic affinity of fatty acids.
  • the present invention provides an undecenoyl dipeptide derivative.
  • the undecenoyl dipeptide derivative according to the present invention is represented by the following Chemical Formula 1.
  • -C ( O)-means carbonyl
  • a 1 , A 2 are amino acid residues selected from Leu, Phe, Trp and Gly, respectively, where Leu, Phe and Trp are L- Or a D-form or a mixture thereof
  • R is H, C 1 -C 18 alkyl, phenyl, or benzyl
  • the undecenoyl group is bonded to an amine group of A 1
  • OR is a carbon of A 2 To bind to a carbonyl group.
  • the undecenoyl dipeptide derivative of the present invention can be used as an active ingredient of a cosmetic composition that inhibits the production of melanin in cells and gives a whitening effect on the skin. Since the undecenoyl dipeptide derivative of the present invention acts as an antagonist of MC1R, a lotion, cream, essence, gel, ointment, pack, stick, powder, and the like, such as skin lotion and milk lotion, are known by a known cosmetic preparation method. Whitening cosmetics can be prepared in the form, and pharmaceutically acceptable carriers, excipients, and diluents can be added to the quasi-drugs and external medicines without any side effects.
  • Undecenoyl dipeptide derivatives according to the present invention can be synthesized through solid phase synthesis or solution synthesis.
  • solid phase synthesis the N-terminus is protected and the reactive side chains are protected.
  • the protecting group for protecting the N-terminus 9-fluorenylmethyloxycarbonyl group (Fmoc) is generally used.
  • solution synthesis conventionally well-known synthesis methods can be used.
  • the undecenoyl dipeptide methyl ester can be synthesized using a synthetic route as in Scheme 1.
  • Undecenoyl chloride (Undecenoyl chloride) and the amino acid (A 1 -OH) in the KOH aqueous solution as shown in Scheme 1 above (1) can be obtained undecenoyl amino acid (Undecenoyl amino acid).
  • the carbonyl group of the undecenoyl chloride reacts with the amine group of the amino acid to bind, and OH of A 1 -OH is OH of the carboxyl group of the amino acid.
  • Coupling Reagents that can be used at this time can be used as a general coupling agent widely used in peptide synthesis, for example, dicyclohexyl carbodiimide (DCC), HBTU (N-[(1H- benzotriazol-1-yl) (dimethylamino) methylene] -N-methylmethanaminium hexafluorophosphate N-oxide) or HATU (N-[(dimethylamino) -1-1H-1,2,3-triazolo [4,5-b] pyridin- 1-yl-methylene] -N-methylmethanaminium hexafluorophosphate N-oxide may be used, and HOAt (1-Hydroxy-7-Azabenzotriazole) or H
  • DCC dicyclohexyl carbodiimide
  • HBTU N-[(1H- benzotriazol-1-yl) (dimethylamino) methylene
  • the content of the undecenoyl dipeptide derivative contained in the preparation of the composition for skin whitening according to the present invention is preferably 0.0001 to 10% based on the total weight of the composition in consideration of the skin whitening effect and economical efficiency, and more preferably 0.002 1.0 wt%.
  • Examples 1 to 23 are applied in the present inventors DermaPep TM UL (Patent Application No. PCT / KR2010 / 006387), which is one of Leucine, Tryptophan, Phenylalanine, or Glycine. Synthesized in combination. Where L is Leucine, F is Phenylalanine, G is Glycine, and W is tryptophan, and these four amino acids are in vitro in consideration of the structure showing whitening efficacy through prior experiments of the present inventors. It was proved and selected through experimental system. OH at the peptide C-terminus represents a free acid group, OMe represents a methyl ester group, (d) represents a D-form amino acid, and an amino acid without such a designation is l- Type amino acids.
  • Examples 1 to 23 The whitening efficacy of the undecenoyl dipeptide compounds of Examples 1 to 23 was tested. Skin blackening is due to the production of melanin in the skin's melanocytes, which are released into the epidermis. Therefore, if the melanogenesis and the mechanism of melanocytes blocking the production of melanin, melanin is not produced and skin blackening does not appear. In addition, since Examples 1 to 23 will function as antagonists of MC1R, it was predicted that by binding to MC1R instead of ⁇ -MSH, it would inhibit the downstream signaling process of MC1R.
  • the undecenoyl dipeptide derivatives of Examples 1 to 23 were pretreated with melanin-producing cells, and treated with ⁇ -MSH, which artificially synthesizes melanin, to determine how much inhibition of skin blackening ability of ⁇ -MSH was detected. It was.
  • the whitening effect was measured by measuring the inhibitory effect on tyrosinase enzymes involved in the main stage of melanin synthesis. In the demonstration of efficacy of whitening raw materials, measuring the inhibition rate of melanin synthesis and tyrosinase enzyme activity in melanocytes is widely used as a basic experiment.
  • Mouse melanoma B16F10 cells were used as melanocytes. Cells were cultured at 37 ° C., 5% CO 2 ⁇ using 90% Dulbecco's modified eagle's medium (DMEM, GibcoBRL, USA) medium mixed with 10% fetal bovine serum (FBS; Hyclone, Lagan, UT). Incubated at 95% atmosphere (Incubator; Thermo-scientific, USA). Penicillin G and streptomycin (Phenicilin G sodium, 100 units / L, streptomycin sulfate, 100 mg / L; GibcoBRL, USA) were used as antibiotics.
  • DMEM Dulbecco's modified eagle's medium
  • FBS fetal bovine serum
  • Penicillin G and streptomycin Penicillin G and streptomycin (Phenicilin G sodium, 100 units / L, streptomycin sulfate, 100 mg / L; GibcoBRL, USA) were used as
  • Table 2 is a table showing the results of measuring their cytotoxicity by treating various concentrations of Examples 1 to 23.
  • B16F10 melanin producing cells were placed at 1 ⁇ 10 5 cells / well (Final volume 3 ml), incubated for 24 hours in a 37 ° C., 5% CO 2 incubator, and then the medium of each well was removed and a new DMEM (10% FBS). Whitening efficacy samples were prepared for each well by concentration and pretreated for 30 minutes.
  • the cells were lysed by vortex at 10 minute intervals for 30 minutes, centrifuged at 4 ° C and 13,000 rpm for 30 minutes to remove supernatant, and cell pellets containing melanin were recovered and 100 ml of Ether / Isopropanol 1: 1 solution. Wash once. The supernatant was removed by centrifugation at 13,000 rpm for 10 minutes, dried at 60 ° C. for about 3 hours, and 150 ml of 2N NaOH added with 10% DMSO was added thereto to dissolve intracellular melanin at 60 ° C. for 1 hour. . 100 ml of the supernatant obtained by dissolving each well of a 96-well plate was measured at 405 nm using an ELISA reader.
  • DMSO a solvent in which the sample was dissolved
  • the positive control did not process the sample, but treated only ⁇ -MSH to maximize melanin production.
  • Melanin synthesis inhibition rate (%) was calculated by the following equation.
  • Table 3 is a chart measuring the melanin synthesis inhibition rate (%) of the samples.
  • Examples 1 to 23 which are undecenoyl dipeptide derivatives, showed a concentration-dependent melanin synthesis inhibitory effect as the prescription concentration was increased from 5 ppm to 30 ppm. It showed much better melanin inhibitory effect than arbutin or kojic acid, which is known as a conventional whitening cosmetic. As a result, it was confirmed that Examples 1 to 23 are excellent candidate materials as whitening raw materials.
  • Example 2 and 3 [dipeptide sequence LL], Examples 4, 5 [dipeptide sequence LF], Examples 8, 9 [dipeptide sequence FL], Examples 12, 13 [dipeptide sequence GL], Examples 18, 19 [dipeptide sequence G (d) L ], Example 20, 21 [dipeptide sequence F (d) L]).
  • melanin synthesis inhibitory effect was the best in the repeat sequence of leucine (Leucine), the whitening effect was excellent in the sample containing the D-form amino acid when comparing the leucine repeat sequences (Example 3, 15, 16, 17).
  • B16F10 melanocytes or HEMn-LP melanocytes
  • HEMn-LP melanocytes were added to 1 ⁇ 10 5 cells / well (Final volume 3 ml), incubated in 37 ° C., 5% CO 2 incubator for 24 hours, The medium from each well was removed and replaced with fresh DMEM.
  • Whitening efficacy samples were prepared for each well by concentration and pretreated for 30 minutes.
  • the cells were lysed by vortex at intervals of 10 minutes for 30 minutes, then centrifuged at 4 ° C and 13,000 rpm for 30 minutes to transfer the supernatant to a new 1.5 ml tube.
  • Total protein was quantified using BCA protein assay kit (PIERCE, USA), and 40 ml of each sample was placed in each well of a 96-well plate, followed by phosphate buffer (pH, 6.8). 160 ml of L-3,4-dihydroxyphenylalanine (L-DOPA) dissolved immediately in After reacting for 20 to 60 minutes at 37 ° C, absorbance was measured at 475 nm using an ELISA reader.
  • L-DOPA L-3,4-dihydroxyphenylalanine
  • DMSO a solvent in which a sample was dissolved instead of a sample
  • the positive control did not process the sample, but only ⁇ -MSH was used to maximize the activity of tyrosinase enzyme.
  • the inhibition rate of tyrosinase enzyme activity (%) was calculated by the following equation.
  • % Tyrosinase activity inhibition [(A-C) / (A-B)] x 100
  • Table 4 is a chart measuring the percent inhibition of tyrosinase activity of the samples.
  • Examples 1 to 23 which are undecenoyl dipeptide derivatives, showed an effect of inhibiting concentration-dependent tyrosinase enzyme activity as the prescription concentration was increased from 5 ppm to 30 ppm. It showed much better inhibitory activity of tyrosinase enzyme activity than arbutin or kojic acid, which is known as a conventional whitening cosmetic. As a result, it was confirmed that Examples 1 to 23 were excellent whitening candidates that inhibited melanin production as well as inhibiting tyrosinase enzyme activity, which is an enzyme essential for melanin production.
  • Example 2 and 3 [Dipeptide Sequence LL ], Examples 4, 5 [dipeptide sequence LF], Examples 8, 9 [dipeptide sequence FL], Examples 12, 13 [dipeptide sequence GL], Examples 18, 19 [dipeptide sequence G (d ) L], Example 20, 21 [dipeptide sequence F (d) L]).
  • tyrosinase enzyme activity was the best inhibitory effect in the leucine repeat sequence, and the whitening effect was excellent in the sample containing D-form amino acids when leucine repeat sequences were compared.
  • Examples 1 to 23 inhibited melanin synthesis through inhibition of tyrosinase enzyme activity. If so, we examined which intracellular signal transduction pathways inhibited tyrosinase enzyme activity and inhibited melanin synthesis in their whitening efficacy.
  • the major signaling mechanism that stimulates melanogenesis is initiated by an increase in the amount of cAMP in the cell through adenylate cyclase. Therefore, it was measured how much the amount of intracellular cAMP is inhibited by Examples 1 to 23.
  • Examples 1 to 23 really act as MC1R-specific antagonists to show whitening efficacy.
  • Examples 16 and 17, which had the highest whitening effect increased tyrosinase activity and melanin production when PKA, cAMP, cGMP, PKC, and ETbR stimuli, which are factors causing skin blackening, were increased in addition to MC1R stimulation, respectively.
  • Experiments were conducted to evaluate which signaling pathways showed whitening efficacy by measuring how much inhibition was performed.
  • the present inventors anticipated that the whitening raw materials of the present invention would be specifically targeted to MC-R-stimulating ⁇ -MSH, and thus, the whitening raw materials of the present invention were ETbR-stimulated, which is known to cross-react in MC1R and cells. In addition, it was predicted to show some inhibitory effect through intracellular mechanism. In addition, since the whitening raw materials of the present invention will bind to the MC1R receptor and block its lower signaling mechanism, when stimulating intracellular proteins (PKA, cAMP, cGMP, PKC) activated by MC1R stimulation, MC1R present in the cell membrane It could be expected that the whitening material bound to the lysate would not be effective.
  • PKA intracellular proteins
  • melanin-related genes such as tyrosinase, TRP1, and TRP2.
  • % Transcription inhibition was measured using RT-PCR.
  • the amount of cAMP released into the cells was measured using a cAMP ELISA kit (Assay Design, USA).
  • the amount of cAMP generated during cell signal transduction of skin blackening was measured by ELISA (Enzyme-Linked Immunosorbent Assay) and the results are shown in Table 5.
  • the amount of cAMP in the cells was normalized to the amount of protein for each sample measured using the BCA protein assay.
  • Examples 1 to 23 inhibited the amount of cAMP in the cell, thereby affecting tyrosinase enzyme activity, which is a lower signaling pathway, and inhibited melanin synthesis. Since Examples 1 to 23 are made of MC1R antagonists, it is obvious that adenylate cyclase activity, which is a MC1R subsignal, is inhibited, thereby reducing intracellular cAMP release.
  • the undecenoyl dipeptide derivative used in the present invention was synthesized by modifying DermaPep TM UL. It has already been demonstrated that DermaPep TM UL is an MC1R specific antagonist (Patent Application No. PCT / KR2010 / 006387). Therefore, Examples 16 and 17, which were the most effective among Examples 1 to 23, were selected to determine whether they exhibit whitening efficacy by acting as MC1R-specific antagonists. Different stimulation to stimulate melanin synthesis by stimulating melanocytes, and demonstrated how effectively the blocking of Examples 16, 17 (%) and tyrosinase enzyme activity inhibition (%).
  • ⁇ -MSH M1R stimulation
  • Endothelin (16-21) EbR stimulation
  • Forskolin PKA stimulation
  • 8-Bromo-cAMP cAMP stimulation
  • 8-Bromo-cGMP cGMP stimulation
  • PMA PLC stimulation
  • Inhibition rate (%) of melanin synthesis is shown in Table 6, and tyrosinase enzyme activity inhibition rate (%) is shown in Table 6, and the effect of Examples 16 and 17 on the increased melanin synthesis and tyrosinase enzyme activity by treatment.
  • Reagents used for melanin stimulation were treated at the following concentrations.
  • Examples 16 and 17 were the most excellent inhibition of melanin production and tyrosinase enzyme activity when the melanin production stimulation by MC1R.
  • Forskolin, 8-Bromo-cAMP, 8-Bromo-cGMP, PKA by PMA, Adenylate cyclase, cGMP, PKC stimulation is a sub-step of the signaling mechanism in which MC1R antagonist acts, Examples 16, 17 Skin blackening irritation could not be prevented.
  • the ETbR stimulatory response by Endothelin (16-21) also showed some inhibitory effect. This assumes that ETbR and MC1R share several molecules in the cell and cross-talk. As a result, the MC1R antagonists of the present invention cross-talk with the ETbR signaling system, and proved to bind to MC1R and show a whitening effect.
  • B16F10 melanin producing cells were placed in a 60 mm dish at 5 ⁇ 10 5 cells / well (Final volume 5 ml), incubated for 24 hours in 37 ° C., 5% CO 2 incubator, and then the medium of each well was removed and replaced with fresh DMEM. Whitening efficacy samples were prepared for each well by concentration and pretreated for 30 minutes.
  • PCR primers were prepared beta-actin, tyrosinase, TRP1, TRP2, MITF, POMC of the mouse, and amplified 30 times under conditions of 94 ° C 1 minute, 60 ° C 1 minute, 72 ° C 1 minute.
  • PCR products were loaded on 1% agarose gel and electrophoresed for 20 minutes and identified using EC3 UV illuminator (UVP, USA). MRNA quantified by densitometer was quantified by dividing by beta-actin value. Primer sequences are shown in Table 8.
  • Examples 1 to 23 generally inhibited the mRNAs of Tyrosinase and TRP1 and TRP2.
  • Examples 1 to 23 which are MC1R antagonists, bind to MC1R to inhibit melanin synthesis and tyrosinase enzyme activity. Inhibiting the mRNA of tyrosinase, TRP1, TRP2 genes, it was proved to inhibit the whitening effect through inhibition at the transcription level. Moreover, the inhibitory effect of Examples 16 and 17 was the most excellent among Examples 1-23.
  • Examples 1 to 23 which are MC1R antagonists, act as MC1R-specific antagonists (Tables 6 and 7), reduce the amount of cAMP in cells (Table 5), tyrosinase, Inhibition of transcription of skin blackening-related genes such as TRP1 and TRP2 (Table 9) shows that whitening efficacy is shown by inhibiting tyrosinase enzyme activity and melanin biosynthesis (Tables 3 and 4).
  • Example 16 After treating with Example 16, 17 and ⁇ -MSH and incubating for 48 hours, replacing the medium with fresh medium, Examples 16, 17 were no longer treated, and only ⁇ -MSH was continued to be treated.
  • Table 11 Examples 16, 17% inhibition of tyrosinase activity at recovery, 48 h after treatment division Treatment concentration (ppm) When sample is not collected, continue processing with ⁇ -MSH 48 hours later, only sample recovery ⁇ -MSH Negative Control (Sample-Free) DMSO 0.01% - - DermaPep TM UL 5 4 31 15 32 0 30 70 48
  • Example 16 Undecenoyl L (d) L -OMe 5 11 One 15 76 24 30 82 39
  • Example 17 Undecenoyl (d) L (d) L-OMe 5 3 -3 15 71 35 30 85 42
  • Human keratinocyte HaCaT cells were used as keratinocytes.
  • Cells were prepared at 90 ° Dulbecco's modified eagle's medium (DMEM, GibcoBRL, USA) medium containing 10% fetal bovine serum (FBS; Cyclone, Lagan, UT) at 37 ° C, 5% CO 2 -95% (Incubator; Thermo-scientific, USA).
  • Penicillin G and streptomycin (Phenicilin G sodium, 100 units / L, streptomycin sulfate, 100 mg / L; GibcoBRL, USA) were used as antibiotics.
  • UV-B 100mJ / cm 2
  • serum-free DMEM medium serum-free DMEM medium
  • the culture medium was removed, washed with 1 ml of PBS, and RNA was extracted using an easy-spin TM (DNA free) Total RNA extraction kit (Intron, Korea).
  • PCR Primer was prepared in human gapdh, Interleukin-1a (IL-1a), Interleukin-1b (IL-1b), Interleukin 6 (IL-6), Interleukin 8 (IL-8), 94 ° C 1 minute, After 5 times amplification at 52 ° C. 1 min, 72 ° C.
  • Examples 1 to 21 are different in degree, but overall, IL-1a, IL-1b, IL-6, and IL-8, which are inflammation-related cytokines (Cytokine) increased by UV-B. Was effectively suppressed.
  • the amount of protein was measured by ELISA (Enzyme-Linked Immunosorbent Assay) method.
  • UV-B 100mJ / cm 2
  • serum-free DMEM medium serum-free DMEM medium
  • the culture medium was removed, washed with 1 ml of PBS, 1 ml of PBS was added, each well was scraped with a cell scraper to separate cells, and then transferred to each 1.5 ml tube. Subsequently, the supernatant was removed by centrifugation at 4 ° C.
  • Strip for ELISA was inserted into an ELISA plate, coated for 30 minutes (0.1% Glutaraldehyde in PBS), washed three times (0.1% Tween20 in PBS), 100ml each sample was added and reacted at 37 ° C for 3 hours. After the washing was repeated three times, the blocking buffer was added 100ml each reaction for 1 hour at 37 ° C.
  • the primary antibodies IL-1 ⁇ and IL-6 (antibody titer 1: 1000, Santacruz, USA) diluted in blocking buffer were added to each well 100ml each reaction at 37 ° C for 3 hours, washed three times After that, the secondary antibody (anti-rabbit, titer 1: 2000, Santacruz, USA) was added 100ml each reaction was performed at 37 ° C for 1 hour. After washing the secondary antibody three times or more, 100ml each of the TMB solution (Sigma, USA) was added and reacted at room temperature for 10-30 minutes, 1N H 2 SO 4 by 50ml each to terminate the reaction. Absorbance was measured at 450 nm using an ELISA reader (TECAN, USA). Protein expression inhibition rate (%) was calculated by the following equation (4).
  • Examples 1 to 23 can act as an MC1R antagonist to show whitening efficacy, and at the same time inhibit the inflammation-related signaling processes below the ETbR (Table 13, 14), it can be seen that it has anti-inflammatory efficacy.
  • the cosmetic composition for skin whitening of the present invention not only provides an excellent whitening effect even in a small amount, but also shows a superior whitening effect even when compared to DermaPep TM UL, which is a conventional whitening raw material.

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Abstract

La présente invention concerne des dérivés dipeptidiques d'undécénoyle de formule chimique 1 suivante et une composition cosmétique de blanchiment de la peau en contenant. Les dérivés dipeptidiques d'undécénoyle de la présente invention agissent en tant qu'antagonistes des récepteurs de la mélanostimuline-α (α-MSH), les MC1R, afin d'inhiber la génération de mélanine dans les cellules. En outre, là où un antagoniste traditionnel des MC1R entraîne un blanchiment de la peau et provoque une réaction inflammatoire, lesdits dérivés dipeptidiques d'undécénoyle ne provoquent pas de réaction inflammatoire et il a même été démontré qu'ils ont pour effet d'inhiber la réaction inflammatoire provoquée par les UV. En outre, la stimulation dont les endothélines sont à l'origine a également été inhibée, du fait d'une réaction croisée au niveau du processus de signalisation aval des MC1R et des récepteurs à l'endothéline de type B. Lesdits dérivés dipeptidiques d'undécénoyle ne sont pas toxiques pour l'organisme humain et pénètrent de façon remarquable dans la peau, présentent une très bonne stabilité et peuvent donc être très efficacement utilisés en tant que principes actifs cosmétiques de blanchiment de la peau, de même que pour leur effet anti-inflammatoire. [Formule chimique 1] CH2=CH(CH2)8-C(=O)-A1-A2-OR Dans ladite formule chimique, -C(=O)- désigne un groupe carbonyle, A1 et A2 représentent des résidus d'acides aminés et sont choisis, respectivement, parmi la Leu, la Phe, la Trp et la Gly et, plus précisément, parmi les formes lévogyre, dextrogyre ou mixte, de la Leu, de la Phe et de la Trp, R représente H, un groupe alkyle en C1-C18, un groupe phényle ou un groupe benzyle, un groupe undécénoyle se lie au groupe amine de A1, et OR se lie au groupe carbonyle de A2.
PCT/KR2012/000697 2011-03-18 2012-01-30 Dérivés dipeptidiques d'undécénoyle et composition cosmétique de blanchiment de la peau en contenant Ceased WO2012128464A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117752580A (zh) * 2023-12-22 2024-03-26 广州市腾宇化妆品有限公司 一种含有多肽的美白组合物及其应用
EP4247165A4 (fr) * 2020-11-17 2025-01-22 Hofseth Biocare ASA Traitements respiratoires

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5401773A (en) * 1990-02-22 1995-03-28 Roussel-Uclaf Lactic acid acylates
JPH0853332A (ja) * 1994-08-11 1996-02-27 Kao Corp 美白化粧料
JPH107540A (ja) * 1996-06-17 1998-01-13 Kao Corp 美白化粧料
US6242644B1 (en) * 1998-12-14 2001-06-05 Hoffmann-La Roche Inc. N-(4-carbamimidoyl-phenyl)-glycine derivatives
US20070232698A1 (en) * 2004-05-31 2007-10-04 Akira Shibuya Slimming Skin External Preparation and Cosmetic Containing the Same
US20080234224A1 (en) * 2004-03-31 2008-09-25 Motoaki Kamachi External Preparation For Skin

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101109737B1 (ko) * 2009-10-09 2012-02-15 미원상사주식회사 지방산 아미노에스테르 및 이를 함유하는 피부미백용 화장료

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5401773A (en) * 1990-02-22 1995-03-28 Roussel-Uclaf Lactic acid acylates
JPH0853332A (ja) * 1994-08-11 1996-02-27 Kao Corp 美白化粧料
JPH107540A (ja) * 1996-06-17 1998-01-13 Kao Corp 美白化粧料
US6242644B1 (en) * 1998-12-14 2001-06-05 Hoffmann-La Roche Inc. N-(4-carbamimidoyl-phenyl)-glycine derivatives
US20080234224A1 (en) * 2004-03-31 2008-09-25 Motoaki Kamachi External Preparation For Skin
US20070232698A1 (en) * 2004-05-31 2007-10-04 Akira Shibuya Slimming Skin External Preparation and Cosmetic Containing the Same

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4247165A4 (fr) * 2020-11-17 2025-01-22 Hofseth Biocare ASA Traitements respiratoires
CN117752580A (zh) * 2023-12-22 2024-03-26 广州市腾宇化妆品有限公司 一种含有多肽的美白组合物及其应用

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